Initiation of smoking mainly occurs during adolescence. Adolescents experience more stressful life events; therefore, stress may be a factor that contributes to this high risk of smoking initiation. The current study examines the effects of three different stressors (yohimbine, intermittent footshock and social defeat) on nicotine self-administration (NSA) in adolescent and adult rats. The effects of yohimbine and footshock were examined after the establishment of NSA behavior, while the effect of social defeat was tested on the initiation of NSA behavior. Yohimbine increased NSA, but the other two stressors did not. The increase in NSA induced by yohimbine tended to be higher in adults than in adolescents. No marked age differences in response to the other two stressors were observed. These results suggest that stress increases NSA in a stressor-specific manner, and that adolescents do not show enhanced vulnerability to the effect of stress on NSA.

Initiation of smoking mainly occurs during adolescence. Adolescents experience more stressful life events; therefore, stress may be a factor that contributes to this high risk of smoking initiation. The current study examines the effects of three different stressors (yohimbine, intermittent footshock and social defeat) on nicotine self-administration (NSA) in adolescent and adult rats. The effects of yohimbine and footshock were examined after the establishment of NSA behavior, while the effect of social defeat was tested on the initiation of NSA behavior. Yohimbine increased NSA, but the other two stressors did not. The increase in NSA induced by yohimbine tended to be higher in adults than in adolescents. No marked age differences in response to the other two stressors were observed. These results suggest that stress increases NSA in a stressor-specific manner, and that adolescents do not show enhanced vulnerability to the effect of stress on NSA.

Following social defeat, male hamsters exhibit behavioral changes characterized by a breakdown of normal territorial aggression and an increase in submissive/defensive behaviors in the presence of a non-aggressive intruder (NAI). We have termed this phenomenon conditioned defeat (CD). By contrast, only a small subset of defeated females exhibit submissive/defensive behavior in the presence of a NAI. We hypothesized that fluctuations in gonadal hormones might contribute to differences in the display of submissive behavior in intact female hamsters. Following social defeat, proestrous females (higher endogenous estradiol) were more likely to display conditioned defeat compared with diestrous 1 (lower endogenous estradiol) females. This finding suggests that there is an estrous cycle-dependent fluctuation in the display of CD in female hamsters and suggests that increased estradiol might contribute to increased submissive behavior. We then demonstrated that ovariectomized females given estradiol prior to CD testing exhibited significantly higher submissive behavior in the presence of a NAI suggesting that estradiol increases the expression of CD in female hamsters. We have also shown that castrated males that were singly housed for four weeks displayed significantly more submissive behavior than did their intact counterparts. Interestingly, castrated and intact males that were singly housed for 10 days prior to behavioral testing displayed similar behavior during CD testing. Together these data suggest that androgens and isolation modulate the display of CD in male hamsters. Finally, we examined brain activation following CD testing in defeated males and females (in diestrus 1 and proestrus). Defeated male and proestrous females exhibited increased Fos activation in the dorsal lateral septum and hypothalamic paraventricular nucleus relative to defeated diestrous 1 females. Diestrous 1 females exhibited increased Fos expression in the lateral bed nucleus of the stria terminalis compared with both defeated groups. Collectively, these data suggest that gonadal hormones and duration of individual housing modulate the display of CD in female and male hamsters and that those animals which display CD exhibit differences in patterns of neuronal activation than do those that do not display CD.

Social conflict is a part of everyday life, and it can be a potent stressor for both humans and other animals. In the laboratory, when two Syrian hamsters (Mesocricetus auratus) compete for territory, a dominance hierarchy is quickly formed. Becoming subordinate is a significant stressor resulting in increased release of adrenocorticotropic hormone, β-endorphin, and cortisol. Defeated hamsters will also subsequently fail to display territorial aggression in future social encounters and will instead display increased submissive behavior, even in the presence of a smaller, non-aggressive intruder. This change in behavior is consistent and long-lasting and has been termed conditioned defeat (CD). Corticotropin releasing factor (CRF) is an important neuropeptide in the control of the hypothalamo-pituitary-adrenal (HPA) axis response to stress. It is also involved in a number of behaviors such as anxiety, stress responding, food intake, learning, and memory. The widespread distribution of CRF, CRF-like peptides, and CRF receptors, particularly in brain sites related to anxiety, fear, and stress responses, suggests a role for CRF and CRF-like peptides in modulating emotional responses other than via HPA axis activity. It has also been shown that CRF may have a role in the acquisition and expression of CD. Non-specific and CRF type 2-specific CRF antagonists reduce the acquisition and expression of CD in male hamsters while injection of a CRF type 1-specific antagonist does not. Therefore, the goal of this dissertation was to investigate the role of CRF type 1 and 2 receptors in CD in hamsters and to identify neuroanatomical locations where CRF may be acting. It was found that non-specific or CRF type 1 receptor specific agonists enhance the expression, but not acquisition, of CD. Further, these agonists appear to enhance aggressive behavior in animals that were not previously defeated, suggesting a modulatory role for CRF type 1 receptors in agonistic behavior that depends on an animal’s previous social experience. Further, localization of CRF receptors was determined in hamster brain in sites thought important for CD and agonistic behavior, but changes in receptor binding following defeat were not observed. Implications of these results and future directions are discussed.

Social stress is a prevalent factor in society that can cause or exacerbate neuropsychiatric disorders including depression and posttraumatic stress disorder. According to the National Institutes of Health, 6.9% of adults in this country currently suffer from depression, and 4.1% suffer from an anxiety disorder. Unfortunately, current treatments are ineffective in reducing or alleviating symptoms in a majority of these patients. Thus, it is critical to understand how social stress changes in brain and behavior so that we might develop alternative treatments. Brain derived neurotrophic factor (BDNF), which binds to tyrosine kinase B (TrkB) receptors, plays a role in fear learning and in behavioral responses to stress, although we do not currently know whether BDNF promotes or prevents these responses. The purpose of this project was to understand how BDNF alters brain and behavior in response to social stress using a model of social stress in Syrian hamsters, termed conditioned defeat (CD). CD refers to the marked increase in submissive and defensive behavior following social defeat. Specific Aim (SA) 1 tested the hypothesis that BDNF, via TrkB receptors, promotes CD learning. Instead, we found that BDNF and a selective TrkB receptor agonist reduced CD and that a TrkB receptor antagonist enhanced CD. SA 2 tested the hypothesis that the behavioral response observed following systemic administration of TrkB-active drugs is mediated via their action in specific nodes of the neural circuit underlying CD. Unfortunately, the vehicle in which these drugs are dissolved independently activates immediate early gene expression making interpretation of these data impossible. Finally, SA 3 tested the hypothesis that BDNF alters defeat-induced neural activation at least in part by acting in the medial prefrontal cortex (mPFC). We demonstrated that BNDF microinjected into the mPFC site-specifically altered defeat-induced neural activation in the CD neural circuit supporting this hypothesis. Overall, these data suggest that BDNF acts to prevent social stress-induced changes in behavior, at least in part via the basolateral amygdala and the mPFC, and that BDNF-active drugs might be a useful avenue to pursue to discover new treatments for patients that suffer from stress-related neuropsychiatric disorders.

Thesis (Ph. D.)--Ohio State University, 2007. / Title from first page of PDF file. Includes bibliographical references (p. 75-89).

Submitted by Bruna Rodrigues (bruna92rodrigues@yahoo.com.br) on 2016-09-14T13:01:32Z No. of bitstreams: 1 TeseACC.pdf: 1221580 bytes, checksum: fd3199b489e81df5b5b589830f8ed420 (MD5) / Approved for entry into archive by Marina Freitas (marinapf@ufscar.br) on 2016-09-15T13:51:39Z (GMT) No. of bitstreams: 1 TeseACC.pdf: 1221580 bytes, checksum: fd3199b489e81df5b5b589830f8ed420 (MD5) / Approved for entry into archive by Marina Freitas (marinapf@ufscar.br) on 2016-09-15T13:51:55Z (GMT) No. of bitstreams: 1 TeseACC.pdf: 1221580 bytes, checksum: fd3199b489e81df5b5b589830f8ed420 (MD5) / Made available in DSpace on 2016-09-15T13:52:06Z (GMT). No. of bitstreams: 1 TeseACC.pdf: 1221580 bytes, checksum: fd3199b489e81df5b5b589830f8ed420 (MD5) Previous issue date: 2015-05-15 / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Stressful situations are a real or potential threat for psychological or physiological integrity of an individual. The underlying neurobiological substrates involved in these processes were substancially investigated through the use of animal models of stress. In this context, a crescent number of studies have used more naturalistic animal tests, such as the social defeat test. Regarding neurobiological substrates, it is known that the amygdala plays an important role in the modulation of defensive responses. This forebrain structure has several neurotransmitters and receptors with important implications in emotional states. In this context, the neuropeptide Corticotropin Releasing Factor (CRF) and its receptors, CRF1 and CRF2, have been recently investigated as an important modulatory system of defensive reactions to aversive situations. Activation CRF mechanisms in the amygdala has been postulated as a possible neurochemical substrate underlying the emotional disorders, especially anxiety disorders, induced by stress in humans. To study anxiety-related responses induced by stressors in animals, the elevated plus maze (EPM) test has been widely used. While previous studies have emphasized the role of CRF1 receptors in modulation of anxiety in rodents exposed to the EPM, the involvement of CRF2 receptors remains unclear. Few studies, however, have investigated the effects of CRF and CRF1 and CRF2 antagonists injected directly into the amygdala on the defensive responses in mice. In addition, several studies are needed to clarify the complex relationship between CRF neurotransmission of the amygdala in the etiology of anxiety disorders related to previous exposure to stress. This study investigated the role of CRF in the amygdala upon the defense reactions evaluated in the EPM in mice previously exposed to acute social defeat. Therefore, we carried out experiments to (i) characterize the effects of acute social defeat on behavior in the EPM and on the levels of plasma corticosterone; (ii) to investigate the effects of intraamygdala microinjection of CRF, CRF1 and CRF2 antagonists on the behavior of mice in the EPM and (iii) to investigate the effects of intra-amygdala microinjections of CRF1 and CRF2 antagonists on anxiety-related behaviors of mice pre-exposed to acute social defeat. Results showed that the exposure to acute social defeat stress produces anxiogenesis at short and long terms (i.e, assessed 5 min and 10 days after stress exposure), however short-term anxiety response is variable. Stress-short term effects are accompanied by increased plasma corticosterone levels. In addition, while intra-amygdala CRF increases anxiety, local injection of CRF1 (but not CRF2) receptor antagonists produced anxiolytic-like effects, suggesting a tonic role of CRF1 in the modulation of anxiety in mice exposed to the EPM. However, it was not possible to determine what is the role of CRF neurotransmission in the responses displayed by mice pre-exposed to social defeat and submitted to EPM. / O estresse é uma ameaça real ou potencial para a integridade psicológica ou fisiológica de um indivíduo e que resulta em respostas fisiológicas e/ou comportamentais. Os conhecimentos sobre estas respostas bem como sobre os substratos neurobiológicos envolvidos nestes processos só foram possíveis com o desenvolvimento de modelos animais de estresse. Dentre os vários modelos utilizados, destaca-se o modelo de derrota social por suas características mais etológicas. Em relação aos substratos neurobiológicos, é sabido que a amídala tem um importante papel na modulação de respostas defensivas. Esta estrutura encefálica possui diversos neurotransmissores e respectivos receptores com importantes implicações em estados emocionais, dentre eles o Fator de Liberação de Corticotropina (CRF). Os mecanismos de ação do CRF se dão por sua interação com os receptores CRF1 e CRF2. A ativação destes receptores na amídala tem sido postulada como um dos possíveis substratos neuroquímicos das alterações queocorrem nos transtornos comportamentais induzidos por estresse em humanos, destacando-se os transtornos de ansiedade por serem os mais prevalentes na população. Como ferramenta de estudo desses transtornos, temos o labirinto em cruz elevado (LCE), um dos mais populares modelos animais de ansiedade. Estudos no LCE apontam que o CRF1 modula a ansiedade, enquanto o papel do CRF2 não está claro. Poucos estudos, no entanto, têm investigado os efeitos do CRF, bem como de antagonistas para CRF1 e CRF2 injetados diretamente na amídala sobre as respostas defensivas de camundongos. Além disso, ainda se fazem necessários diversos estudos para entender a complexa relação entre a neurotransmissão CRFérgica da amídala na etiologia de transtornos de ansiedade relacionados a prévia exposição ao estresse. Sendo assim, o objetivo deste estudo é investigar o papel do CRF na amídala, nas reações de defesa avaliadas no LCE em camundongos previamente expostos ao estresse de derrota social agudo. Para tanto, realizou-se experimentos para (i) caracterizar os efeitos do estresse de derrota social agudo sobre o comportamento de camundongos no LCE e sobre os níveis de corticosterona plasmática; (ii) investigar os efeitos de microinjeções intra-amídala de CRF e de antagonistas CRF1 e CRF2 sobre os comportamentos de camundongos no LCE e (iii) investigar os efeitos de microinjeções intra-amídala de antagonistas CRF1 e CRF2 em camundongos pré-expostos ao estresse de derrota social agudo e submetidos ao LCE. Os resultados obtidos demonstram que o estresse de derrota social agudo é ansiogênico a curto e longo prazos, entretanto a resposta de ansiedade a curto prazo é variável. Estes mesmos efeitos a curto prazo são acompanhados por aumento do nível de corticosterona plasmática. Ainda demonstram que o CRF na amídala é ansiogênico e que há uma modulação tônica via CRF1, já o papel do CRF2 continua indeterminado. Entretanto, não foi possível determinar o papel da neurotransmissão CRFérgica nas respostas exibidas por camundongos pré-expostos ao estresse de derrota social e submetidos ao LCE.

abstract: Intermittent social defeat stress induces psychostimulant cross-sensitization, as well as long-lasting social avoidance behavior. Previous data reveal heightened expression of AMPA receptor (AMPAR) GluA1 subunits in rat ventral tegmental area (VTA), which occurs concurrently with social stress-induced amphetamine (AMPH) cross-sensitization. These studies described herein examined whether VTA GluA1 AMPARs are important for the behavioral consequences of social stress and investigated the role of the infralimbic (IL) to VTA pathway in the induction of these responses. Functional inactivation of GluA1 in VTA DA neurons prevented stress-induced AMPH sensitization without affecting social avoidance behavior, while GluA1 overexpression in VTA DA neurons mimicked the effects of stress on AMPH sensitization. Female rats were more sensitive to the effects of stress on AMPH administration than males, specifically during proestrus/estrus, which is characterized by higher circulating estradiol. Fluorescent immunohistochemistry revealed that females expressed higher GluA1 in VTA DA neurons as a result of intermittent social defeat stress, independent of estrus stage; by contrast, females during proestrus/estrus displayed higher tyrosine kinase receptor type 2 (TrkB) expression, which is the receptor for brain derived neurotrophic factor (BDNF), in VTA DA neurons, independent of stress exposure. Functional inactivation of GluA1 in VTA DA neurons prevented stress-induced AMPH sensitization and overexpression mimicked the effects of stress on AMPH sensitization. This suggests that BDNF-TrkB signaling may work concomitantly with GluA1 signaling in the VTA to drive sex-dependent differences in stress-induced locomotor sensitization effects. Optogenetic inhibition of the IL-VTA pathway in male rats prevented stress-induced AMPH sensitization compared to control animals. In addition, fluorescent immunohistochemistry displayed less Fos labeling in the nucleus accumbens (NAc) of rats with IL-VTA light inhibition compared to control animals. This suggests that the IL-VTA pathway plays a critical role in the induction of stress-induced sensitivity to AMPH, and blocking this pathway prevents mesolimbic DA signaling to the NAc. We conclude that IL glutamate projections onto GluA1-homomeric AMPA receptors in VTA DA neurons play a critical role in driving the stress-induced sensitization response in males and females. Therefore, GluA1 VTA DA neurons could potentially be a therapeutic target to prevent stress-induced drug susceptibility in the future. / Dissertation/Thesis / Doctoral Dissertation Neuroscience 2020

<p>Osteoarthritis (OA) is a leading cause of disability globally, with higher incidence in older people and lower socioeconomic status populations. The challenges health care systems face with management of the disease highlights the importance of OA research. Many studies examine possible risk factors of knee and hip OA including obesity, smoking, and alcohol consumption. Findings support that while obesity increases risk of knee OA, smoking is not a major risk factor. These extrinsic factors are, however, associated with lower socioeconomic status, and also with anxiety and depression disorders. Up to 30% of patients with chronic knee OA have described psychological stress and decreased quality of life due to debilitating pain, but the effects of psychological stress on development of knee OA has not been described.</p><p><br></p><p>At the cellular level, mechanosensitive cation channels in cartilage and bone, are involved with OA, but studies looking specifically at synovium and joint capsule are limited. Transient receptor potential (TRP) channels are upregulated in joint capsule in end-stage primary shoulder OA. We were unable to identify any previous studies evaluating Piezo channel expression in musculoskeletal soft tissues, but Piezo channel antagonism reduces chondrocyte death after mechanical injury. These findings suggest channels may help regulate joint responses to repetitive loading during training or work while also contributing to protective mechanisms within the musculoskeletal system. The overall objective of this research was to investigate factors that impact OA development or the disease phenotype. Two studies evaluated the following aims: 1) demonstrate the influence of chronic psychological stress on knee OA and overall systemic health, and 2) characterize the role of mechanosensitive channels in the joint capsule in OA. The first study used a mouse chronic social defeat model paired with destabilization of the medial meniscus (DMM) surgery to create a social stress scenario during OA development. We hypothesized chronic social defeat would exacerbate knee OA structural changes and systemic inflammation. The second study aimed to explore the role of mechanosensitive channels in joint capsule during OA development in the equine. Immunohistochemistry was performed on forelimb fetlock joint capsule from horses with varying degrees of lameness to first identify TRP and Piezo channel expression. Next, fibroblasts were isolated from the tissue to determine channel activity. We hypothesized that TRP and Piezo channels are required for normal homeostasis, but are dysregulated in OA and dysregulation contributes to fibrosis of the joint capsule. Joint capsule fibrosis leads to joint stiffening and reduced range of motion, two of the cardinal signs of OA.</p><p><br></p><p>The results of the first study showed OA was induced to a similar extent in both groups of mice that underwent DMM surgery. While anxiety- and depressive-like behaviors were exhibited by mice that underwent chronic social defeat episodes, unexpectedly, the majority of systemic inflammatory markers were not worse in mice with DMM and chronic social defeat compared to DMM alone. We were also able to show TRP and Piezo channel expression in one normal dorsal and palmar fetlock joint capsule sample, however, COVID-19 prevented further investigation. With our results we were able to conclude that while chronic social stress influences development of OA, in the current experiments, neither systemic inflammation nor structural signs of knee OA were worse with chronic social stress. We hope that exploration of OA through these two studies will help us understand how the disease contributes to overall systemic dysfunction while also providing a baseline for future development of TRP and Piezo channel modulators to prevent joint pathologies.</p>

Chronic stress and dysfunction of the serotonergic system in the brain have been considered two of the major risks for development of depression. In this study, adult Fischer 344 rats were subjected to a regimen of chronic social defeat (CSD). To mimic stressful conditions, some rats were not exposed to CSD, but instead treated with corticosterone (CORT) in oral solution while maintained in their home cage. Protein levels of the serotonin transporter (SERT) in the dorsal raphe nucleus (DRN), hippocampus, frontal cortex, and amygdala were examined by Western blotting or immunofluorescence staining. The results showed that CSD up-regulated SERT protein levels in the DRN, hippocampus, frontal cortex, and amygdala regions. This up-regulation was abolished or prevented by adrenalectomy, or treatment with antagonists of corticosteroid receptors mifepristone and spironolactone, alone or in combination. Similarly, up-regulated SERT protein levels in these brain regions were also observed in rats treated with oral CORT ingestion, which was analogously prevented by treatment with mifepristone and spironolactone. Furthermore, both CSD- and CORT-induced up-regulation of SERT protein levels in the DRN and three brain regions were attenuated by simultaneous treatment with fluoxetine, an antidepressant that specifically inhibits serotonin reuptake. The results indicate that up-regulation in SERT protein levels in the DRN and forebrain limbic structures caused by CSD regimen was mainly motivated by CORT through corticosteroid receptors. The present findings demonstrate that chronic stress is closely correlated with the serotonergic system by acting on the regulation of the SERT expression in the DRN and its projection regions, which may contribute to the development of depression. Chronic stress and dysfunction of the serotonergic system are etiologically related to depression. In an attempt to explore their interaction, we found that chronic social defeat upregulated expression of serotonin transporter in the DRN and the projection regions, which may induce an alteration of serotonin transformation in the brain. This interaction may account for the development of this disease.