\r\n\tThe outcome of cancer therapy with radiation has been improving over the years due to technological progress. However, due to the biological property of cancer, current radiotherapy has limitations. Therefore, in consideration of the dynamics of tumor cells caused by radiation irradiation, attempts are being made to overcome the current drawbacks and to improve radiotherapy. It is expected that carbon ion beams, hyperthermia, oxygen effect, blood flow control, etc. will be used in the future in order to improve the treatments. This book aims to introduce research results of various radioprotective agent development research and hypoxia sensitizers.
",isbn:null,printIsbn:"979-953-307-X-X",pdfIsbn:null,doi:null,price:0,priceEur:0,priceUsd:0,slug:null,numberOfPages:0,isOpenForSubmission:!1,hash:"c574888a21d8152f6b25191ea63af008",bookSignature:"Prof. Yeunhwa Gu and Dr. Jin Ho Song",publishedDate:null,coverURL:"https://cdn.intechopen.com/books/images_new/9388.jpg",keywords:"Oxygen Effective Ratio, Low LET, Therapeutic Ratio, Radioresistance, Radiation Sensitivity, PLD Recovery, Enzyme Target, Oxygen Effective Ratio, Radioprotective Agent, Dose Reduction Factor",numberOfDownloads:null,numberOfWosCitations:0,numberOfCrossrefCitations:0,numberOfDimensionsCitations:0,numberOfTotalCitations:0,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"September 3rd 2019",dateEndSecondStepPublish:"September 24th 2019",dateEndThirdStepPublish:"November 23rd 2019",dateEndFourthStepPublish:"February 11th 2020",dateEndFifthStepPublish:"April 11th 2020",remainingDaysToSecondStep:"2 years",secondStepPassed:!0,currentStepOfPublishingProcess:5,editedByType:null,kuFlag:!1,biosketch:null,coeditorOneBiosketch:null,coeditorTwoBiosketch:null,coeditorThreeBiosketch:null,coeditorFourBiosketch:null,coeditorFiveBiosketch:null,editors:[{id:"248214",title:null,name:"Yeun-Hwa",middleName:null,surname:"Gu",slug:"yeun-hwa-gu",fullName:"Yeun-Hwa Gu",profilePictureURL:"https://mts.intechopen.com/storage/users/248214/images/system/248214.jpg",biography:"Dr. Yeun-Hwa Gu is working as a Professor for the Department of Radiation Oncology, Graduate School of Health Science, Junshin Gakuen University, Japan. 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Introduction
While less is known about the role of the amygdala in reward‐related learning compared to its role in fear conditioning where detailed circuitry has been mapped out, research to date nonetheless points to a very interesting and important function for the amygdala and distinct roles for sub‐nuclei of this structure. In this review chapter, we will focus on rodent studies (using rats and mice) examining the role of the basolateral (BLA) or central (CeA) amygdala in the formation and expression of both Pavlovian and instrumental associations, the effects of changes in reward magnitude or value on responding, and to changes in reward contingencies to define the role of the amygdala and subnuclei of this structure in learning about reward and control of reward‐seeking behaviours.
2. Basolateral nucleus of the amygdala
2.1. Involvement of BLA in encoding reward expectation in Pavlovian tasks
Neuronal firing in the BLA is elevated in response to reward‐predictive CS’s, occurring prior to reward delivery. Such activity is thought to drive reward‐seeking behaviours. For example, in an odour discrimination paradigm, BLA firing differs following presentation of stimuli that predict a positive outcome (sucrose) versus a negative outcome (quinine), suggesting the BLA is involved in learning about expectancy for consequences of a response [1]. Importantly, this discriminative neural activity precedes reliable behavioural discrimination, suggesting that the neural activity may support learning and behavioural change and is consistent with a role for the BLA in encoding of information about a reward predictive CS. Data regarding a causal role for this activity in Pavlovian learning are, however, more mixed.
A number of studies have found no evidence of any effect of BLA lesions on the acquisition of a Pavlovian response, with lesioned rats demonstrating food cup approach in the presence of a CS+ similar to that of sham rats ([2–4] see also [5]). The acquisition of Pavlovian autoshaping responses was also similar to that of controls [6]. However, other studies indicate BLA lesions or inactivation impairs acquisition of Pavlovian associations. Lesions of the BLA reduce rats’ preference for a flavoured solution paired with sucrose compared to sham controls, with no effect on consumption of CS+ and CS− solutions when these were not paired with sucrose, suggesting involvement of the BLA in Pavlovian associations between the flavour of the liquid and the sucrose reward [7]. BLA lesions impair taste‐potentiated odour aversion, and infusions of the GABA‐A agonist muscimol into the BLA indicate the BLA mediates the acquisition, but not the expression of taste‐potentiated odour aversion ([8], see also Ref. [9]). Also, lesions of the BLA impair the acquisition, but not expression of magazine approach in a task where discrete cues signal the location of a sucrose reward [10]. BLA lesions impair second‐order conditioning [3, 11, 12]; but this deficit is secondary to the role of the BLA in the assignment of motivational value to the first‐order CS+ without which motivational significance cannot then be transferred to the second‐order CS+ to produce conditioned responding [12]. Inactivation of the BLA with the NMDA antagonists AP‐5 or d,l‐2‐2‐amino‐5‐phosphonovalerate (D‐APV) impairs acquisition, but not expression of Pavlovian conditioned approach for sucrose or taste potentiated odour aversion [13, 14], while inhibition of dopamine D1 receptor activity with SCH‐23390 impairs acquisition of Pavlovian discriminative stimulus responding (i.e. approaching the food cup in the presence of a CS+, but not a CS−) [15]. D1 antagonism has no effect on responding when animals are trained further and tested drug free, suggesting a specific role for BLA D1 receptors in the performance of Pavlovian discriminative stimulus approach [15]. It appears acquisition of Pavlovian associations rely on a BLA to nucleus accumbens (NAcc) pathway, as optogenetic inactivation of the BLA to NAcc pathway using halorhodopsin impairs acquisition of licking behaviour for sucrose in response to sucrose predictive cues ([16], see also Ref. [17] for related function of this pathway). When optical stimulation was removed, licking returned to non‐stimulation levels, suggesting no long lasting effects of BLA to NAcc inhibition on subsequent task acquisition [16]. Thus, despite some inconsistencies in the literature, it appears that the BLA is required for acquisition, but not for expression of Pavlovian stimulus‐reward associations.
There is strong evidence implicating the BLA in the acquisition and expression of conditioned place preference (CPP) for a food reward. Electrolytic and neurotoxic lesions of the lateral amygdala (LA) impair acquisition of CPP for a food reward [18], while BLA lesions or inactivation performed after acquisition of CPP for a food reward impair expression of CPP [19, 20], suggesting that BLA is implicated in both the acquisition and expression of Pavlovian place learning. Furthermore, muscarinic receptors in the BLA are required for the consolidation of food CPP, as intra‐BLA scopolamine infusions following conditioning sessions impairs consolidation of food CPP [21]. Disconnection of the BLA from the NAcc also impairs expression of sucrose CPP, implicating this pathway in the expression of context‐food associations [19]. One study has demonstrated no effect of BLA lesions on place conditioning [10]; however, in this study, discrete cues were used to signal the presence of reward within a y‐maze, providing an alternative strategy by which the animals could solve the task and as discussed above, there are a number of studies demonstrating no effect of BLA lesions on Pavlovian learning using a discrete cue. As such, the majority of research suggests the BLA, and its projections to the NAcc are required for the acquisition and expression of CPP.
2.2. Involvement of BLA in instrumental learning
The involvement of the BLA in the acquisition and expression of instrumental appetitive learning has been extensively examined, with somewhat mixed results. For example, infusion of the NMDA antagonist AP‐5 [22] or the D1 antagonist SCH23390 [23] into the BLA prior to training has been reported to impair acquisition of lever pressing for sucrose pellets but once learning has occurred, BLA inactivation via AP‐5 or SCH‐23390 has no effect on the expression of action‐outcome contingencies, suggesting involvement of the BLA in task acquisition, but not expression [22, 23]. It is important to note that in these studies, performance of the lever‐press result produced not only primary reward but also a range of visual cues, e.g. offset of the houselight and onset of a stimulus light followed by sucrose at a 3 second delay. The presence of these stimuli as well as the delay in reward delivery makes it unclear whether the BLA is involved in acquisition of the instrumental response‐outcome contingency or these other aspects of the task. Indeed, other studies using a more pure instrumental design where the instrumental response produces reward without the presence of any stimuli or secondary reinforcers report no effect of BLA lesions on acquisition of instrumental responding for a single action‐outcome contingency (e.g. lever press delivers food pellets) ([2, 24], but see [25]). Furthermore, lesions of the BLA do not impair acquisition of instrumental responding when two action‐outcome contingencies earning distinct outcomes are trained (e.g. lever press delivers sucrose solution, chain pull delivers food pellets) [4, 26, 27]. In more complex discriminative stimulus tasks, where rats are required to initiate the correct action following stimulus presentation, BLA lesions or inactivation using the combined GABA A (muscimol) and B (baclofen) agonists [28, 29] or selective serotonin lesions of the BLA [30] do not impair task acquisition, suggesting that the BLA is not essential for stimulus‐response learning despite its role in stimulus‐reward learning. In contrast, BLA inactivation with muscimol or baclofen impairs expression of previously trained discriminative stimulus responding, suggesting that the BLA may contribute to task expression in a discriminative stimulus task but that when the BLA is inactivated during acquisition, the rats can solve the task, perhaps by using a different strategy [17].
2.3. Involvement of BLA in detecting changes in reward‐predictive nature of an action
In accordance with a role for the BLA in predicting reward, the BLA is also involved in detecting changes between an action and a rewarding outcome. This has been demonstrated through contingency degradation paradigms, in which the association between an action and its expected outcome is reduced through non‐contingent presentations of the reward. Lesions of the BLA performed prior to behavioural training impair contingency degradation, under conditions of extinction (when there is no opportunity to update action‐outcome contingencies) and also under conditions of partial reinforcement [26]. BLA lesions also impair contingency degradation when lesions are performed after acquisition of action‐outcome contingencies, ruling out any potential learning impairment which could impact the contingency degradation [31]. These studies suggest BLA involvement in detecting changes in the association between an action and expected reward.
2.4. Involvement of BLA in updating the value of an outcome
The BLA appears critical for generating internal representations of a reinforcer to guide choice. For example, BLA neurons are sensitive to reward magnitude. Rats in an eight arm radial maze exhibit differential BLA firing to rewards of high and low magnitude [32]. Also, BLA activity is altered in response to changes in expected reward magnitude (i.e. upshift or downshift in reward magnitude) [33]. However, once this contingency is learnt, BLA activity decreases, suggesting involvement of the BLA in the acquisition, but not expression, of reward magnitude changes [33]. Noradrenaline (NOR) is released in the BLA following an increase in the number of sucrose pellets delivered in an instrumental task, suggesting NOR in the BLA contributes to signalling changes in reward value [34].
In addition to changes in magnitude, the value of rewarding outcomes can change based on changes in the animal’s motivational state. Involvement of the BLA in updating the value of an outcome is demonstrated through incentive‐learning tasks. In these tasks, the value of a reinforcer is updated based on changes in internal states (e.g. hunger, satiety) and this information used to control goal‐directed responding. In an incentive learning paradigm, the experience of food consumption in a deprived state subsequently drives responding for that food in a future state of deprivation [35, 36]. Importantly, without the opportunity for direct contact with the reward in a novel motivational state, instrumental responding for that reward is not altered even when animals experience motivational state changes at test, e.g., responding is not increased despite an increase in hunger until the animal experiences that food while hungry. Consolidation and reconsolidation of this form of learning are blocked by intra‐BLA infusions of the protein synthesis inhibitor anisomycin [37]. Infusions of the non‐selective opioid antagonist naloxone into the BLA also impair acquisition but not retrieval of incentive learning [36]. This effect appears dependent on μ‐opioid receptors, as infusion of a μ‐receptor antagonist blocks acquisition of positive incentive learning (i.e. under conditions which enhance reinforcer value), and infusion of a μ‐receptor agonist impairs negative incentive learning (i.e. under conditions which reduce reinforcer value) [38]. K and δ antagonists have no effect on positive or negative incentive learning, suggesting a specific role for μ‐receptors in the BLA in incentive motivational processes [38].
Related results have been found in outcome devaluation studies. In this task, hungry rats are, for example, trained to perform two distinct instrumental responses each earning a unique food outcome. Rats are then pre‐fed one of these outcomes to satiety prior to a choice test where the two responses are available but no outcomes are delivered. BLA lesions or inactivation impairs sensitivity to outcome devaluation [4, 6, 26, 31, 39, 40]. This effect is observed when lesions are conducted both prior to instrumental learning, and after acquisition of instrumental learning [26, 31], indicating any potential effects of lesions on action‐outcome learning do not contribute to the loss of sensitivity to outcome devaluation. Also, BLA inactivation prior to the pre‐feeding treatment, but not after pre‐feeding but before the lever test impairs outcome devaluation, suggesting the BLA updates reinforcer value to guide choice, but that once reinforcer value has been updated, the BLA is no longer needed ([41], see also Ref. [6]). Such results have been taken as evidence that the BLA associates the specific sensory features of stimuli with motivational significance and updates this association as needed. This information can then be used to guide choice in outcome devaluation and related paradigms [26].
There are some reports of BLA lesions having no effect on outcome devaluation; however, it appears these results are due to differences in experimental parameters. When rats are trained in a Pavlovian magazine approach paradigm with a single reinforcer and devalued using lithium chloride (LiCl), BLA lesions do not impair outcome devaluation [3, 42]. There are a number of variables which could contribute to this apparent discrepancy, namely the method of training (Pavlovian or instrumental), the number of reinforcers used (one or two reinforcers) and the method of devaluation (LiCl or specific satiety). Johnson and colleagues [39] assessed the contribution of the method of training and method of devaluation to establish how these factors may help understand what aspect of learning requires BLA involvement. All rats were lesioned following training, to isolate the effect of the lesion to devaluation. All rats were trained with two reinforcers. Four groups of rats were used: rats were either given Pavlovian or instrumental training, and devalued via LiCl or specific satiety, creating the following four groups: Pavlovian‐LiCl, Pavlovian‐specific satiety, instrumental‐LiCl and instrumental‐specific satiety. BLA lesions impaired outcome devaluation in all four groups. As the only variable which was not manipulated was the number of reinforcers trained (in this case, two), this led the authors to suggest the number of reinforcers used may mediate whether the BLA is required for outcome devaluation. Indeed, if the BLA encodes sensory representations of a stimulus and associations of these with value, then successful devaluation performance may depend on the ability to generate sufficiently detailed outcome representations so that performance specifically related to the currently devalued outcome, but not other possible outcomes, being specifically affected. Thus, in the case of two reinforcers, the BLA is required to generate this specific representation so that animals can then directly respond to the two reinforcers based on specific sensory features in order to guide action selection as the motivational significance is largely overlapping. However, when only one reinforcer is present, no discrimination between reinforcers nor any association of value with the sensory features of the outcome is required, which may leave outcome devaluation intact when the BLA is offline ([39], for discussion, see Refs. [31, 43]).
2.5. The BLA, reward prediction and stimulus influences on responding
Despite some inconsistencies in the effects of BLA lesions or inactivation in the acquisition of instrumental responding, incentive learning and outcome devaluation tasks suggest that the BLA is important for assigning motivational significance to outcomes based on their specific sensory features. There is strong evidence to suggest that the BLA is important for assigning reward value to actions and external stimuli more generally. Early reports demonstrate that in an eight arm radial maze, BLA neurons exhibit enhanced firing to an anticipated reward encounter, implicating BLA activity in predicting rewarding events [32]. In operant tasks requiring rats to nose poke for sucrose, BLA firing is enhanced during reward expectation, but decreases when animals no longer anticipate reward delivery following their actions under behavioural extinction [44]. Also, LA neurons respond to reward predictive cues, and activity in LA neurons is associated with task efficiency and accuracy, as well as increased synaptic strength [45]. Finally, neurons activated in response to a discriminative stimulus fire in the BLA prior to the NAcc, suggesting the BLA drives NAcc neuronal responses to reward predictive cues to promote reward‐seeking behaviour [17].
Following detection of reward‐predictive stimuli, it appears glutamate transients in the BLA are involved in initiating reward‐seeking action. Glutamate transients in the BLA are enhanced during a seeking‐taking chain task for sucrose pellets, and glutamate transients tend to precede lever responses on both the distal lever (i.e. lever responses which gave access to the proximal lever) and proximal lever (i.e. lever responses on which are rewarded with sucrose) [46]. Furthermore, in a simple instrumental task, BLA glutamate transients are more likely to be associated with initiating the pressing bout, than with reward or non‐reward earning lever presses [47]. These data suggest glutamate signalling is critical for driving actions which lead to reward.
Experiments using outcome devaluation indicate involvement of the BLA in encoding the sensory specific properties of reinforcers. Further support for this notion is derived from Pavlovian‐instrumental transfer (PIT) experiments, where presentation of a CS previously paired with a reinforcer drives instrumental responding for the same reinforcer, despite the CS and instrumental response having never been trained together before. Importantly, when rats are trained with two reinforcers (i.e. two CSs are paired with two distinct rewards and two instrumental responses earn those same two rewards), responding during PIT can be identified as ‘specific’, with increased instrumental responding on the lever that, in training, delivered the same outcome as that predicted by the stimulus. In contrast, ‘general’ PIT is an elevation in responding that does not rely on a common outcome in instrumental and Pavlovian training phases (for a discussion, see [4, 48] #321). When rats are trained with multiple rewards, BLA lesions impair specific, but not general PIT [4, 27], suggesting the BLA is required for distinct stimuli to direct instrumental responding. Furthermore, blocking AMPA, but not NMDA receptors in the BLA inhibits PIT [47], and BLA glutamate transient frequency correlates with instrumental responding during the CS, which was trained with the same outcome, but not the different outcome [47]. Importantly, BLA glutamate transient frequency is enhanced during initiation of lever pressing, suggesting BLA engagement following detection of reward‐predictive stimuli which initiates goal‐directed responding [47]. It appears the involvement of the BLA in PIT is dependent on the number of reinforcers (stimuli and responses) trained, because when rats are trained with only one reinforcer, BLA lesions have no effect on PIT [2, 49]. When two reinforcers are trained, specific sensory properties need to be utilised to permit discrimination; this requires the BLA and is impaired following BLA lesions or inhibition. However, when only one reinforcer is trained, it is not necessary to distinguish between reinforcers via their sensory properties to direct responding; thus, this behaviour does not require the BLA and is therefore unimpaired following BLA lesions.
Consistent with PIT studies, BLA lesions abolish outcome‐guided responding in an outcome‐specific reinstatement task, in which outcome presentation selectively increases performance of a response previously associated with the same, but not a different outcome as that which was just presented [31], supporting the involvement of the BLA in the representation of sensory‐specific properties of stimuli and integration of those stimulus properties with motivational significance to direct choice behaviour.
2.6. The BLA mediates risky and effortful decision making
Experiments using delay discounting paradigms indicate the BLA guides choice towards high effort/high reward options. When rats are required to choose between high effort/high reward versus low effort/low reward options in a T‐maze, BLA lesions reduce choice for the high effort/high reward option [50, 51]. Similarly, when a high reward choice requires a longer delay, as in delay discounting paradigms, BLA lesions also bias choice to smaller, immediate reward [52, 53]. There can be some recovery from bias towards low effort/low reward options, suggesting the BLA is involved in the acquisition of the value of a reward in an effortful task [51]. Disconnection of the BLA from the medial prefrontal cortex or anterior cingulate cortex (ACC) also biases choice to smaller, immediate rewards ([50, 52] but see Ref. [54]), consistent with a role for these structures in effort‐based decision making [55].
Paradigms involving risky decision making indicate the BLA also guides choice towards high risk/high reward options. BLA inactivation with baclofen or muscimol induces a risk‐averse pattern of choice and, in a similar paradigm, reduces high effort choice, irrespective of the delay to reward [56]. It is possible that the BLA directs responding toward risk when loss is involved, as rats with BLA lesions bias their behaviour away from risk when loss was a consequence of a high risk choice, but do not alter their behaviour when potential gains are available [57]. BLA lesions or inactivation does not alter choice when two rewards are equal, or there is no risk involved [56–58], suggesting a particular role for the BLA in biasing choice in the face of aversive consequences. It appears a BLA‐NAcc connection mediates BLA‐induced biasing of choice, as contralateral lesions of these structures biases choice toward a less risky option [54].
Some studies demonstrate BLA lesions enhance, rather than decrease risky decision making in a rodent gambling task [59], or when foot shock is used as punishment (instead of reward omission [58]). However, a recent study indicates individual differences may help explain these results. BLA inactivation can affect animals differently at an individual level—BLA inactivation increases effortful choice in rats which, at baseline, chose low effort/low reward options and BLA inactivation decreases effortful choice in rats which, at baseline, chose high effort/high reward options [60]. Furthermore, this biasing of choice appears dependent on BLA dopamine receptors. In risk‐averse rats, D1 agonist infusions into the BLA increase risky choice, whereas in risk‐prone rats, enhancing D1 activity reduces risky choice [61]. Also, infusions of the D2 agonist quinpirole reduce risky choice in risk‐prone rats [61]. It is possible dopamine receptors in the BLA mediate the interaction between costs and benefits in a task to generate subjective value which could differ between individuals or across experiments. Approaching the BLA as a mediator of decision making based on a cost/benefit analysis may explain why some studies report an increase in risky decision making following BLA inactivation—the effects of inactivation of this structure on behaviour may be dependent on task parameters which can bias decision making in a certain direction.
2.7. No consistent involvement of the BLA in reversal learning
Several studies have examined the role of the BLA in reversal learning; however, the results at present are inconsistent. One study demonstrates inactivation of the BLA with muscimol impairs reversal learning in an odour discrimination task [52]; however, another study demonstrates no effect of BLA lesions on reversal learning in a go/no‐go odour task [62]. Interestingly, in this study BLA lesions ameliorated impairments in reversal learning induced by orbitofrontal cortex lesions, suggesting projections between these two regions may control reversal learning [62]. In an operant nose‐poking discriminative stimulus task, BLA lesions have been shown to facilitate reversal learning, and limit the number of mistakes made following feedback on an incorrect trial [63]. However, in a similar nose‐poking discriminative stimulus paradigm, serotonin depletion in the BLA had no effect on reversal learning [30]. It appears the involvement of the BLA in reversal learning is not dependent on the task employed, as similar tasks (e.g. odour discrimination, operant nose poking) report inconsistent effects of BLA inactivation on reversal learning. Further research in this field is required to more conclusively determine the role of the BLA in reversal learning.
2.8. Involvement of the BLA in the appetitive extinction learning
A number of studies demonstrate that the BLA is critical for the acquisition of appetitive extinction learning. Excitotoxic lesions of the BLA enhance resistance to extinction learning when a magazine light and sucrose reinforcer are omitted, indicating BLA lesions impair extinction learning [28]. However, in this study, the use of excitotoxic lesions did not permit analysis of whether BLA lesions impair encoding, consolidation or retrieval of extinction learning [28]. Inactivation of the caudal BLA with bupivacaine (a sodium channel blocker) impairs acquisition, but not retrieval of extinction of instrumental responding, demonstrating BLA involvement of the acquisition of extinction learning [64]. In apparent contrast, intra‐BLA infusions of the NMDA partial agonist DCS, which should increase rather than decrease activation of BLA neurons, prior to extinction learning in an odour discrimination task has been reported to impair extinction and enhance responding at a retention session [65]. While a number of studies demonstrate DCS‐enhanced extinction learning (reviewed in Refs. [66, 67]), it appears the timing of DCS administration is critical in determining whether it enhances or impedes extinction learning (for a discussion, see Ref. [65]). Nonetheless, this study [65] demonstrates that NMDA receptors in the BLA contribute to extinction of appetitive learning. Finally, a subset of BLA neurons respond specifically during extinction of operant nose poking for sucrose; this subset does not respond during task acquisition and activity of these neurons is inversely correlated with responding during extinction [68]. These studies demonstrate a critical role for the BLA in detecting the absence of an expected reinforcer during instrumental appetitive extinction and are in agreement with the role of the BLA in detecting changes in reward value.
There is also some evidence to support a role for BLA signalling in the extinction of Pavlovian appetitive learning. For example, when rats are trained to lick for sucrose in the presence of a combined tone/light CS+, the firing of BLA neurons during extinction correlates strongly with extinction behaviour [69]. Furthermore, a subset of BLA neurons which responded during extinction also respond during reinstatement, suggesting the BLA is a site of plasticity mediating responding for motivationally significant stimuli [69]. These studies suggest that the BLA mediates aspects of Pavlovian appetitive extinction; however, further research in this field is required to determine the precise role of the BLA in appetitive extinction.
2.9. The BLA as part of a broader circuit involved in reward‐related learning
It is important to recognise that the BLA does not operate in isolation to control learning and performance. Below we highlight some example of interactions of the BLA with other structures. This section is not meant to comprehensive but to provide examples of how the BLA interacts with other brain areas. The BLA has dense projections to the posterior dorsomedial striatum (pDMS), insular cortex (IC) and NAcc [70], which, following detection of a change in reinforcer value in the BLA, mediate aspects of goal‐directed responding, such as knowledge of and engagement in action‐outcome contingencies. A BLA‐IC connection is required to encode and retrieve changes in reinforcer value [71]. BLA inactivation using the NR2B NMDA antagonist ifenprodil prior to specific satiety, but not prior to a choice test impairs outcome devaluation, suggesting BLA involvement in encoding changes in reinforcer value. However, ifenprodil infusions into IC prior to specific satiety or a choice test impair devaluation, suggesting that the IC mediates expression of devaluation. Finally, ifenprodil infused unilaterally into BLA prior to specific satiety and into the IC prior to a choice test blocks expression of devaluation, but ifenprodil infused into the IC prior to specific satiety and then into BLA prior to choice has no effect on expression of devaluation. This suggests the BLA updates and encodes information about reinforcer value during specific satiety, sending information to the IC prior to choice, and at test, the IC retrieves this information to guide choice between actions [71].
Also, connections between the BLA and posterior dorsomedial striatum (pDMS) are required to direct action‐outcome responding following a change in reinforcer value. The pDMS is critical for updating action‐outcome contingencies, as changes in response‐outcome associations are impaired following pDMS lesions [72, 73]. It appears the pDMS is required to retrieve action‐outcome associations following a change in reinforcer value, as unilateral lesions of the BLA coupled with inactivation of contralateral pDMS prior to a choice test impairs expression of outcome devaluation [73]. This suggests information from the BLA regarding the specific value of outcomes is transferred to the pDMS, which retrieves action‐outcome associations to guide instrumental performance [73].
Finally, a connection between the BLA and NAcc shell is necessary for action selection following reinforcer devaluation. Disconnection of the BLA and NAcc via contralateral excitotoxic lesions impairs outcome devaluation, without reducing overall responding [74]. It is possible the BLA conveys sensory‐specific outcome information and/or changes in reinforcer value to the NAcc, where it is used to direct outcome‐appropriate instrumental responding [74]. Previous reports demonstrate that NAcc shell lesions impair the ability for action‐outcome cues to bias action selection in Pavlovian‐instrumental transfer [75], supporting the NAcc shell being a limbic‐motor interface structure [76]. Thus, it appears sensory‐specific information from the BLA is used to drive action selection in the NAcc shell, which can direct actions through motor output structures such as the ventral pallidum and medial dorsal thalamus.
3. Central nucleus of the amygdala
3.1. The CeA in Pavlovian learning
The CeA is involved in conditioning with both appetitive and aversive reinforcement [77], and one proposed role for the CeA is determining the valence of reinforcing events. For example, c‐fos immunoreactivity is increased in the medial CeA following exposure to a CS+ signalling food delivery, compared to a CS which did not signal food delivery [78]. CeA c‐fos immunoreactivity is also increased following exposure to a CS+ signalling foot shock, particularly in ventral regions of the structure, suggesting sub‐regions of the CeA may detect the valence of a CS [78]. We focus here on data related to appetitive learning.
When a visual or auditory CS is paired with food, rodents can acquire distinctive behaviours to CS presentation; they may orient themselves to the CS, either by approaching or rearing to a light or startling in response to a tone (orienting responses) or approach the site of food delivery, usually a food cup or magazine (conditioned approach). There is considerable evidence to support a role for the CeA in conditioned orienting responses to a Pavlovian CS+, but not for conditioned approach. Additionally, CeA lesions do not impair second‐order Pavlovian conditioned approach [3]. Lesions of the CeA prior to training impair the acquisition of conditioned orienting responses (e.g. rearing to a light), but leave conditioned approach intact [79–81]. Similarly, inactivation of the CeA with the AMPA antagonist NBQX impairs acquisition of orienting responses [80]. CeA lesions or inactivation after Pavlovian training have no effect on the expression of Pavlovian orienting responses or food cup approach, suggesting a role for the CeA in the acquisition, rather than expression of orienting responses [80]. While some studies report no effect of CeA lesions on Pavlovian learning, these studies have only assessed conditioned approach behaviour ([2, 82] see also [7]), supporting a dissociation between conditioned approach and conditioned orienting responses in the CeA.
The CeA may be involved in conditioned approach behaviour when rats are trained to approach the magazine following the presentation of a CS+, but not a CS−, and a discrimination score is created which depicts their approach following one CS presentation over another CS. In Pavlovian approach paradigms, each CS+ is associated with a reinforcer, but not with the absence of reinforcement. CeA lesions or intra‐CeA inactivation of D1 or D3 receptors reduces conditioned approach behaviour [15, 83, 84]. If the CeA is involved in discriminating positive or negative reward value (discussed below), CeA inactivation may impair this discrimination, leading to a lower discrimination score. Supporting this interpretation, Andrzejewski and colleagues reported equal nose poking rates between the CS+ and CS−, rather than an abolition of nose poking [15], which would support lack of discrimination between the two CSs but not an inhibition of nose poking following CeA inactivation. It is also possible that these effects relate specifically to dopamine function within the CeA.
3.2. Circuitry mediating conditioned orienting responses
In rats injected with fluorogold, a retrograde tracer, into the substantia nigra pars compacta (SNc) there was a greater number of c‐fos positive/fluorogold positive cells in the CeA following food‐tone pairings than unpaired food and tone presentations, implicating this pathway in conditioning [85]. Furthermore, contralateral lesions to disconnect the CeA and SNc impair orienting responses but not food cup approach, compared to an ipsilateral lesion control group [85]. Considering that the CeA has a substantial projection to the SNc that provides dopaminergic innervation to the dorsolateral straitum (DLS) [86, 87], it is possible that a CeA‐SNc‐DLS pathway mediates orienting responses to Pavlovian food CS’s. Evidence for this comes from the demonstration that unilateral lesions of the CeA coupled with dopamine depletion in the DLS in the opposing hemisphere impairs conditioned orienting responses for food pellets, while leaving food cup approach behaviour intact [88]. Similar results were obtained when the DLS was reversibly inactivated with lidocaine [88]. Recovery of conditioned orienting responses occurred on drug free days in rats previously treated with intra‐DLS lidocaine, suggesting no long lasting effects of CeA‐DLS inactivation on acquisition of conditioned orienting [88]. Together, these results suggest orienting responses to a Pavlovian cue are mediated by indirect connections between the CeA and the DLS likely via the SNc.
3.3. The CeA in instrumental learning
The CeA does not appear to be critical for the acquisition of instrumental action‐outcome contingencies. Lesions of the CeA do not impair instrumental learning when there is a single action‐outcome contingency (i.e. one lever, one reinforcer) [2, 82] or two action‐outcome contingencies (i.e. two levers, two reinforcers) [4]. There is some evidence that the CeA is involved in updating action‐outcome contingencies. The omission of an expected reward at test enhances c‐fos immunoreactivity in the CeA, suggesting CeA detects the lack of reward [89]. Furthermore, fluorogold injections in the SNc demonstrate these c‐fos positive CeA cells project to the SNc [89], implicating a CeA‐SNc pathway in the detection of changes in reward contingencies. Lesions of the CeA produce a mild impairment in performance when an expected reward of small magnitude is omitted [90]; however, lesions of the entire CeA and BLA combined substantially reduce sensitivity to omission and so the specific contribution of the CeA is somewhat unclear [91].
CeA involvement in the detection of changes in reward value appears to depend on the paradigm used to assess this change. In studies using the outcome devaluation task, where an outcome is devalued either with selective satiety or LiCl‐induced sickness, CeA lesions do not impair behavioural sensitivity to changes in reward value [3, 82], indicating no role in this evaluative process and further substantiating intact action‐outcome learning necessary for performance in this task. Of interest, CeA lesions prevent loss of sensitivity to outcome devaluation that typically occurs with over‐training, suggesting a role for the CeA in habitual behaviour [82]. Similar effects are observed following disconnection of the CeA from the DLS, produced by contralateral lesions of these structures, suggesting that the CeA sends a reinforcement signal to the DLS to strengthen the stimulus‐response (S‐R) association that is thought to underlie habit learning [82].
While the CeA is not necessary for normal sensitivity to devaluation, it is involved in learning about changes in the magnitude of reward. When rats are trained to run in a straight alley maze task for a large food reward, a downward shift in the magnitude of the food reward increases the latency of intact rats to reach the smaller reward [92]. Post‐shift lidocaine infusions into the amygdala, which were mostly aimed at the CeA, reduce the latency to reach a smaller reward, suggesting reduced sensitivity to the change in reward magnitude [92]. Similarly, pre‐training CeA lesions slow learning about a downward shift in reward magnitude in a straight alley maze, supporting a role for the CeA in detecting reward magnitude changes [93, 94]. More recently, optogenetic stimulation of CeA with channelrhodopsin was shown to enhance lever pressing for a sucrose pellet when both the delivery and consumption of this pellet were paired with laser stimulation, compared to delivery of sucrose pellets alone, suggesting CeA stimulation may enhance the perceived magnitude of a reward [95]. Finally, the μ‐opioid agonist DAMGO administered within the CeA enhances sniffing and nibbling at a food cup or reward predictive lever, suggesting enhanced reward value attributed to these stimuli following CeA μ‐opioid stimulation [96]. Collectively, these studies implicate the CeA in processing changes in reward magnitude. Performance may be spared where tasks rely on discrimination and choice between rewards based their relative value and distinguished using specific sensory properties. Such tasks rely instead on the BLA as described above. Together these findings are consistent with the idea that whereas the BLA is responsible for assigning and updating the value of specific outcomes based on their sensory properties, the CeA is responsible for a less specific reinforcement signal, accounting for its role in both habit learning and adjusting performance following changes in reward magnitude [43, 82].
3.4. CeA involvement in stimulus influences on instrumental responding
The CeA plays a role in signalling the general motivational information carried by stimuli but consistent with the studies above, not in detailed representation of the specific features of distinct rewards or their representation. Evidence for this comes from Pavlovian‐instrumental transfer (PIT) tasks, which assess control of instrumental responding by Pavlovian cues, despite the two types of training being conducted separately. PIT occurs when presentations of a CS (previously paired with a US) drives instrumental responding which was previously trained to obtain the same US. The involvement of the CeA in PIT is dependent on the type of PIT being examined. When rats are trained on one instrumental action‐outcome contingency and undergo Pavlovian training which involves one CS‐US association, CeA lesions impair PIT [2, 48, 82], while intra‐CeA infusion of the μ‐opioid agonist DAMGO enhance PIT [97]. However, when rats are trained on two instrumental action‐outcome contingencies and two Pavlovian CS‐US pairings, lesions of the CeA have no effect the outcome‐specific PIT that is generated by this type of training [4]. Importantly, in an experimental design where a third excitatory CS+ is introduced in the Pavlovian training phase and paired with a third reward not earned by either instrumental response, CeA lesions impair responding to the third CS, but leave outcome‐specific PIT intact, suggesting the CeA is involved in general appetitive arousal rather than directing outcome‐specific responding [4]. This suggestion also accounts for the experiments described above which use only one action‐outcome contingency and one CS‐US pairing; when there is no choice between CS driven responses, a reduction in general appetitive motivation reduces responding in general. These findings suggest the CeA encodes a reinforcement signal which is devoid of specific details about an outcome.
3.5. Some involvement of the CeA extinction of appetitive learning
Several electrophysiological studies implicate the CeA in the extinction of appetitive learning. For example, Toyomitsu et al. [69] recorded from neurons in the BLA, LA and CeA during extinction of Pavlovian licking for sucrose reward. CeA firing during extinction correlated with extinction of licking; however, this correlation was not as strong as BLA firing [69]. Furthermore, while there were changes in the firing rate of CeA neurons between extinction and reinstatement, this was not as pronounced as in BLA neurons [69]. Calu et al. [98] recorded from the CeA during an over‐expectation task where initially multiple stimuli are trained as independent predictors of reward (e.g. individual stimuli such as a tone, light, etc., each predict a food pellet). The critical manipulation comes when two or more of these stimuli are then presented together, as a compound. Animals typically increase responding to such a compound indicating that they expect more reward based on the multiple predictors (e.g. since tone and light alone previously predicted one pellet, the two stimuli together should predict two pellets). However, if this compound is followed only by the original reward (one pellet) behavioural responding decreases across trials, as does activity of the CeA [98]. Together with data from extinction paradigms, this suggests that the CeA may signal reward reduction in general, not just reward omission. A recent study by Iordanova et al. [99] examined this possibility by exploring the role of the CeA in updating reward expectancies following a reduction in reward achieved either through extinction, where reward was omitted entirely, or through generating over‐expectation where, due to the presence of multiple predictors, a large reward is expected but not received. In both paradigms, the majority of recorded cells showed an increase in firing during the period where food reward was delivered and also during the preceding stimulus presentations reflecting reward expectancy. Neural firing to the extinction stimulus was reduced across trials compared to a control stimulus. When a combination of previously rewarded stimuli was introduced to generate over‐expectation, neural firing to this compound was increased relative to a control compound in early trials but then equivalent in later trials, presumably as animals came to expect the reduced reward that was received. A subpopulation of the reward‐responsive cells showed a reduction in firing to both the extinction and over expectation trials, suggesting a common role in signalling reduced reward expectation. Importantly, this change in neural activity preceded and predicted the decline in behavioural responding observed under both extinction and over‐expectation conditions. Because these conditions involved the delivery of different amounts of reward (no reward in extinction whereas reward was still delivered in over‐expectation, albeit less than initially expected based on the stimuli) the similar changes in neural activity are unlikely to reflect absolute reward magnitude, but rather may signal the reduction in reward expectancy. It is possible that this reduction in reward creates an aversive motivational state, in which case these findings could be consistent with a more general role for the CeA in emotional learning [99].
4. Conclusion
While less is known about the role of the amygdala in reward‐related learning compared to its role in fear conditioning where detailed circuitry has been mapped out, research to date nonetheless points to a very interesting and important function for the amygdala. For example, the basolateral amygdala is involved in associating sensory‐specific aspects of different outcomes with the rewarding effects of that outcome, a function critical for choice between alternatives and behavioural control more generally. Further, the amygdala appears to be involved in updating representations of value both when the value of the outcome is changed, for example, following devaluation, or when the relationship between predictors and outcome delivery is changed, as in extinction. Thus, the amygdala plays an important role in reward‐related learning. With the advent of tools such as optogenetics, researchers can now go on to explore how these functions are achieved within the complex circuitry of the amygdala and associated structures.
\n',keywords:"reward, Pavlovian conditioning, instrumental learning, reinforcement, basolateral amygdala, central amygdala",chapterPDFUrl:"https://cdn.intechopen.com/pdfs/54509.pdf",chapterXML:"https://mts.intechopen.com/source/xml/54509.xml",downloadPdfUrl:"/chapter/pdf-download/54509",previewPdfUrl:"/chapter/pdf-preview/54509",totalDownloads:1200,totalViews:439,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:1,dateSubmitted:"June 14th 2016",dateReviewed:"February 10th 2017",datePrePublished:null,datePublished:"July 5th 2017",dateFinished:"March 22nd 2017",readingETA:"0",abstract:"There has been substantial research into the role of the amygdala in fear conditioning and extinction of conditioned fear. The role of the amygdala in appetitive conditioning is relatively less explored. Here, we will review research into the role of the amygdala in reward‐related learning. Research to date suggests that the basolateral and central amygdala are responsible for learning about distinct aspects of a reinforcing event. For example, the basolateral amygdala is essential for distinguishing and choosing between specific rewards based on the specific‐sensory properties of those rewards as well as updating the relative value of specific rewarding events. In contrast, the central amygdala is involved in encoding reinforcement more generally and for regulating motivational influences on responding. We will also review what is known about the role of the amygdala in extinction of reward‐related behaviours and highlight areas for future research.",reviewType:"peer-reviewed",bibtexUrl:"/chapter/bibtex/54509",risUrl:"/chapter/ris/54509",book:{slug:"the-amygdala-where-emotions-shape-perception-learning-and-memories"},signatures:"Rose Chesworth and Laura Corbit",authors:[{id:"193670",title:"Dr.",name:"Laura",middleName:null,surname:"Corbit",fullName:"Laura Corbit",slug:"laura-corbit",email:"laura.corbit@sydney.edu.au",position:null,institution:null},{id:"194020",title:"Dr.",name:"Rose",middleName:null,surname:"Chesworth",fullName:"Rose Chesworth",slug:"rose-chesworth",email:"rose.chesworth@gmail.com",position:null,institution:null}],sections:[{id:"sec_1",title:"1. Introduction",level:"1"},{id:"sec_2",title:"2. Basolateral nucleus of the amygdala",level:"1"},{id:"sec_2_2",title:"2.1. Involvement of BLA in encoding reward expectation in Pavlovian tasks",level:"2"},{id:"sec_3_2",title:"2.2. Involvement of BLA in instrumental learning",level:"2"},{id:"sec_4_2",title:"2.3. Involvement of BLA in detecting changes in reward‐predictive nature of an action",level:"2"},{id:"sec_5_2",title:"2.4. Involvement of BLA in updating the value of an outcome",level:"2"},{id:"sec_6_2",title:"2.5. The BLA, reward prediction and stimulus influences on responding",level:"2"},{id:"sec_7_2",title:"2.6. The BLA mediates risky and effortful decision making",level:"2"},{id:"sec_8_2",title:"2.7. No consistent involvement of the BLA in reversal learning",level:"2"},{id:"sec_9_2",title:"2.8. Involvement of the BLA in the appetitive extinction learning",level:"2"},{id:"sec_10_2",title:"2.9. The BLA as part of a broader circuit involved in reward‐related learning",level:"2"},{id:"sec_12",title:"3. Central nucleus of the amygdala",level:"1"},{id:"sec_12_2",title:"3.1. The CeA in Pavlovian learning",level:"2"},{id:"sec_13_2",title:"3.2. Circuitry mediating conditioned orienting responses",level:"2"},{id:"sec_14_2",title:"3.3. The CeA in instrumental learning",level:"2"},{id:"sec_15_2",title:"3.4. CeA involvement in stimulus influences on instrumental responding",level:"2"},{id:"sec_16_2",title:"3.5. Some involvement of the CeA extinction of appetitive learning",level:"2"},{id:"sec_18",title:"4. Conclusion",level:"1"}],chapterReferences:[{id:"B1",body:'[Schoenbaum G, Chiba AA, Gallagher M. Orbitofrontal cortex and basolateral amygdala encode expected outcomes during learning. Nat Neurosci. 1998;1(2):155-9.]'},{id:"B2",body:'[Hall J, Parkinson JA, Connor TM, Dickinson A, Everitt BJ. Involvement of the central nucleus of the amygdala and nucleus accumbens core in mediating Pavlovian influences on instrumental behaviour. Eur J Neurosci. 2001;13(10):1984-92.]'},{id:"B3",body:'[Hatfield T, Han JS, Conley M, Gallagher M, Holland P. 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Different roles for orbitofrontal cortex and basolateral amygdala in a reinforcer devaluation task. J Neurosci. 2003;23(35):11078-84.]'},{id:"B43",body:'[Balleine BW, Killcross S. Parallel incentive processing: an integrated view of amygdala function. Trends Neurosci. 2006;29(5):272-9.]'},{id:"B44",body:'[Tye KM, Janak PH. Amygdala neurons differentially encode motivation and reinforcement. J Neurosci. 2007;27(15):3937-45.]'},{id:"B45",body:'[Tye KM, Stuber GD, de Ridder B, Bonci A, Janak PH. Rapid strengthening of thalamo‐amygdala synapses mediates cue‐reward learning. Nature. 2008;453(7199):1253-7.]'},{id:"B46",body:'[Wassum KM, Tolosa VM, Tseng TC, Balleine BW, Monbouquette HG, Maidment NT. Transient extracellular glutamate events in the basolateral amygdala track reward‐seeking actions. J Neurosci. 2012;32(8):2734-46.]'},{id:"B47",body:'[Malvaez M, Greenfield VY, Wang AS, Yorita AM, Feng L, Linker KE, et al. 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Orbitofrontal cortex and basolateral amygdala lesions result in suboptimal and dissociable reward choices on cue‐guided effort in rats. Behav Neurosci. 2011;125(3):350-9.]'},{id:"B52",body:'[Churchwell JC, Morris AM, Heurtelou NM, Kesner RP. Interactions between the prefrontal cortex and amygdala during delay discounting and reversal. Behav Neurosci. 2009;123(6):1185-96.]'},{id:"B53",body:'[Winstanley CA, Theobald DE, Cardinal RN, Robbins TW. Contrasting roles of basolateral amygdala and orbitofrontal cortex in impulsive choice. J Neurosci. 2004;24(20):4718-22.]'},{id:"B54",body:'[St Onge JR, Stopper CM, Zahm DS, Floresco SB. Separate prefrontal‐subcortical circuits mediate different components of risk‐based decision making. J Neurosci. 2012;32(8):2886-99.]'},{id:"B55",body:'[Bailey MR, Simpson EH, Balsam PD. Neural substrates underlying effort, time, and risk‐based decision making in motivated behavior. 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Dissociable contributions of anterior cingulate cortex and basolateral amygdala on a rodent cost/benefit decision‐making task of cognitive effort. Neuropsychopharmacology. 2014;39(7):1558-67.]'},{id:"B61",body:'[Larkin JD, Jenni NL, Floresco SB. Modulation of risk/reward decision making by dopaminergic transmission within the basolateral amygdala. Psychopharmacology (Berl). 2016;233(1):121-36.]'},{id:"B62",body:'[Stalnaker TA, Franz TM, Singh T, Schoenbaum G. Basolateral amygdala lesions abolish orbitofrontal‐dependent reversal impairments. Neuron. 2007;54(1):51-8.]'},{id:"B63",body:'[Izquierdo A, Darling C, Manos N, Pozos H, Kim C, Ostrander S, et al. Basolateral amygdala lesions facilitate reward choices after negative feedback in rats. J Neurosci. 2013;33(9):4105-9.]'},{id:"B64",body:'[McLaughlin RJ, Floresco SB. The role of different subregions of the basolateral amygdala in cue‐induced reinstatement and extinction of food‐seeking behavior. Neuroscience. 2007;146(4):1484-94.]'},{id:"B65",body:'[Portero‐Tresserra M, Marti‐Nicolovius M, Guillazo‐Blanch G, Boadas‐Vaello P, Vale‐Martinez A. D‐cycloserine in the basolateral amygdala prevents extinction and enhances reconsolidation of odor‐reward associative learning in rats. Neurobiol Learn Mem. 2013;100:1-11.]'},{id:"B66",body:'[Chesworth R, Corbit LH. Recent developments in the behavioural and pharmacological enhancement of extinction of drug‐seeking. Addict Biol. 2017;22(1):3-43.]'},{id:"B67",body:'[Myers KM, Carlezon WA, Jr. D‐cycloserine effects on extinction of conditioned responses to drug‐related cues. Biol Psychiatry. 2012;71(11):947-55.]'},{id:"B68",body:'[Tye KM, Cone JJ, Schairer WW, Janak PH. Amygdala neural encoding of the absence of reward during extinction. J Neurosci. 2010;30(1):116-25.]'},{id:"B69",body:'[Toyomitsu Y, Nishijo H, Uwano T, Kuratsu J, Ono T. Neuronal responses of the rat amygdala during extinction and reassociation learning in elementary and configural associative tasks. Eur J Neurosci. 2002;15(4):753-68.]'},{id:"B70",body:'[Kelley AE, Domesick VB, Nauta WJ. The amygdalostriatal projection in the rat—an anatomical study by anterograde and retrograde tracing methods. Neuroscience. 1982;7(3):615-30.]'},{id:"B71",body:'[Parkes SL, Balleine BW. Incentive memory: evidence the basolateral amygdala encodes and the insular cortex retrieves outcome values to guide choice between goal‐directed actions. J Neurosci. 2013;33(20):8753-63.]'},{id:"B72",body:'[Corbit LH, Janak PH. Posterior dorsomedial striatum is critical for both selective instrumental and Pavlovian reward learning. Eur J Neurosci. 2010;31(7):1312-21.]'},{id:"B73",body:'[Corbit LH, Leung BK, Balleine BW. The role of the amygdala‐striatal pathway in the acquisition and performance of goal‐directed instrumental actions. 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Differential involvement of the central amygdala in appetitive versus aversive learning. Learn Mem. 2006;13(2):192-200.]'},{id:"B79",body:'[Gallagher M, Graham PW, Holland PC. The amygdala central nucleus and appetitive Pavlovian conditioning: lesions impair one class of conditioned behavior. J Neurosci. 1990;10(6):1906-11.]'},{id:"B80",body:'[Groshek F, Kerfoot E, McKenna V, Polackwich AS, Gallagher M, Holland PC. Amygdala central nucleus function is necessary for learning, but not expression, of conditioned auditory orienting. Behav Neurosci. 2005;119(1):202-12.]'},{id:"B81",body:'[Olshavsky ME, Song BJ, Powell DJ, Jones CE, Monfils MH, Lee HJ. Updating appetitive memory during reconsolidation window: critical role of cue‐directed behavior and amygdala central nucleus. Front Behav Neurosci. 2013;7:186.]'},{id:"B82",body:'[Lingawi NW, Balleine BW. Amygdala central nucleus interacts with dorsolateral striatum to regulate the acquisition of habits. J Neurosci. 2012;32(3):1073-81.]'},{id:"B83",body:'[Hitchcott PK, Phillips GD. Double dissociation of the behavioural effects of R(+) 7‐OH‐DPAT infusions in the central and basolateral amygdala nuclei upon Pavlovian and instrumental conditioned appetitive behaviours. Psychopharmacology (Berl). 1998;140(4):458-69.]'},{id:"B84",body:'[Parkinson JA, Robbins TW, Everitt BJ. Dissociable roles of the central and basolateral amygdala in appetitive emotional learning. Eur J Neurosci. 2000;12(1):405-13.]'},{id:"B85",body:'[Lee HJ, Groshek F, Petrovich GD, Cantalini JP, Gallagher M, Holland PC. Role of amygdalo‐nigral circuitry in conditioning of a visual stimulus paired with food. J Neurosci. 2005;25(15):3881-8.]'},{id:"B86",body:'[Gonzales C, Chesselet MF. Amygdalonigral pathway: an anterograde study in the rat with Phaseolus vulgaris leucoagglutinin (PHA‐L). J Comp Neurol. 1990;297(2):182-200.]'},{id:"B87",body:'[Lerner TN, Shilyansky C, Davidson TJ, Evans KE, Beier KT, Zalocusky KA, et al. Intact‐brain analyses reveal distinct information carried by SNc dopamine subcircuits. Cell. 2015;162(3):635-47.]'},{id:"B88",body:'[Han JS, McMahan RW, Holland P, Gallagher M. The role of an amygdalo‐nigrostriatal pathway in associative learning. J Neurosci. 1997;17(10):3913-19.]'},{id:"B89",body:'[Lee HJ, Gallagher M, Holland PC. The central amygdala projection to the substantia nigra reflects prediction error information in appetitive conditioning. Learn Mem. 2010;17(10):531-8.]'},{id:"B90",body:'[Judice‐Daher DM, Tavares TF, Bueno JL. Involvement of the basolateral complex and central nucleus of amygdala in the omission effects of different magnitudes of reinforcement. Behav Brain Res. 2012;233(1):149-56.]'},{id:"B91",body:'[Tavares TF, Judice‐Daher DM, Bueno JL. Large neurotoxic amygdala lesion impairs reinforcement omission effects. Behav Brain Res. 2014;266:1-6.]'},{id:"B92",body:'[Salinas JA, Packard MG, McGaugh JL. Amygdala modulates memory for changes in reward magnitude: reversible post‐training inactivation with lidocaine attenuates the response to a reduction in reward. Behav Brain Res. 1993;59(1-2):153-9.]'},{id:"B93",body:'[Salinas JA, Parent MB, McGaugh JL. Ibotenic acid lesions of the amygdala basolateral complex or central nucleus differentially effect the response to reductions in reward. Brain Res. 1996;742(1-2):283-93.]'},{id:"B94",body:'[Salinas JA, White NM. Contributions of the hippocampus, amygdala, and dorsal striatum to the response elicited by reward reduction. Behav Neurosci. 1998;112(4):812-26.]'},{id:"B95",body:'[Robinson MJ, Warlow SM, Berridge KC. Optogenetic excitation of central amygdala amplifies and narrows incentive motivation to pursue one reward above another. J Neurosci. 2014;34(50):16567-80.]'},{id:"B96",body:'[Mahler SV, Berridge KC. Which cue to “want?” Central amygdala opioid activation enhances and focuses incentive salience on a prepotent reward cue. J Neurosci. 2009;29(20):6500-13.]'},{id:"B97",body:'[Mahler SV, Aston‐Jones GS. Fos activation of selective afferents to ventral tegmental area during cue‐induced reinstatement of cocaine seeking in rats. J Neurosci. 2012;32(38):13309-26.]'},{id:"B98",body:'[Calu DJ, Roesch MR, Haney RZ, Holland PC, Schoenbaum G. Neural correlates of variations in event processing during learning in central nucleus of amygdala. Neuron. 2010;68(5):991-1001.]'},{id:"B99",body:'[Iordanova MD, Deroche ML, Esber GR, Schoenbaum G. Neural correlates of two different types of extinction learning in the amygdala central nucleus. Nat Commun. 2016;7:12330.]'}],footnotes:[],contributors:[{corresp:null,contributorFullName:"Rose Chesworth",address:null,affiliation:'- School of Psychology, University of Sydney, Sydney, Australia
- School of Medicine, Western Sydney University, Sydney, Australia
'},{corresp:"yes",contributorFullName:"Laura Corbit",address:"laura.corbit@sydney.edu.au",affiliation:'- School of Psychology, University of Sydney, Sydney, Australia
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Introduction
Thyroid cancer is the most frequent endocrine neoplasia. The National Cancer Institute estimated that there would be 44,670 new cases of thyroid cancer (TC) with 1690 deaths in 2010, and with an overall estimate of 56,870 new cases by 2017, and its incidence has been increasing in recent decades. Compared with other adult cancers, TC tends to occur in younger people between the ages of 20 and 60. It is three times more common in women than men [1] and has the fastest rising incidence rates in women and the second fastest in men with an annual percentage change of approximately 5%, making TC the sixth most common cancer in women [2]. There are four main types of which papillary and follicular (PTC, FTC) types together account for >90% followed by medullary thyroid cancers (MTC) with 3–5% and anaplastic carcinomas (ATC) making up <3% [3]. Reasons for this trend have been attributed to improvement in imaging (ultrasound technology) that is allowing the identification of ever smaller thyroid nodules. However, with this gain in detection, determining which benign nodules (adenomas) will progress to cancer cannot be determined on the basis of histology alone, underscoring the need for genetic markers of early detection for TC. Recently, epigenetic alterations have been shown to play a role in the development and progression of thyroid cancer.
With the deepening of tumor research, it has been gradually found that epigenetics plays an important role in the occurrence and development of tumors. Mechanisms of epigenetics include, but are not limited to, DNA methylation (methylating of ciliary carbon at position 5), posttranslation modification of histone, chromatin remodeling (structural change), gene imprinting, RNA interference (noncoding RNA or gene silencing), etc. The epigenetic mechanisms of tumor cells have lost a fine regulation, and the breakdown of epigenetic patterns will lead to tumor phenotype expression. These mechanisms have been reviewed elsewhere, and here we will focus on DNA methylation in thyroid cancer.
DNA methylation is an important epigenetic change, which is persistent and hereditary. The methylation of promoter CpG can regulate gene expression and maintain chromosome integrity and DNA recombination. Based on 789 samples from the Gene Expression Omnibus (GEO) and the Cancer Genome Atlas (TCGA) databases, the five-CpG signature could provide a novel biomarker with useful applications in thyroid cancer (PTC, FTC, ATC, and MTC) diagnosis and the diagnostic score formula on the condition of DNA methylation data [4]. However, the methylation pattern is unstable and can be reversed by small molecules and endogenous enzymes, leading to dedifferentiation and tumor heterogeneity. Abnormal DNA methylation, including a decrease in the overall methylation level of the genome, was accompanied by hypermethylation in some gene promoter regions. The hypermethylation of tumor suppressor gene promoter can reduce its expression, while the hypomethylation of tumor suppressor gene promoter can increase its expression, leading to tumorigenesis [5, 6, 7, 8]. The following will describe the state of aberrant DNA methylation in different thyroid cancers.
2. DNA methylation in PTC
RET/PTC rearrangement and mutations in Ras and BRAF genes often occur in papillary thyroid carcinoma (PTC) [9]. In addition, many methylation of cancer suppressor genes are associated with BRAF gene mutations, such as Ras-association domain family 1A (RASSF1A), solute carrier family 5 member 8 gene (SLC5A8), retinoic acid receptor β2 (RARβ2), tissue inhibitor of metalloproteinase3 (TIMP3), phosphatase and tensin homolog deleted on chromosome ten (PTEN), metallothionein 1G (MT1G), ataxia-telangiectasia mutated (ATM), E-cadherin (ECAD), death-associated protein kinase (DAPK), multiple tumor suppressor 1 (MTS1 or P16), and mut-L homolog 1 (MLH1). Mutations of TSHR gene are not common in thyroid cancer, but high methylation and low expression of TSHR gene often occur. Recent studies have found that thyroid-specific genes (thyroid-stimulating hormone receptor and sodium/iodide symporter (TSHR and NIS), thyroid transcription factor-1 (TTF-1)) play an important role in occurrence and development of PTC. This part summarizes the related research on methylation genes in PTC in recent years (see Table 1).
Sample sources | Main method | Related gene | Methylation (high/low) | Related gene expression (high/low) | BRAF mutation (+/−) | Cell pathway | Function | References |
---|
Tumor suppressor genes |
Patient tissue, normal people tissue | Western blotting analyses, RT-QPCR, fluorescent analysis | RASSF1A | High | Low | + | MAPK | Stabilize the microtubules | [12, 13] |
SLC5A8 | High | Low | + | Sodium transporter | [17] |
RARβ2 | High | Low | + | Negative regulation of cell cycle | [20] |
TIMP3 | High | Low | + | Inhibitor of metalloproteinase | [23] |
Patient tissue, normal people tissue | Methylation-specific PCR, RT-QPCR | PTEN | High | Low | + | PI3K/Akt | Inhibit PI3K/Akt pathway | [26] |
MT1G | High | Low | + | Metallothionein | [28] |
ATM | High | Low | + | Regulate cell cycle | [29] |
Patient tissue, normal people tissue | Database query and retrospective medical chart, RT-PCR | ECAD | High | Low | + | Wnt/β-catenin | Mediate the adhesion of cells | [8] |
Thyroid-specific genes |
Patient tissue, normal people tissue | Western blotting analyses, RT-QPCR | NIS | High | Low | + | TSHR/cAMP | Sodium transporter | [36] |
TSHR | High | Low | + | Thyrotropin receptor | [37] |
TTF-1 | High | Low | + | Inhibit the pro-adipogenic response to pioglitazone | [40] |
Other potential genes |
Patient tissue, thyroid cancer cell line | Western blotting analyses, RT-QPCR, bisulfite sequencing, and methylation-specific PCR | RASSF10 | High | – | – | Others | Inhibit cell proliferation | [42] |
14-3-3σ | High | – | – | Cell cycle regulation | [45] |
RIZ1 | High | – | – | Epigenetic mechanisms | [44] |
DACT2 | High | – | – | Inhibiting Wnt signaling | [45] |
Table 1.
Aberrant methylated genes in PTC.
2.1 DNA methylation of cancer suppressor genes in PTC
2.1.1 Ras association domain family 1 (RASSF1A)
RASSF1A is a member of Ras superfamily, which is located at chromosome 3 (exactly on 3p21.3). Hypermethylation of CpG islands in the RASSF1A promoter region contributes to epigenetic inactivation. It is a tumor suppressor gene widely expressed in various normal organs but is often deleted in tumors. It is speculated that BRAFV600E gene mutation in PTC regulated the RASSF1A-MST1-FoxO3 signaling pathway, which led to RASSF1A hypomethylation and affected the malignant degree of thyroid cancer. It is found that the methylation rate of RASSF1A in thyroid cancer is 15–75% [10]. Studies indicated RASSF1A methylation differed in PTC compared with normal thyroid and was correlated with extracapsular invasion inversely. It suggested that RASSF1A has a potential role as a molecular marker for characterization of PTC histopathology [11, 12, 13, 14, 15]. It is shown that hypermethylation of RASSF1A promoter region is 20–32% in PTC. Hypermethylation of RASSF1A in PTC was related to the multifocal and extracapsular invasion of tumors [16].
2.1.2 Solute carrier gene family 5A, member 8 (SLC5A8)
SLC5A8 is a passive iodine transporter located in the parietal membrane of thyroid follicular cells. SLC5A8 is not regulated by thyrotropin in normal thyroid tissues but methylated in thyroid tumors. Hypermethylation often occurred in the first exon of CpG islands in SLC5A8, which results in gene silencing and restoring expression inhibiting cancer cell growth. It has been pointed out that SLC5A8 was an anti-oncogene of colon cancer [17]. SLC5A8 was also frequently hypermethylated in thyroid cancer. Its function is unclear nowadays, but its hypermethylation might play a key role in the occurrence of thyroid cancer [18]. Studies revealed that SLC5A8 gene was highly methylated in typical PTC (90%) and only 20% in other types of PTC. In addition, low expression of SLC5A8 was also associated with BRAF T1796A, suggesting that SLC5A8 methylation may be important in MAPK pathway [19].
2.1.3 Retinoic acid receptor beta2
RARβ2 is a type of nuclear receptor that is activated by both all-trans retinoic acid and 9-cis retinoic acid, which has been shown to function as a tumor suppressor gene in different types of human tumors. It has been found that RARβ2 expression was decreased or deleted in tumors. It meant that RARβ2 inactivation was related to tumorigenesis. In the treatment of metastasis and recurrence of thyroid cancer, retinoic acid therapy could restore the iodine uptake ability of metastasis and then improved the efficacy of 131-I radiotherapy. Researches indicated that the methylation rate of RARβ2 in thyroid cancer was 14%, higher than that in normal thyroid tissues (7%). RARβ2 gene methylation was associated with BRAF gene mutation in Wnt/beta catenin pathway [20]. Studies found there was RARβ2 gene hypermethylation in thyroid cancer cell lines. And after treatment with 5-azacytidine, RARβ2 expression was significantly increased, and the growth of tumors was inhibited, while the inhibition still existed after removing 5-azacytidine. RARβ2 gene methylation took part in tumorigenesis and development in PTC [21].
2.1.4 Tissue inhibitor of metalloproteinases-3 (TIMP3)
TIMP3 can bind to matrix metalloproteinases (MMPs), inhibiting the activity of MMPs effectively [20, 22]. Methylation of TIMP3 promoter has been demonstrated in many malignant tumors. It is often associated with growth, invasion, and lymph node metastasis of malignant tumors. It is pointed out that BRAF mutation caused low expression of TIMP3 in PTC, which could cause invasion and progression of tumors. It was found that 38% of TIMP3 are hypermethylated in PTC [23].
2.1.5 Phosphatase and tensin homolog deleted on chromosome 10 (PTEN)
The PTEN gene is located on chromosome 10 (especially on the region of 10q23), which could encode a specific phosphatidylinositol triphosphate 3 dephosphorylation and inhibit the activation of PI3K/Akt signaling pathway. PTEN expression alteration is crucial to the pathogenesis of cancer and other diseases. Low level of PTEN caused by homozygous deletions, frameshift, nonsense mutations or hypermethylation, or PTEN protein destability occurs frequently in various human cancers [24, 25]. It was shown that PTEN gene expression was low in thyroid cancer. One study found the methylation status of PTEN in FA, FTC, and PTC. The results indicated that PTEN methylation level was gradually increased in PTC (45.7%), FA (83.3%), and FTC (85.7%). PTEN methylation was related to mutations genes in PI3K/Akt signaling pathway, such as PIK3CA and Ras genes, suggesting that PTEN methylation and PI3K/Akt signaling pathway played an important role in the process of occurrence and development in PTC [26].
2.1.6 Metallothionein 1G (MT1G)
MT1G, a member of the metallothionein family, is a highly conserved cysteine-rich small molecule, which is mainly involved in metal-related transport. MT1G exists in normal cells; it can regulate and maintain intracellular metal ion balance, cell proliferation, and apoptosis. MT1G promoter methylation is associated with decreasing gene expression, but not complete abrogation. Studies have shown that MT1G gene has abnormal methylation in thyroid cancer, liver cancer, colon cancer, and prostate cancer. It is confirmed that restoring MT1G gene expression could inhibit tumors growth in vivo and in vitro, suggesting that MT1G gene has anticancer effect [27]. It is shown that MT1G gene was abnormally methylated in thyroid cancer (30.3% in malignant tumors and 18.8% in benign tumors). Its expression was significantly decreased, and that methylation of MT1G gene was associated with its low expression. Further studies suggested that restoring MT1G gene expression could inhibit the growth and infiltration of PTC and induced cell cycle inhibition and apoptosis. The mechanism may inhibit PI3K/AKT pathway. In addition, hypermethylation of MT1G was also associated with lymph node metastasis [28].
2.1.7 Ataxia-telangiectasia mutated
ATM belongs to the PI3/PI4 enzyme family. Although there is emerging evidence for a role of ATM in promoting tumorigenesis, ATM signaling provides a barrier to activated oncogenes and tumor progression, rather than promoting cancer early in tumorigenesis. ATM is ubiquitous in human and other higher animal tissues and cells, such as testicular tissue. And a study showed that ATM was hypermethylated in PTC (50%) and 0% in normal thyroid tissue [29].
2.1.8 E-cadherin
ECAD is a family of transmembrane glycoproteins responsible for calcium-dependent cell adhesion. It is the key structural components of adherens junctions. It is reported that ECAD may also act as a gene transcriptional regulator. Two main mechanisms including hypermethylation of the promoter and microRNA imbalance have been widely studied under the ECAD regulation in head and neck tumors. The methylation of ECAD promoter region was accounted for 39.3% in PTC, and ECAD expression decreased in the early stage of tumorigenesis. The experiment data showed that methylation level of ECAD in thyroid cancer increased to 56% (18/32) and 0% (0/27) in normal thyroid tissue. Further studies found that there was no significant correlation between ECAD methylation and T stage of lymph node metastasis in thyroid cancer. After 2.6 years of follow-up, the recurrence of thyroid cancer associated with ECAD methylation has no correlation [8].
2.1.9 Death-associated protein kinase
DAPK is a calmodulin-regulated ATK, which has an important role in the process of apoptosis. DAPK mechanism is largely due to promoter hypermethylation, leading to gene silencing. DAPK is ubiquitous in normal tissues. When the promoter of DAPK is methylated, it will cause an abnormal gene expression. Abnormal expression of DAPK can hinder the normal process of apoptosis and bring about tumorigenesis. And its low expression or deletion is one of the important mechanisms of cell carcinogenesis. It has been found that methylation of CpG island in the promoter region in DAPK is an important reason for expression silencing. The loss of DAPK protein in sporadic colorectal cancer is caused by the promoter hypermethylation. It existed in very small tumors. Therefore, the loss of DAPK gene plays an important role in the early stage of tumor formation. It is reported that DAPK promoter methylation accounts for 51% in PTC and abnormal methylation and DAPK gene silencing existed in many kinds of cancer cells [30]. It is pointed out that the high methylation level of DAPK gene was associated with tumor size and multiple lesions [31].
2.1.10 Multiple tumor suppressor 1 (MTS1 or P16)
MTS1 is hereafter called p16. It is an anti-oncogene in many tumors. The 5′-CpG fragment in the promoter region of P16 gene is the most susceptible to methylation, inhibiting its expression products. Abnormal expression of P16 gene can over-activate cyclin-dependent protein kinase 4 and stimulate abnormal cell proliferation, leading to tumorigenesis. Aberrant methylation of promoter region in P16 gene is the main cause for P16 gene inactivation [32]. Some scholars reported that the P16 gene was hypermethylated (35.9%) in PTC [33].
2.1.11 Mut-L homolog 1
MLH1 is one of the DNA mismatch repair genes located on the 3p21 region in chromosome 3. A correlation between MLH1 promoter methylation, specifically the ‘C’ region stops in MLH1 protein formation, can prevent the normal activation of DNA repair gene. Low expression of MLH1 gene was associated with BRAFV600E mutation and RET/PTC rearrangement. Hypermethylation of MLH1 promoter was found in colon cancer as well. A study indicated that abnormal methylation of MLH1 was significantly correlated with lymph node metastasis of PTC, suggesting that MLH1 might be a molecular marker of lymph node metastasis in PTC [34]. Another study found that there were abnormal methylation and low expression of MLH1 in thyroid cancer and MLH1 expression is associated with BRAF, IDH1, and NRAS gene mutations [35].
2.2 DNA methylation of thyroid-specific genes in PTC
2.2.1 Thyroid-stimulating hormone receptor and sodium/iodide symporter
On the other hand, methylation of thyroid-specific genes is also one of the causes for occurrence and development of PTC. Thyroid-specific genes mainly include TSHR, NIS, thyroglobulin (Tg), and thyroid peroxidase (TPO), which participate in thyroid iodine uptake and maintenance of normal thyroid function. Under normal conditions, TSH stimulates TSHR on thyroid follicular epithelial cells and activates NIS to ingest iodine into cells. TSH is produced by the pituitary thyrotrophs and stimulates thyroid functions using TSHR. The iodine ingested synthesizes thyroid hormones catalyzed by TPO and is stored in Tg. It is found that abnormal expression of these molecules is related to iodine metabolism in PTC. Studies have shown that the expression of TSHR, NIS, Tg, and TPO with BRAF mutated in thyroid cancer is decreased [36]. In PTC, both TSHR and NIS are abnormally methylated, and their expression is decreased. Low expression of TSHR and NIS may be related to the occurrence and development of tumors. It also reduces the uptake of iodine capacity in tumor cells. Scholars found TSHR and NIS become an important cause for PTC in 131-I radiotherapy [37, 38]. In human and rabbit thyroid cancer cells, BRAFV600E mutant, a carcinogenic homolog of murine sarcomatous virulent bacterium, could cause activation of BRAF/MEK/MAPK signaling pathway and expression silencing of thyroid-specific genes including TPO, Tg, TSHR, and NIS. At last, it resulted in the reduction of iodine uptake in PTC.
2.2.2 Thyroid transcription factor-1
TTF-1 is known as thyroid-specific enhancer-binding protein (T/EBP). It is a transcription factor with homologous domains in the thyroid, lung, and central nervous system. TTF-1 gene is located in region 14q13.3 on chromosome 14. It is comprised of three exons and two introns. Under physiological conditions, TTF-1 is stable positive in thyroid tissue. TTF-1 can regulate the expression of thyroid-related genes such as TG, TPO, TSHR, and NIS. Thus it acts a pivotal part in regulating growth, development, and function of thyroid. It showed hNIS mRNA expression loss might be related to methylation of thyroid-specific transcription factor genes. Abnormal methylation caused loss of transcription factor expression with indirect loss of hNIS mRNA expression through the KAT-5 and KAT-10 responses to 5-azacytidine treatment with acquisition of parallel TTF-1 and hNIS mRNA expression. It was found that insufficient expression of TTF-1 and Pax-8 may result in the decrease of activity of thyroglobulin gene promoter in thyroid cancer cells. Some confirmed that TTF-1 gene was expressed lowly in thyroid cancer [39]. Other researchers studied the methylation status of TTF-1 in thyroid cancer and found that TTF-1 gene was highly methylated and lowly expressed in thyroid cancer cell lines, but not in normal thyroid [40].
2.3 DNA methylation of oncogene gene in PTC
At present, BRAF gene mutation has been found in melanoma, ovarian serous tumor, colorectal cancer, glioma, liver cancer, and leukemia. A large number of studies also showed that BRAF gene mutation in PTC is closely related to methylation of tumor suppressor. BRAF gene is an important transducer for Ras/Raf/MEK/ERK/MAPK pathways. About 90% of T1799A point mutation in BRAFV600E happen in PTC, which can increase BRAF activity. The BRAF gene plays its biological role by activating MEK/ERK signaling pathway. The result showed that PTC with BRAF gene mutation had strong tissue invasiveness and was easy to infiltrate tissues around thyroid gland [41]. Methylation of TIMP3 gene, SLC5A8 gene, and DAPK gene, which are tumor suppressor genes, are related to BRAF gene mutation and PTC invasion. The overexpression of BRAFV600E gene could lead to silencing of some thyroid-specific genes (like NIS, TG, TPO) in the activation state of BRAF/MEK/MAPK pathway. It could cause iodine uptake activity decrease and ineffectiveness of radioiodine therapy. Therefore, it could affect thyroid cancer progression.
2.4 Other potentially aberrant methylation genes in PTC
In recent years, DNA methylation has become a new research hotspot. Many genes have been studied as potential abnormal methylation sites, including Ras association domain family 2 (RASSF2), Ras-association domain family 10 (RASSF10), disheveled-binding antagonist of beta-catenin 2 (DACT2), retinoblastoma protein-interacting zinc finger gene 1 (RIZ1), 14-3-3 protein family (14-3-3 sigma), and other signaling pathways such as JAK-STAT pathway, NF-kappa B pathway, HIF1 alpha pathway, and Notch pathway in thyroid cancer [42, 43, 44, 45]. However, there are relatively few reports on these genes and pathways in thyroid cancer. Most of the mechanisms are not clear at present and need further study.
3. DNA methylation in FTC
DNA methylation analysis revealed 2130 and 19 differentially methylated CpGs in PTC and follicular thyroid carcinoma (FTC), respectively [46]. Aberrant DNA methylation of tumor suppressor genes is common in FTC and ATC. Certain specific tumor suppressor genes are mainly PTEN, RASSF1A, Rap1-GTPase activating protein, and thyroid-specific gene TRSH in FTC. This part summarizes the related research on methylation genes in FTC in recent years (see Table 2).
References | Gene | Function | DNA methylation | BRAFE | Incidence% |
---|
Tumor suppressor genes |
[39, 40, 41, 42] | RASSFIA | RASSF1A localizes to microtubules and promotes their stabilization | ↑ | + | 75% of FTC |
[50] | RAP1GAP | RAP1GAPase-activating protein | ↑ | + | 38% of FTC |
[25] | PTEN | PTEN is involved in the regulation of cell cycle and preventing cells from growing and dividing rapidly | ↑ | + | 85.7% of FTC |
Thyroid-specific genes |
[58] | NIS | Sodium transporter | ↑ | + | 53.8% of thyroid cancers |
[58] | TSHR | Thyrotropin receptor | ↑ | + | 47% of FTC |
Oncogene genes |
[52, 53] | Maspin | A member of serine protease inhibitor | ↑ | + | 100% of WDTC |
Other genes |
[61] | DMCpG | — | ↑ | — | 84% of FTC |
[61] | RASAL1 | — | ↑ | — | 4.88% of FTC |
[57] | COL4A2 | — | ↑ | — | 56% of FTC |
[64] | RASSF10 | — | ↑ | — | 50% of FTC |
Table 2.
Aberrant methylated genes in FTC.
3.1 DNA methylation of tumor suppressor genes in FTC
3.1.1 PTEN
PTEN negatively regulates AKT/PKB signaling pathway. It is involved in regulation of cell cycle, cell growth inhibition, and rapid division [47]. Aberrant DNA methylation in this gene is also mostly reported in FTC. It confirmed that PTEN promoter hypermethylation was detected in six of seven (85%) FTC and five of six (83.3%) follicular adenomas. The results showed a high frequency of PTEN promoter hypermethylation, especially in follicular tumors. It means that it has a possible role in thyroid cancer [25]. Studies found methylation status of PTEN in FA and FTC. And PTEN methylation level was gradually increased in FA (83.3%) and FTC (85.7%). Authors have shown that methylation of PTEN promoter plays an important role in FTC [48].
3.1.2 RASSF1A
Differential expression of RASSF1A gene is related to occurrence of thyroid cancer. Aberrant DNA methylation is an important mechanism of RASSF1A gene inactivation. Studies found 44% benign adenomas, 75% follicular thyroid cancers tumors, and 20% PTC tumors harbored promoter methylation in greater than or equal to 25% of RASSF1A alleles by real-time quantitative methylation-specific PCR [39, 40, 41, 42]. Methylation frequency was higher in invasive thyroid cancer. It was found to be 70% of the RASSF1A methylation rate in FTC, 80% in MTC, and 78% in UTC compared with benign PTC [14]. Inactivation of RASSF1A in different stages of thyroid cancer was detected by tumor metastasis classification, and compared with FTC, only a small part of RASSF1A methylation in PTC is abnormal. These studies indicated follicular cell-derived thyroid tumorigenesis may be an early step [15, 49].
3.1.3 Rap1-GTPase-activating protein
RAP1GAP gene encodes a type of GTPase-activating protein that downregulates Ras-related protein activity. Ras oncoproteins are very important for both development and maintenance of many tumor types. RAP1GAP is involved in the regulation of mitosis and carcinogenesis in thyroid cells. Researchers aimed to determine the global patterns of aberrant DNA methylation in thyroid cancer using DNA methylation arrays [50]. And the study identified 262 and 352 hypermethylated and 13 and 21 hypomethylated genes in PTC and FTC, respectively. In addition, 86 and 131 hypermethylated genes were identified. Among these genes, four potential oncogenes (INSL4, DPPA2, TCL1B, and NOTCH4) were frequently regulated by aberrant methylation in primary thyroid tumors [51].
3.1.4 Mammary serine protease inhibitor (Maspin)
Besides, a member of the serine protease inhibitor superfamily named Maspin is a unique tumor suppressor gene encoding SERPINB5 gene. Epigenetic changes of Maspin expression occurred in the 5′ regulatory region of Maspin gene and involved cytosine methylation, histone deacetylation, and chromatin accessibility. The epigenetic deregulation frequently participates in tumorigenesis by inactivation of tumor suppressor genes. The association of promoter hypermethylation and gene silencing is an established oncogenic process in cancer. Promoter methylation of Maspin gene could lead to gene silencing in thyroid cancer, breast cancer, skin cancer, and colon cancer. Studies have indicated that overexpression of Maspin in gastric cancer, pancreatic cancer, and ovarian cancers resulted from CpG promoter of Maspin demethylation. A study detected DNA methylation status in Maspin promoter region, indicating that overexpression of the gene was the result of DNA hypomethylation [52]. It was closely related to the morphological dedifferentiation of thyroid cancer. Another study found 100% Maspin hypermethylation was closely associated with morphological dedifferentiation in thyroid cancers [52, 53].
3.2 DNA methylation of thyroid-specific genes in FTC
In addition to tumor suppressor genes and oncogenes, hypermethylation could lead to NIS and TSHR gene silencing in FTC, too. NIS methylation is of great significance in treatment of thyroid cancer. Therefore, abnormal methylation of these genes may be the pathogenesis or progression factor in FTC [54, 55]. Aberrant promoter methylation was examined in 24 tumor suppressor genes using methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) assay and methylation-specific PCR (MSP). In thyroid cancers, CASP8 (caspase-8), RASSF1, and NIS were methylated in 9/13, 10/13, and 7/13, respectively [56]. Some researches also found combination gene panels TPO and UCHL1 (ROC = 0.607, sensitivity 78%) discriminated FTC from FA and RASSF1 and TPO (ROC = 0.881, sensitivity 78%) discriminated FTC from normal. Methylation of TSHR distinguished PTC from FTC (ROC = 0.701, sensitivity 84%) and PTC from FA (ROC = 0.685, sensitivity 70%) [57]. And the six-gene panel of TIMP3, RARβ2, SERPINB5, RASSF1, TPO, and TSHR, which differentiates PTC from normal thyroid, had the best combination sensitivity (91%) and specificity (81%) of the panels addressing discrimination of cancer tissue by quantitative methylation-specific polymerase chain reaction (QMSP) in a retrospective cohort of 329 patients [58, 59].
3.3 Other abnormal methylation genes in FTC
Nowadays, there are a few studies on methylation of FTC genes. However, many genes still have been studied as potential abnormal methylation sites in FTC. For example, one of the ZIC families called ZIC1 (C2H2-type zinc finger proteins) is frequently hypermethylated in FTCs [60]. Moreover, 3564 differentially methylated CpGs (DMCpG) were detected in FTC and 84% hypermethylated with respect to normal controls. It is suggested that perturbed DNA methylation, in particular hypermethylation, is a component of the molecular mechanisms leading to FTC formation and that DNA methylation profiling might help in differentiating FTCs from their benign counterpart [61]. Also, others identified the presence of RASAL1 mutations, with a prevalence of 4.88% (n = 2 of 41) in FTC and 16.67% (n = 5 of 30) in ATC [62]. Studies found a more detailed analysis showing that 53.9% of the hypermethylated and 81.5% of the hypomethylated CpG sites identified in differentiated primary tumors (PTCs and FTCs) were also present in differentiated thyroid carcinoma-derived cancer cell [61]. Aside from that, COL4A2 was hypermethylated in 56% of the FTC samples by array measurement in the discovery series [57, 63]. Another study indicated that RASSF10 was frequently hypermethylated in thyroid cancer. It showed 50% of methylation frequency of RASSF10 in FTC and the highest (100%) in MTCs [64].
4. DNA methylation in ATC
Promoter methylation of PTEN is also common in anaplastic thyroid cancer (ATC) [65]. PTEN methylation is related to gene changes of PI3K Akt pathway in thyroid tumors, including PTEN mutations, various subtypes of Ras mutations, PIK3CA mutations, and amplification [66, 67]. One study analyzed 24 genetic alterations in the major genes of MAPK and PI3K-AKT pathways in 48 ATC samples and found that the majority of (81%) samples that harbored genetic alterations could be likely activated in both pathways [68]. Accordingly, another DNA methylation pan-cancer study focused on promoters found that thyroid carcinoma exhibited one of the lowest frequencies in both hypomethylation and hypermethylation events. And ATC exhibits a high frequency of DNA methylation alterations (tenfold higher than PTC) [69, 70]. A recent pan-cancer analysis on whole exome sequencing revealed that the mutation frequency in PTC was one of the lowest (approximately 1 change/Mb across the entire exome) among solid tumors, while the mutation frequency in ATC was at the opposite extreme and was closer to that in melanoma and lung cancer, exceeding 100 changes/Mb [71, 72].
In addition, solute carrier family 26, member 4 (SLC26A4) gene, encodes a transmembrane protein named pendrin with up to 15 predicted membrane spanning domains and affects the flow of iodine into follicular lumen. The following were reported: 71% of ATC, 44% of benign tumors, 46% of FTC, and 71% of PTC, with abnormal SLC26A4 gene methylation in 64 cases of primary thyroid tumors and 6 cases of thyroid tumor cell lines [18, 73]. In addition, 81.5% of hypermethylated genes and 89% of hypomethylated genes were also present in nondifferentiated primary tumors (MTCs and ATCs) and nondifferentiated thyroid carcinoma-derived cancer cell lines [74], while Ras protein activator like-1 gene (RASAL1) displayed MAPK- and PI3K-suppressing and thyroid tumor-suppressing activities, which can be impaired by the mutations. Hypermethylation and mutations of RASAL1 were found in 33.33% (n = 10 of 30) of ATCs and in 0 of 20 (0%) of benign thyroid tumors [62]. However, ATC showed more hypomethylation than hypermethylation events, indicating that hypomethylation is related to dedifferentiation [70]. The authors validated four genes (NOTCH4 and TCL1B in ATCs, INSL4 and DPPA2 in MTCs) that become aberrantly hypomethylated in nondifferentiated thyroid tumors. All of them have been proposed to have an oncogenic role in cancer. And NOTCH4 (a member of the Notch family of transmembrane receptors) is frequently overexpressed in thyroid tumors [75, 76].
This part summarizes the related research on methylation genes in ATC in recent years (see Table 3).
Authors | Gene | Function | DNA methylation | BRAFE | Incidence% |
---|
Oncogene genes |
[70] | TCL1B | An oncogene frequently activated by reciprocal translocations | ↑ | + | 64% of ATC |
[70] | NOTCH4 A | Member of notch family, which plays a role in a variety of developmental processes | ↑ | + | 45% of ATC |
Thyroid-specific genes |
[18, 73] | SLC26A4 | Dysfunctional pendrin | ↑ | — | 71% ATC |
Tumor suppressor genes |
[25] | PTEN | PI3K–AKT pathway | ↑ | — | 81% of ATC |
[62] | RASAL1 | MAPK- and PI3K-suppressing | ↑ | — | 33.33% of ATC |
Table 3.
Aberrant methylated genes in ATC.
5. DNA methylation in MTC
Medullary thyroid cancer is a neuroendocrine tumor originating from parafollicular C cells, and it is highly resistant to chemo- and radiotherapy [77]. Spry1 is a candidate tumor-suppressor gene in MTC. The hyper-activation of PI3K/Akt/mTOR cascade has a relevant role in the pathogenesis and progression of MTC. In fact, most of pro-oncogenic effects of RET and Ras mutations are modulated by the activation of PI3K/Akt/mTOR pathway [78, 79]. For patients with metastatic disease, standard treatment modalities include local treatments (radiofrequency ablation, radiation therapy, embolization) and systemic treatment chemotherapy and more recently tyrosine kinase inhibitors (TKIs) targeting RET protein. As experience has been limited to case reports or case series, response rates (RR) ranged from 0 to 25% for periods of up to a few months. In nondifferentiated thyroid tumors, INSL4 and DPPA2 become aberrantly hypomethylated, both of which have been proposed to have an oncogenic role in MTC.
5.1 Sprouty1 (Spry1)
Sprouty (Spry) family of genes is composed of four members in mammals (Spry1–4). The Spry1 promoter is frequently methylated in MTC, and that Spry1 expression is consequently decreased. These findings identify Spry1 as a candidate tumor-suppressor gene in MTC. In mammals, the situation is more complicated as Spry proteins have been shown to be activators or inhibitors of receptor tyrosine kinase signaling depending on the cellular context or the receptor tyrosine kinase analyzed [80, 81]. Spry family members have been proposed to function as tumor-suppressor genes in a growing list of cancerous malignancies, including prostate and hepatocellular carcinoma, B-cell lymphoma, or neuroblastoma [82]. Finally, the authors found that SPRY1 promoter is frequently methylated and its expression decreased in human MTC. The mRNA levels of Spry1 are detected in murine C cells of thyroid with real-time RT-PCR. In situ hybridization showed expression of Spry1 mRNA in the fourth pharyngeal pouch, where thyroid C-cells originate and beta-galactosidase staining of thyroids from 1-month-old Spry1LacZ/þ mice. One study used the hypomethylating agent 5′-aza-deoxycytidine (5′-Aza-dC) to confirm a causal relationship between promoter methylation and Spry1 expression [83]. As expected, TT cells treated with 5′-aza-dC showed an increase of approximately sixfold in the levels of Spry1 mRNA when compared to vehicle-treated cells. 5′-Aza-deoxycytidine (AZA), a demethylating agent, is in combination with the mTOR inhibitor everolimus in MTC cells (MZ-CRC-1 and TT). An innovative bioinformatic pipeline identified four potential molecular pathways implicated in the synergy between AZA and everolimus: PI3K-Akt signaling, the neurotrophin pathway, ECM-receptor interaction, and focal adhesion. Among these, the neurotrophin signaling pathway was most directly involved in apoptosis, through NGFR and Bax gene overexpression. Increased expression of genes involved in the NGFR-MAPK10-TP53-Bax/Bcl2 pathway during incubation with AZA plus everolimus was validated by western blotting in MZ-CRC-1 cells [84].
5.2 Insulin-like 4 (INSL4)
INSL4 (pro-EPIL) belongs to the insulin and insulin-like growth factor family and is expressed strongly during the first trimester of pregnancy by the differentiated syncytiotrophoblast [85]. It has been shown to be overexpressed in breast tumors with an aggressive phenotype [86], but the underlying mechanisms are still unknown. The aberrant overexpression of INSL4 in breast tumors, together with the aberrant promoter hypomethylation reported in this study, suggests that promoter demethylation might be a frequent mechanism of activation of INSL4 oncogene activation in cancer.
5.3 Developmental pluripotency-associated 2 (DPPA2)
DPPA2 is expressed early in the embryo’s development [87] but also in some tumor types [88]. Although the underlying molecular mechanism has not been reported yet, the authors’ data indicated that promoter hypomethylation might play an important role. The frequent promoter hypomethylation observed in nondifferentiated tumors might be relevant for treatment with demethylating drugs [51, 88].
6. Conclusion
More and more researches have realized that the occurrence of tumors is not only entirely determined by genes but also epigenetics. The changes of epigenetics in thyroid cancer are mainly manifested in the aberrant methylation of tumor suppressor genes and thyroid-related genes. Numerous studies on DNA methylation in thyroid cancer have improved our understanding of thyroid carcinogenesis. Some of the recent findings, including the huge catalog of DNA methylation alterations, the association of DNA hypomethylation with cancer progression and dedifferentiation, the existence of different methylomes related to different clinical and molecular phenotypes, and the influence of immune-infiltrating cells in tumor DNA methylation patterns, are most likely to lead the direction of future research in the field of DNA methylation in thyroid cancer. A large number of studies confirmed the importance of DNA methylation as a source of novel biomarkers for early diagnosis, therapeutic perspective, and prognosis evaluation in thyroid cancer. In addition, the design of specific target demethylation drugs, which reactivate the function of tumor suppressor genes, is expected to become a new scheme for cancer treatment. Therefore, further functional experiments in vitro and in vivo are necessary for better understanding of the meaning and potential mechanism of DNA methylation changes in thyroid cancer as well as the evaluation of candidate biomarkers through case-control studies and prospective trials.
\n',keywords:"thyroid carcinoma, DNA methylation, epigenetic inheritance, tumor suppressor genes, oncogene genes, thyroid-specific genes",chapterPDFUrl:"https://cdn.intechopen.com/pdfs/71481.pdf",chapterXML:"https://mts.intechopen.com/source/xml/71481.xml",downloadPdfUrl:"/chapter/pdf-download/71481",previewPdfUrl:"/chapter/pdf-preview/71481",totalDownloads:352,totalViews:0,totalCrossrefCites:0,dateSubmitted:"August 12th 2019",dateReviewed:"January 8th 2020",datePrePublished:"March 18th 2020",datePublished:"July 1st 2020",dateFinished:"March 18th 2020",readingETA:"0",abstract:"We have summarized increasing data from all kinds of experiment results of papers in recent years, which are associated with tumor suppressor genes, oncogenes, and thyroid-specific genes and attempt to elucidate the importance of epigenetic modifications and the mechanisms of aberrant DNA methylation in thyroid cancer in this review. The results showed that current articles have revealed the importance of epigenetic modifications and the different types of mechanisms in thyroid cancer. The mechanisms of DNA methylation related to thyroid cancer demonstrate that acquired epigenetic abnormalities together with genetic changes play an important role in alteration of gene expression patterns. Aberrant DNA methylation has been well known in the CpG regions. Among the genes identified, we have shown the status of DNA promoter methylation in papillary, follicular, medullary, and anaplastic thyroid cancer. It suggested that thyroid cancer subtypes present differential promoter methylation signatures, which will encourage potential thyroid cancer detection in its early stages, assessment of prognosis, and targeted cancer treatment.",reviewType:"peer-reviewed",bibtexUrl:"/chapter/bibtex/71481",risUrl:"/chapter/ris/71481",signatures:"Zhu Gaohong and Xie Lijun",book:{id:"8176",title:"DNA Methylation Mechanism",subtitle:null,fullTitle:"DNA Methylation Mechanism",slug:"dna-methylation-mechanism",publishedDate:"July 1st 2020",bookSignature:"Metin Budak and Mustafa Yıldız",coverURL:"https://cdn.intechopen.com/books/images_new/8176.jpg",licenceType:"CC BY 3.0",editedByType:"Edited by",editors:[{id:"226275",title:"Ph.D.",name:"Metin",middleName:null,surname:"Budak",slug:"metin-budak",fullName:"Metin Budak"}],productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"}},authors:null,sections:[{id:"sec_1",title:"1. Introduction",level:"1"},{id:"sec_2",title:"2. DNA methylation in PTC",level:"1"},{id:"sec_2_2",title:"2.1 DNA methylation of cancer suppressor genes in PTC",level:"2"},{id:"sec_2_3",title:"2.1.1 Ras association domain family 1 (RASSF1A)",level:"3"},{id:"sec_3_3",title:"2.1.2 Solute carrier gene family 5A, member 8 (SLC5A8)",level:"3"},{id:"sec_4_3",title:"2.1.3 Retinoic acid receptor beta2",level:"3"},{id:"sec_5_3",title:"2.1.4 Tissue inhibitor of metalloproteinases-3 (TIMP3)",level:"3"},{id:"sec_6_3",title:"2.1.5 Phosphatase and tensin homolog deleted on chromosome 10 (PTEN)",level:"3"},{id:"sec_7_3",title:"2.1.6 Metallothionein 1G (MT1G)",level:"3"},{id:"sec_8_3",title:"2.1.7 Ataxia-telangiectasia mutated",level:"3"},{id:"sec_9_3",title:"2.1.8 E-cadherin",level:"3"},{id:"sec_10_3",title:"2.1.9 Death-associated protein kinase",level:"3"},{id:"sec_11_3",title:"2.1.10 Multiple tumor suppressor 1 (MTS1 or P16)",level:"3"},{id:"sec_12_3",title:"2.1.11 Mut-L homolog 1",level:"3"},{id:"sec_14_2",title:"2.2 DNA methylation of thyroid-specific genes in PTC",level:"2"},{id:"sec_14_3",title:"2.2.1 Thyroid-stimulating hormone receptor and sodium/iodide symporter",level:"3"},{id:"sec_15_3",title:"2.2.2 Thyroid transcription factor-1",level:"3"},{id:"sec_17_2",title:"2.3 DNA methylation of oncogene gene in PTC",level:"2"},{id:"sec_18_2",title:"2.4 Other potentially aberrant methylation genes in PTC",level:"2"},{id:"sec_20",title:"3. DNA methylation in FTC",level:"1"},{id:"sec_20_2",title:"3.1 DNA methylation of tumor suppressor genes in FTC",level:"2"},{id:"sec_20_3",title:"3.1.1 PTEN",level:"3"},{id:"sec_21_3",title:"3.1.2 RASSF1A",level:"3"},{id:"sec_22_3",title:"3.1.3 Rap1-GTPase-activating protein",level:"3"},{id:"sec_23_3",title:"3.1.4 Mammary serine protease inhibitor (Maspin)",level:"3"},{id:"sec_25_2",title:"3.2 DNA methylation of thyroid-specific genes in FTC",level:"2"},{id:"sec_26_2",title:"3.3 Other abnormal methylation genes in FTC",level:"2"},{id:"sec_28",title:"4. DNA methylation in ATC",level:"1"},{id:"sec_29",title:"5. DNA methylation in MTC",level:"1"},{id:"sec_29_2",title:"5.1 Sprouty1 (Spry1)",level:"2"},{id:"sec_30_2",title:"5.2 Insulin-like 4 (INSL4)",level:"2"},{id:"sec_31_2",title:"5.3 Developmental pluripotency-associated 2 (DPPA2)",level:"2"},{id:"sec_33",title:"6. 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DOI: 10.1634/stemcells.2006-0269]'},{id:"B88",body:'[John T, Caballero OL, Svobodova SJ, Kong A, Chua R, Browning J, et al. ECSA/DPPA2 is an embryo-cancer antigen that is coexpressed with cancer-testis antigens in non-small cell lung cancer. Clinical Cancer Research. 2008;14(11):3291-3298. DOI: 10.1158/1078-0432.CCR-07-1322]'}],footnotes:[],contributors:[{corresp:"yes",contributorFullName:"Zhu Gaohong",address:"1026909611@qq.com",affiliation:'- Department of Nuclear Medicine, First Affiliated Hospital of Kunming Medical University, Kunming, China
'},{corresp:null,contributorFullName:"Xie Lijun",address:null,affiliation:'- Department of Nuclear Medicine, First Affiliated Hospital of Kunming Medical University, Kunming, China
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We carried out an experiment to induce traumatic occlusion in mouse PDL and analyzed the expression of HSPs. HSPs investigated acts differently depending on the time of expression. HSPs are constitutively expressed in the PDL and defend cells from stress and maintain homeostasis under normal conditions. During bone addition to the PDL on the tension side, HSP27 and HSP47, HSP70 also acts as molecular chaperone, which assists the maturation of bone morphogenetic proteins and aids osteoblast activation. In HSP 70 and HSP 47, mechanical stress is applied to the PDL on the tension side for a short period of time for alveolar bone repairing, and when abnormality occurs in the collagen structure fibroblasts of PDL, it functions at the injured site, whereby extracellular that promotes abnormal collagen secretion and stores the modified protein in the endoplasmic reticulum, there by controlling the decalcification of PDL. In other words, HSP47 and HSP70 are expressed in PDL fibroblasts on the pressure side damaged by application of mechanical stress and contribute to the repair of collagen tissue by activating PDL fibroblasts, supporting recovery from cell damage.",signatures:"Rina Muraoka, Keisuke Nakano, Hidetsugu Tsujigiwa, Hitoshi\nNagatsuka, Hirokazu Matsuda, Mihoko Tomida, Norimasa Okafuji,\nKazuhiro Yamada and Toshiyuki Kawakami",authors:[{id:"246203",title:"Dr.",name:"Toshiyuki",surname:"Kawakami",fullName:"Toshiyuki Kawakami",slug:"toshiyuki-kawakami",email:"kawakami@po.mdu.ac.jp"},{id:"246208",title:"Dr.",name:"Rina",surname:"Muraoka",fullName:"Rina Muraoka",slug:"rina-muraoka",email:"mura@po.mdu.ac.jp"},{id:"246209",title:"Prof.",name:"Mihoko",surname:"Tomida",fullName:"Mihoko Tomida",slug:"mihoko-tomida",email:"mtomi@po.mdu.ac.jp"},{id:"246212",title:"Prof.",name:"Norimasa",surname:"Okafuji",fullName:"Norimasa Okafuji",slug:"norimasa-okafuji",email:"okafuji@po.mdu.ac.jp"},{id:"246301",title:"Dr.",name:"Keisuke",surname:"Nakano",fullName:"Keisuke Nakano",slug:"keisuke-nakano",email:"keisuke1@okayama-u.ac.jp"},{id:"246302",title:"Prof.",name:"Hidetsugu",surname:"Tsujigiwa",fullName:"Hidetsugu Tsujigiwa",slug:"hidetsugu-tsujigiwa",email:"tsuji@dls.ous.ac.jp"},{id:"246303",title:"Prof.",name:"Hitoshi",surname:"Nagatsuka",fullName:"Hitoshi Nagatsuka",slug:"hitoshi-nagatsuka",email:"jin@md.okayama-u.ac.jp"},{id:"246304",title:"Dr.",name:"Hirokazu",surname:"Matsuda",fullName:"Hirokazu Matsuda",slug:"hirokazu-matsuda",email:"www.oqo34oiiis7.xxx@docomo.ne.jp"},{id:"246305",title:"Prof.",name:"Kazuhiro",surname:"Yamada",fullName:"Kazuhiro Yamada",slug:"kazuhiro-yamada",email:"yamadak@po.mdu.ac.jp"}],book:{title:"Periodontology and Dental Implantology",slug:"periodontology-and-dental-implantology",productType:{id:"1",title:"Edited Volume"}}},{title:"Bone Marrow Mesenchymal Cell Contribution in Maintenance of Periodontal Ligament Homeostasis",slug:"bone-marrow-mesenchymal-cell-contribution-in-maintenance-of-periodontal-ligament-homeostasis",abstract:"In general, remodeling phenomenon of the periodontal ligament (PDL) is occurring in all times. Thus, in the chapter, the word “maintenance” was used, and the chapter title is “Maintenance of Periodontal Ligament Homeostasis.” Our experimental data on the remodeling of the PDL with cell acceleration at the furcation area in this experimental model are recovered using the cells in situ and the bone marrow-derived cells (BMCs). BMC migration into the PDL tissues using green fluorescent protein (GFP) bone marrow-transplanted model mouse was examined. BMCs have abilities of cell migration and differentiation into tissues/organs in the body. The immunohistochemistry revealed that GFP-positive cells were detected in the PDL. GFP-positive cells were also positive to CD31, CD68, and Runx2 suggesting that fibroblasts differentiated into osteoclasts and tissue macrophages. In this way, Notch signaling involvement considered in our tentative examinations revealed that the experimentally induced periodontal polyp was examined; the cytological dynamics of the cells in granulation tissue are mainly from migration of undifferentiated mesenchymal cells of the bone marrow and differentiate into the tissue-specified cells. Furthermore, the data suggest that cell differentiation is due to Notch signaling.",signatures:"Toshiyuki Kawakami, Keiko Kaneko, Tatsuo Takaya, Saeka Aoki,\nRina Muraoka, Mihoko Tomida, Norimasa Okafuji, Masahito\nShoumura, Naoto Osuga, Keisuke Nakano, Hidetsugu Tsujigiwa and\nHitoshi Nagatuka",authors:[{id:"246203",title:"Dr.",name:"Toshiyuki",surname:"Kawakami",fullName:"Toshiyuki Kawakami",slug:"toshiyuki-kawakami",email:"kawakami@po.mdu.ac.jp"},{id:"246208",title:"Dr.",name:"Rina",surname:"Muraoka",fullName:"Rina Muraoka",slug:"rina-muraoka",email:"mura@po.mdu.ac.jp"},{id:"246209",title:"Prof.",name:"Mihoko",surname:"Tomida",fullName:"Mihoko Tomida",slug:"mihoko-tomida",email:"mtomi@po.mdu.ac.jp"},{id:"246212",title:"Prof.",name:"Norimasa",surname:"Okafuji",fullName:"Norimasa Okafuji",slug:"norimasa-okafuji",email:"okafuji@po.mdu.ac.jp"},{id:"246302",title:"Prof.",name:"Hidetsugu",surname:"Tsujigiwa",fullName:"Hidetsugu Tsujigiwa",slug:"hidetsugu-tsujigiwa",email:"tsuji@dls.ous.ac.jp"},{id:"246303",title:"Prof.",name:"Hitoshi",surname:"Nagatsuka",fullName:"Hitoshi Nagatsuka",slug:"hitoshi-nagatsuka",email:"jin@md.okayama-u.ac.jp"},{id:"246205",title:"Dr.",name:"Keiko",surname:"Kaneko",fullName:"Keiko Kaneko",slug:"keiko-kaneko",email:"info_omfbiol@po.mdu.ac.jp"},{id:"246206",title:"Prof.",name:"Tatsuo",surname:"Takaya",fullName:"Tatsuo Takaya",slug:"tatsuo-takaya",email:"tachy@po.mdu.ac.jp"},{id:"246207",title:"Prof.",name:"Saeka",surname:"Matsuda",fullName:"Saeka Matsuda",slug:"saeka-matsuda",email:"m_saeka@po.mdu.ac.jp"},{id:"246210",title:"Prof.",name:"Masahito",surname:"Shoumura",fullName:"Masahito Shoumura",slug:"masahito-shoumura",email:"shoumura@po.mdu.ac.jp"},{id:"246579",title:"Prof.",name:"Naoto",surname:"Osuga",fullName:"Naoto Osuga",slug:"naoto-osuga",email:"osuga@po.mdu.ac.jp"},{id:"246580",title:"Prof.",name:"Keisuke",surname:"Nakano",fullName:"Keisuke Nakano",slug:"keisuke-nakano",email:"pir19btp@okayama-u.ac.jp"}],book:{title:"Histology",slug:"histology",productType:{id:"1",title:"Edited Volume"}}}],collaborators:[{id:"49581",title:"Dr.",name:"Hiroyasu",surname:"Endo",slug:"hiroyasu-endo",fullName:"Hiroyasu Endo",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"179467",title:"Prof.",name:"Laurence",surname:"Walsh",slug:"laurence-walsh",fullName:"Laurence Walsh",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"University of Queensland",institutionURL:null,country:{name:"Australia"}}},{id:"205427",title:"Prof.",name:"Terry",surname:"Rees",slug:"terry-rees",fullName:"Terry Rees",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"205483",title:"Dr.",name:"Hideo",surname:"Niwa",slug:"hideo-niwa",fullName:"Hideo Niwa",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"205484",title:"Prof.",name:"Kayo",surname:"Kuyama",slug:"kayo-kuyama",fullName:"Kayo Kuyama",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"205485",title:"Dr.",name:"Morio",surname:"Iijima",slug:"morio-iijima",fullName:"Morio Iijima",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"205490",title:"Prof.",name:"Takanori",surname:"Ito",slug:"takanori-ito",fullName:"Takanori Ito",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"223300",title:"Dr.",name:"Maya",surname:"Oshima",slug:"maya-oshima",fullName:"Maya Oshima",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"223302",title:"Dr.",name:"Tae",surname:"Serizawa",slug:"tae-serizawa",fullName:"Tae Serizawa",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"235443",title:"Dr.",name:"Fardad",surname:"Shakibaie",slug:"fardad-shakibaie",fullName:"Fardad Shakibaie",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"University of Queensland",institutionURL:null,country:{name:"Australia"}}}]},generic:{page:{slug:"OA-publishing-fees",title:"Open Access Publishing Fees",intro:"The Open Access model is applied to all of our publications and is designed to eliminate subscriptions and pay-per-view fees. 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\n\nThe Open Access Publishing Fee (OAPF) is payable only after your full chapter, monograph or Compacts monograph is accepted for publication.
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\n\t- 4,000 GBP Compacts Monograph - Short Form
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I am also a member of the team in charge for the supervision of Ph.D. students in the fields of development of silicon based planar waveguide sensor devices, study of inelastic electron tunnelling in planar tunnelling nanostructures for sensing applications and development of organotellurium(IV) compounds for semiconductor applications. I am a specialist in data analysis techniques and nanosurface structure. 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Today his focus is on defining the growth and development strategy for the company.",institutionString:null,institution:{name:"TU Wien",country:{name:"Austria"}}},{id:"19816",title:"Prof.",name:"Alexander",middleName:null,surname:"Kokorin",slug:"alexander-kokorin",fullName:"Alexander Kokorin",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/19816/images/1607_n.jpg",biography:"Alexander I. Kokorin: born: 1947, Moscow; DSc., PhD; Principal Research Fellow (Research Professor) of Department of Kinetics and Catalysis, N. Semenov Institute of Chemical Physics, Russian Academy of Sciences, Moscow.\r\nArea of research interests: physical chemistry of complex-organized molecular and nanosized systems, including polymer-metal complexes; the surface of doped oxide semiconductors. He is an expert in structural, absorptive, catalytic and photocatalytic properties, in structural organization and dynamic features of ionic liquids, in magnetic interactions between paramagnetic centers. The author or co-author of 3 books, over 200 articles and reviews in scientific journals and books. He is an actual member of the International EPR/ESR Society, European Society on Quantum Solar Energy Conversion, Moscow House of Scientists, of the Board of Moscow Physical Society.",institutionString:null,institution:{name:"Semenov Institute of Chemical Physics",country:{name:"Russia"}}},{id:"62389",title:"PhD.",name:"Ali Demir",middleName:null,surname:"Sezer",slug:"ali-demir-sezer",fullName:"Ali Demir Sezer",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/62389/images/3413_n.jpg",biography:"Dr. Ali Demir Sezer has a Ph.D. from Pharmaceutical Biotechnology at the Faculty of Pharmacy, University of Marmara (Turkey). He is the member of many Pharmaceutical Associations and acts as a reviewer of scientific journals and European projects under different research areas such as: drug delivery systems, nanotechnology and pharmaceutical biotechnology. Dr. Sezer is the author of many scientific publications in peer-reviewed journals and poster communications. 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