Victor Viau


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Graduate Student Supervision

Doctoral Student Supervision

Dissertations completed in 2010 or later are listed below. Please note that there is a 6-12 month delay to add the latest dissertations.

Endocrine Adaptation to Repeated Psychological Stress in the Adult Male Rat Depends on Central Activation of Vasopressin V1a Receptors (2012)

The aim of this thesis was to investigate the brain pathways regulating the decline, or habituation of glucocorticoid responses to repeated exposure of psychological stress. In the first study, adult male rats were exposed to repeated bouts of restraint stress and tested for changes in endocrine responses and neuropeptide expression in the brain. Stress-induced elevations in glucocorticoids decreased as restraint was repeated, and this decline was associated with increased mRNA levels of the neuropeptide vasopressin in the medial amygdala and the bed nucleus of the stria terminalis. This suggested that central vasopressin signaling might be involved in coordinating glucocorticoid habituation. In a follow up experiment, adult male rats were again repeatedly restrained, however this time animals received continuous intracerebroventricular infusion of saline or an antagonist for the vasopressin V1a receptor. As expected, saline treated males showed stress-induced increases in vasopressin mRNA levels and habituation of glucocorticoid responses. Blocking activation of the vasopressin V1a receptor had no effect on basal or acute stress glucocorticoid levels. Antagonism did however attenuate habituation of glucocorticoid responses during subsequent restraint exposures. These findings indicated that habituation of endocrine stress responses depends on central vasopressin signaling. In the final set of experiments, saline and antagonist treated male rats were again repeatedly restrained, but this time brains were analyzed for changes in cellular activation using immunohistochemical detection of Fos protein. Antagonism had no effect on basal Fos expression, but blunted the habituation of repeated stress-induced Fos responses within the hypothalamus and select forebrain regions. Furthermore, a separate experiment revealed that repeated restraint induces region-specific changes in central V1a receptor binding levels which included decreases in the hippocampus, thalamus and central amygdala, but increases in the septum and bed nucleus of the stria terminalis. Overall, these studies show that habituation of glucocorticoid responses depends on central vasopressin V1a receptor signaling. The findings also suggest that stress-induced increases in V1a receptor binding within the bed nucleus and septum might be especially relevant in promoting habituation of glucocorticoid responses.

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Role of Neonatal Testosterone in Shaping the Adult Male Hypolthalamic-Pituitary-Adrenal Axis Response to Stress (2012)

Testosterone exposure during critical periods of development exerts major organizing effects on the hypothalamic-pituitary-adrenal (HPA) axis. The aim of this thesis was to determine how HPA axis activity is altered by neonatal testosterone, and where and how this might occur in the adult brain. In chapter two, I demonstrate that neonatal gonadectomy increased plasma corticosterone and Fos activation in the ACTH-regulating zone of the paraventricular nucleus of the hypothalamus under basal conditions and following restraint exposure. These responses were normalized with postnatal, but not adult testosterone replacement. Neonatal gonadectomized rats also had decreased numbers of AR and arginine vasopressin-positive cells in the bed nucleus of the stria terminalis and medial amygdala. This suggests that testosterone exposure during the neonatal period may prime adult HPA response to testosterone by altering AR levels and function within afferent mediators of HPA axis activity. Testosterone in the brain can be converted to estradiol by the aromatase enzyme, and estradiol usually impacts brain development directly. In chapter 3, I demonstrate that animals neonatally exposed to aromatase, or AR blockade fail to show a normal decline in corticosterone, or habituate, in response to repeat restraint exposure. By contrast, males castrated as adults show a significant reduction in corticosterone after repeated stress. These findings suggest an organizing influence of both ARs and estrogen conversion on HPA habituation, which occurs independently of activational effects of testosterone. The immediate early gene c-fos is rapidly induced in many brain regions following acute restraint stress and is an excellent tool for mapping functional differences in brain activation by stress. In chapter 4, we used c-fos mRNA as a tool to map changes in cellular activation in acutely stressed adult animals that received aromatase blockade neonatally. This treatment enhanced stress-induced c-fos expression in several limbic regions, including within the anterior cingulate and medial prefrontal cortex, lateral septum, anterior hypothalamic area, dorsal medial hypothalamus and medial amygdala, as well as at multiple levels of somatosensation. Based on these results, I propose estrogens exert effects during the neonatal period that result in systems-wide differences in adult neuroendocrine responses to homeostatic threat.

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Unmasking the functional anatomy of medial preoptic nucleus-influences on the hypothalmic-pituitary-adrenal axis (2009)

The hypothalamic-pituitary-adrenal (HPA) axis is a critical mediator of the stress responsesystem. However, despite clear evidence for an inhibitory role of testosterone on stress-inducedactivation of the HPA axis, the routes and mechanisms have not been addressed. To firstdetermine where testosterone acts in the brain to regulate stress-related input to the HPA axis, Iused a combined retrograde transport and immunohistochemical procedure to characterize theanatomical nature by which androgen targets in the brain communicate with the paraventricularnucleus (PVN) of the hypothalamus, the initial point of the neuronally mediated stress response.The findings suggest that androgens could act throughout the brain, and on a large assortment ofbrain regions that innervate the PVN. Among the brain regions identified, neurons of the medialpreoptic nucleus (MPN), highly express androgen receptors and project abundantly to the PVN,suggesting that the MPN stands out as a potential site of integration between testosterone and theHPA axis. To test the functional role of these cells, I tested whether lesions of the MPN alter theinhibitory effects of testosterone on the HPA axis. By selectively removing cells in the MPN,testosterone regulation of the PVN and HPA axis was eliminated. Together, these findingsdemonstrated that the integrity of the MPN is essential in maintaining the regulatory effects oftestosterone on the brain's response to stress. Finally, to clarify whether the MPN is the seat of,or an obligatory relay for the central effects of testosterone, I tested the effects of implanting theandrogen receptor antagonist hydroxyflutamide into the MPN, on the stress-induced activation ofthe PVN and HPA output. The differential effects of androgen exposure in the MPN on thebiosynthetic capacity and activational responses of the PVN and its extended circuitriessuggested that the MPN is capable of bridging converging limbic influences to the HPA axiswith changes in gonadal status.

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Master's Student Supervision

Theses completed in 2010 or later are listed below. Please note that there is a 6-12 month delay to add the latest theses.

Organizational effects of gonadal hormones on the hypothalamic-pituitary-adrenal axis and glucocorticoid receptor responses in male and female rats (2016)

Neonatal gonadal hormones during critical periods of development can irreversibly alter the adult hypothalamic-pituitary-adrenal (HPA) axis. The aims of this thesis were to investigate the role of neonatal gonadal hormones to 1. have direct organizational effects on the HPA axis or via indirect effects on corticosterone binding globulin that lead to compensatory changes in HPA output and 2. if changes in HPA output are met by changes in glucocorticoid receptor (GR) responses. To assesses these questions, the neonatal hormone milieu in Long Evan rats were manipulated in males by blocking the conversion of testosterone to estradiol with an aromatase blocker (ATD) and in females by administering testosterone propionate (TP). As adults, we assessed the influence of neonatal hormone manipulations on plasma corticosterone levels and GR activation in response to acute and repeated restraint exposure. GR responses were assessed using western blots to analyze GR translocation (nuclear/ nuclear and cytoplasm) and phosphorylation of GR at the serine 211 site (Ser211). We then assessed if changes in GR translocation and Ser211 followed restraint induced changes in total and estimated free CORT levels. Our results showed significant differences in corticosterone levels in neonatal ATD treated male and TP treated female rats compared to their same sex control groups under basal-naïve conditions and after restraint exposure, respectively. In both sexes, for GR translocation and Ser211, there was a main effect of restraint stress exposure, and overall significant positive correlations with CORT (total and estimated free) levels. GR translocation was lower in neonatal TP treated females, with no effect in Ser211. Differences between male neonatal ATD and Sham groups were observed compared to untouched controls in Ser211, indicating the effects of the neonatal ATD treatment appear to be due to effects of surgery and gonadal hormone milieu exposure. Adult gonadal hormone levels differed between female neonatal groups, which are likely due to organizational effects of the neonatal treatments. The current study demonstrates neonatal gonadal hormones have long lasting effects on adult corticosterone outputs and sequential GR responses.

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A modifed version of the defensive burying paradigm exposes group differences in behavior, vasopressin and c-fos expression in testosterone-replaced rats (2013)

Testosterone has been shown to have marked effects on different types of coping behaviors. The current study was done to test the capacity by which testosterone alters the amount of defensive burying in the defensive burying paradigm. Twenty high and twenty low testosterone-replaced (T), castrated rats were used to determine the effects of testosterone using a modified version of the defensive burying paradigm. Rats were exposed to either a remotely triggered mousetrap (snap), or neutral trap (yoke), creating four groups; high-T snap, high-T yoke, low-T snap and low-T yoke. Under habituation conditions, both high- and low-T replacement groups show a characteristic behavioral profile that indicated they habituated. As a function of testosterone replacement, animals showed significantly less burying but more rearing with high-T compared to low-T replacement (p = 0.004 and p = 0.02, respectively). Under aversive trap conditions, testosterone showed a general effect to decrease burying behavior. Testosterone affected rearing behavior that was highest within high-T groups and with direct exposure to the snap. There were limited neurochemical effects of testosterone other than a significant increase in vasopressin mRNA expression in high-T compared to low-T animals in both the medial amygdala and posterior bed nucleus of the stria terminalis. In addition, animals that were exposed to either test condition showed higher c-Fos activation in the septum compared to transport controls. These results show that high testosterone replacement can reduce signs of neophobia, alter the magnitude of adaptable behaviors and promote proactive exploration under aversive conditions.

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Local and distal origins of limbic related projections to the paraventricular nucleus of the hypothalamus (2013)

Cortico-limbic circuits are often activated in response to emotional stress to provide salient information about individual stimuli. This allows organisms to recruit adequate response mechanisms, one of which is the hypothalamic-pituitary-adrenal axis. Cellular groups directly adjacent to the paraventricular nucleus of the hypothalamus (PVN) are in position to integrate and relay cortico-limbic projections to the PVN directly. Using a neuroanatomical approach we discovered unique patterns and strengths of cortico-limbic projections to separate cell groups in the PVN surround. Further, in response to restraint stress, dual retrograde labeling with the cellular activation marker FOS suggests these PVN surround circuits are involved in adaptive responses to emotional stress. Of the PVN surround subregions, the zona incerta (ZI) showed particularly dense connectivity with the medial prefrontal cortex (mPFC), lateral septum, and paraventricular thalamus. Due to the strength of this connectivity we sought to confirm that it can act as a relay site to the PVN through targeted injections of anterograde tracer. ZI injections of anterograde tracer led to terminally labeled fibers in the PVN confirming the capacity to influence hypophysiotropic neurons of the PVN. Additionally, there is the potential for the ZI to share functional cross-talk with other PVN surround subregions. In our final experiment we directed injections of retrograde tracer into the PVN in concert with anterograde injections in the mPFC resulting in convergence of tracer labeling within the ZI. The abutted signals strongly indicate the PVN surround is a promising site for limbic influence on HPA.

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