O estresse parece favorecer a hiperalgesia e alodinia, podendo estar associados à alteração da função muscular mastigatória. Alterações morfofisiológicas em músculos da mastigação induzidos pela alteração oclusal associado ao estresse crônico ainda são escassas na literatura. Este estudo investigou os efeitos do estresse crônico repetido em músculo masseter superficial e profundo de ratos submetidos ou não à exodontia unilateral no ganho do peso dos animais, nas alterações morfológicas (HE, MET), histoquímicas (NADH, SDH e ROS), imunoistoquímicas (laminina e CD31), atividade de MMP-2, -9 e infiltração de neutrófilos (MPO). Vinte ratos (machos, 200g) foram alocados em quatro grupos (n=5): controle (GC), exodontia unilateral (GM), estresse crônico repetido (GE), exodontia associado ao estresse crônico repetido (GME). GE e GME foram submetidos a 10 dias de protocolo de estresse crônico repetido (2 horas diárias) a partir do 14º dia após a exodontia. Houve uma diminuição significativa no ganho de peso dos animais GE e GME. Não foram observadas alterações nos níveis de MMPs e na infiltração de neutrófilos no feixe superficial dos diferentes grupos. GE, GM e GME demostraram alterações morfofisiológicas, ultraestruturais e histoquímicas no feixe profundo, com características específicas e distintas de GC; GE apresentou as maiores alterações. Conclui-se que a exodontia e sua associação ao estresse foram responsáveis por discretas alterações morfofisiológicas no músculo masseter de ratos, contudo o estresse crônico repetido causou modificações morfofisiológicas e ultraestruturais significantes, sendo responsável também pela alteração no peso dos animais. / Stress seems to favor the hyperalgesia and allodynia, which may be related with altered masticatory muscle function. Morphological and physiological changes in the masticatory muscles induced by occlusal alteration associated with chronic stress are still scarce in the literature. This study investigated the effects of repeated chronic stress in superficial and deep masseter muscle of rats with or without the extraction unilateral weight gain of animals and morphological changes (HE MET), histochemical (NADH, SDH and ROS), immunohistochemical (laminin and CD31), MMP-2, -9 activities and neutrophil infiltration (MPO). Twenty rats (male, 200g) were allocated into four groups (n=5): control group (CG), unilateral exodontia (MG), repeated chronic stress (EG), extodontia and repeated chronic stress (MEG). EG and MEG were submitted to 10 days of repeated chronic stress protocol, 2 hours daily, from the 14th day after the extraction. There was a significant decrease in weight gain of animals EG and MEG. No changes were observed in the levels of MMPs and neutrophil infiltration among different groups. EG, MG and MEG have shown morphophisyological, ultrastructural and histochemical changes with specific characteristics and distinct GC GE presenting the higher changes. We conclude that the exodontia and its association to stress were responsible for discrete morphophysiological changes in the masseter muscle of rats, however repeated chronic stress caused significant morphophysiological and ultrastructural changes, being also responsible for change in weight of the animals.

abstract: The present series of studies examined whether a novel implementation of an intermittent restraint (IR) chronic stress paradigm could be used to investigate hippocampal-dependent spatial ability in both sexes. In experiments 1 and 2, Sprague- Dawley male rats were used to identify the optimal IR parameters to assess spatial ability. For IR, rats were restrained for 2 or 6hrs/day (IR2, IR6, respectively) for five days and then given two days off, a process that was repeated for three weeks and compared to rats restrained for 6hrs/d for each day (DR6) and non-stressed controls (CON). Spatial memory was tested on the radial arm water maze (RAWM), object placement (OP), novel object recognition (NOR) and Y-maze. The results for the first two experiments revealed that IR6, but not IR2, was effective in impairing spatial memory in male rats and that task order impacted performance. In experiment 3, an extended IR paradigm for six weeks was implemented before spatial memory testing commenced in male and female rats (IR- M, IR-F). Unexpectedly, an extended IR paradigm failed to impair spatial memory in either males or females, suggesting that when extended, the IR paradigm may have become predictable. In experiment 4, an unpredictable IR (UIR) paradigm was implemented, in which restraint duration (30 or 60-min) combined with orbital shaking, time of day, and the days off from UIR were varied. UIR impaired spatial memory in males, but not females. Together with other reports, these findings support the interpretation that chronic stress negatively impairs hippocampal-dependent function in males, but not females, and that females appear to be resilient to spatial memory deficits in the face of chronic stress. / Dissertation/Thesis / Masters Thesis Psychology 2019

The unfolded protein response (UPR) is activated by protein misfolding stress in the endoplasmic reticulum (ER). The UPR is a transcriptional program that aims to maintain ER folding capacity, where imbalances between protein load and processing ability is termed ER stress. Signal transduction of the UPR begins with 3 ER-resident transmembrane sensors: PERK, IRE1 and ATF6. All sensors initiate downstream signaling cascades which culminate in improved protein folding, transcriptional upregulation of genes encoding ER chaperones, and mechanisms to reduce translational and transcriptional ER load, therefore re-establishing ER homeostasis. The signaling cascades of each sensor are distinct but cooperative, and involve a significant amount of crosstalk, feedback and overlap. Indeed, there are many pathological and physiological conditions have an effect on ER protein burden, and therefore on activation of the UPR. Increases in protein load in professional secretory cells, hypoxic conditions in a tumor mass, obesity all induce cause changes in the ER folding environment. Although we understand how the UPR contributes to relieve ER stress under acute conditions (e.g. pharmacological treatment) much less is understood about the contributions to physiological processes and chronic stress conditions. Our overall goal was to understand how the UPR is activated during physiological settings, the mechanisms it uses to maintain folding capacity under these setting and the specific components responsible for adapting the response to various stresses. We first decided to understand a chronic stress from a transgenic approach. By creating a knockout mouse, the genetic deletion functions as a stress and we can understand its physiological role. By compounding two genetic deletions in UPR components (ATF6α and p58IPK) we provide evidence for the developmental role these components play. Homozygous deletion ATF6α bears no gross histological phenotype yet causes synthetic lethality when combined with p58IPK deletion. This also reveals that the UPR is able to adapt to genetic impairment of protein folding in vivo. Next, to better understand these chronic states, we established an experimentally tractable chronic stress treatment in vivo. Our treatment suppressed ATF6α dependent chaperone expression through an mRNA degradative mechanism, which led to long term changes in UPR expression. We determined that chronic conditions can change the sensitivity of the UPR to ER stress, potentially as an adaptive consequence. We also showed that sensitivity to ER stress can be changed during chronic stress. Finally we simulated the UPR in a computational ordinary differential equation (ODE) model in order to determine how various stresses and component interactions determine the output of the UPR. We built a series of equations to describe the UPR signaling network, entrained it on experimental data and refined it through the use of transgenic knockout cells. Our model was robust enough to recreate experimental measurements of UPR components when tested in parallel with knockout cells. We found that stress sensitivity is dependent on the crosstalk and negative feedback connections of the UPR. This study has enhanced our understanding of activation of the UPR under non-acute settings. It demonstrates that the UPR is a signaling hub with a broad output range that is capable of handling a variable degree of insults because of the intrinsic properties of the signaling network. This provides a better understanding for the contributions of the UPR to physiological stresses and certain chronic diseases.

During the critical period of CNS development, dendritic architecture is shaped, in part, by activity-dependent stabilization and elimination of branches. This restructuring is partly dependent on the subunit composition of glutamate receptors in a manner that is both regionally specific and temporally regulated. We used primary cultures of rodent hippocampal neurons to investigate the consequences for hippocampal dendrite development when the glutamate ?-amino-3-hydroxy-5-methyl-4-isoxazoleproprionic acid receptor (AMPAR) subunit composition was altered. Overexpression of the AMPAR subunits GluR1 or GluR2 differentially modified hippocampal dendrite architecture. We investigated signaling pathways known to be involved in activity-dependent circuit development as possible downstream effectors of AMPA-mediated morphogenesis. We identified extracellular signal regulated kinase (ERK) 1/2 as a potential candidate of GluR1-mediated dendrite outgrowth. We found that levels of docosahexaenoic acid (DHA) and a DHA-derived bioactive metabolite, neuroprotectin D1 (NPD1) are differentially regulated by GluR1 and GluR2. DHA, but not NPD1, induced extensive dendritic branching and outgrowth. Overexpression of 15 lipoxygenase 1 (15LOX1), the enzyme responsible for conversion of DHA to NPD1, interrupted outgrowth mediated by GluR1 overexpression. In order to investigate molecular mechanisms that regulated neural circuitry outside of the critical period of CNS development, we examined dendrite morphology across the CNS in response to chronic variable stress (CVS). We found wide-spread changes in circuits implicated in neurocognitive dysfunctions associated with chronic stress, and observed substantial dendritic plasticity in the adult brain. / acase@tulane.edu

According to recent nationwide surveys there have been dramatic increases in stress and serious stress-related health problems among Swedish adolescents. The aims of the present thesis were to investigate the prevalence of perceived chronic stress among sixteen-year-old adolescents who attended their first year at high-ranking high schools in the Stockholm area, to examine if factors that have been found to be relevant in the development of chronic stress among adults also contribute to chronic stress among adolescents, and to investigate the associations between chronic stress and sleep-related variables and academic achievement. Data were collected by means of self-report instruments. The main outcomes showed that a substantial number of individuals (30%) reported to perceive severe stress symptoms. The highest degree was perceived by 8% who suffered from symptoms to an extent which otherwise only has been observed among adult clinical burnout patients. Besides, 15% reported severe stress symptoms throughout high school and were thus considered to suffer from chronic stress. Perceived high demands, low global self-esteem, sleep disturbances and low social support were crucial factors in explaining stress symptoms. Another finding showed that perceived chronic stress was associated with adolescents’ academic achievement. A closer look at the potential contribution of sleep-related variables to academic achievement revealed further that, apart from severe stress symptoms, social jetlag was involved in predicting variation in academic achievement. Overall, the findings indicated that there is a high prevalence of chronic stress among adolescents. The results should be taken seriously, as young people’s sense of well-being during this period impacts both their successful transition into adulthood and their life course. Future research options and potential practical applications in terms of chronic stress prevention were suggested and discussed. / <p>At the time of the doctoral defense, the following paper was unpublished and had a status as follows: Paper 3: Submitted.</p>

The physiological responses to stressor exposure can be broadly grouped into the organismal and the cellular stress responses. The organismal stress response involves the release of hormones into general circulation, while the cellular stress response involves the synthesis of proteins, the most important being the heat shock proteins (HSPs), which play a role in maintaining protein homeostasis. Elevated HSP70 expression in response to stressors has been demonstrated in trout (Oncorhynchus mykiss) red blood cells (RBCs). The ease of repeated sampling of blood suggests the possibility of using this tissue as a non-lethal marker of cellular stress in fish. This study tested the hypothesis that stressor exposure will elevate HSP70 expression in trout RBCs and the role of stress hormones in mediating this response. Acute heat shock exposure (+12oC) significantly elevated plasma cortisol, glucose and lactate levels in heat shocked fish over 24 h. A tissue-specific response was seen in HSP70 expression in liver, brain, gill and RBCs. To enable measurement of RBC HSP70 concentrations, an enzyme-linked immunosorbent assay (ELISA) was developed using a commercially available rabbit anti-salmon HSP70 and a recombinant chinook salmon (Oncorhynchus tshawytscha) HSP70. To determine effects of chronic exposure, two studies were conducted exposing trout to either cadmium (0, 0.75 or 2.0 µg/L over 28 d) or municipal wastewater effluent (0, 20 or 90% over 14 d). However, neither exposure elicited a significant HSP70 response. Effects of stress hormones on RBC HSP70 levels were tested by exposing cells in vitro to either cortisol (10 and 100 ng/mL) or epinephrine (10 nM) with or without heat shock. Heat shock elevated HSP70 content in trout RBCs but no modulation by stress hormones was seen. It was shown for the first time that RBCs release HSP70 content into the medium in response to an acute heat shock and this release is attenuated by stress hormones. Overall, HSP70 levels in RBCs have the potential to be a reliable non-lethal marker of acute cellular stress effects in fish. The release of HSP70 from RBCs leads to the hypothesis that HSP70 may also have an extracellular role in fish, and warrants further study.

In certain species, puberty is thought to be a period of susceptibility to various stressors, resulting in pathological behavioral and physiological changes subsequent to exposure during this period. However, juvenile male golden hamsters appear to be fairly resilient to pubertal stress, as compared to adult hamsters and many other species. In these experiments, repeatedly stressed juvenile male hamsters were found to be avoidant of aggressive adult male social stimuli, but did not display anxious behavior outside of a social context. In addition, several long-term changes in neural activity were associated with social stress during early puberty. The medial preoptic area and medial preoptic nucleus, and ventral tegmental area showed decreased neural activity in subjugated juveniles than in naïve individuals. Since these brain areas are involved in the expression of motivated behaviors, specifically sexual behavior, and reward pathways, we next investigated sexual behavior in virgin juveniles. When placed in a confined space with receptive females, consummatory behavior in subjugated juveniles was similar to those observed in naive juveniles. Appetitive aspects of sexual behavior were also tested in a Y-maze to allow subjects to choose whether to approach a social stimulus. When given a choice between a sexually receptive and non-receptive female social stimulus, socially stressed individuals showed anxiety related behaviors and did not show a preference. However, naïve hamsters preferred the non-receptive female. Interestingly, this effect was less significant in naïve animals tested during late puberty and early adulthood, and a preference for sexually receptive females was not observed. In addition, stressed hamsters tested with harnessed females at mid-puberty were slower to approach females, indicating altered motivation to approach adult conspecifics. This research is unique in that it is the first to suggest the disconnect between the development of consummatory and appetitive aspects of sexual behavior. Together, these data examine the effects of stress on the development of pubertal social behaviors. / text

The physiological responses to stressor exposure can be broadly grouped into the organismal and the cellular stress responses. The organismal stress response involves the release of hormones into general circulation, while the cellular stress response involves the synthesis of proteins, the most important being the heat shock proteins (HSPs), which play a role in maintaining protein homeostasis. Elevated HSP70 expression in response to stressors has been demonstrated in trout (Oncorhynchus mykiss) red blood cells (RBCs). The ease of repeated sampling of blood suggests the possibility of using this tissue as a non-lethal marker of cellular stress in fish. This study tested the hypothesis that stressor exposure will elevate HSP70 expression in trout RBCs and the role of stress hormones in mediating this response. Acute heat shock exposure (+12oC) significantly elevated plasma cortisol, glucose and lactate levels in heat shocked fish over 24 h. A tissue-specific response was seen in HSP70 expression in liver, brain, gill and RBCs. To enable measurement of RBC HSP70 concentrations, an enzyme-linked immunosorbent assay (ELISA) was developed using a commercially available rabbit anti-salmon HSP70 and a recombinant chinook salmon (Oncorhynchus tshawytscha) HSP70. To determine effects of chronic exposure, two studies were conducted exposing trout to either cadmium (0, 0.75 or 2.0 µg/L over 28 d) or municipal wastewater effluent (0, 20 or 90% over 14 d). However, neither exposure elicited a significant HSP70 response. Effects of stress hormones on RBC HSP70 levels were tested by exposing cells in vitro to either cortisol (10 and 100 ng/mL) or epinephrine (10 nM) with or without heat shock. Heat shock elevated HSP70 content in trout RBCs but no modulation by stress hormones was seen. It was shown for the first time that RBCs release HSP70 content into the medium in response to an acute heat shock and this release is attenuated by stress hormones. Overall, HSP70 levels in RBCs have the potential to be a reliable non-lethal marker of acute cellular stress effects in fish. The release of HSP70 from RBCs leads to the hypothesis that HSP70 may also have an extracellular role in fish, and warrants further study.

Over 50% of the world’s human population resides in urban centres, and this is expected to increase as the global human population grows and people migrate from non-urban to urban centres. Concentrated in these urban areas are anthropogenic disturbances that impose additional challenges on wildlife compared to their non-urban counterparts. These challenges can be stress provoking. Through the release of corticosterone (CORT) reptiles can adapt to these stressors, physiologically and behaviourally, both in the short- and long-term. To investigate the relationships between stress activation and defensive tactics in wild urban Northwestern Gartersnakes (Thamnophis ordinoides) I conducted visual encounter surveys, along edge-focused transects, following a semi-constrained random sampling method. I sampled snakes at five sites, each with a different level of anthropogenic disturbance, in the Greater Victoria Area, BC. I sampled blood, observed anti-predator behaviour, and collected data on characteristics of snakes. The most disturbed site (with the most people, pets, and natural predators) also had the most snakes: those snakes also had highest H:L values (a proxy of CORT) in their blood compared to the other populations. Nevertheless, none of the snakes had H:L values that indicated chronic stress. Stress physiology was not correlated with anti-predator behaviour. More important to anti-predator behaviour was the size, sex/reproductive condition, and cloacal temperature of snakes. Although anthropogenic development can reduce habitat quality for some reptiles, Northwestern Gartersnakes coexist with recreationists at many sites in the District of Saanich. A multi-disciplinary approach is of paramount importance to understand the full effect of anthropogenic influences on wildlife. / Graduate / 0433 / 0329

Resumo: O estresse é entendido como um processo complexo e multidimensional, cuja resposta adaptativa é vista como um processo dinâmico no qual mecanismos fisiológicos do indivíduo mudam continuamente para ajustarem-se ao ambiente. Esse fator pode contribuir para alterações na função cardíaca, associada, ou não, ao trânsito de Ca2+ e a alteração vascular devido ao aumento da atividade da via l-arginina/óxido nítrico. A obesidade é uma doença complexa, caracterizada pelo acúmulo excessivo de tecido adiposo, levando a disfunções cardíacas e vasculares, que podem estar envolvidas com alteração no fluxo de Ca2+ e comprometimento da resposta vasodilatadora. O estresse é considerado um fator ambiental, portanto, responsável por alterações no balanço energético e peso corpóreo, uma vez que o peso corpóreo é a interação entre fatores genéticos e ambientais. Devido à escassez de estudos que avaliam o estresse crônico e dieta hipercalórica nas alterações cardiovasculares, e considerando a hipótese de que esta associação é capaz de atenuar o desenvolvimento da obesidade e intensificar as alterações cardiovasculares, o objetivo do presente trabalho foi confirmar essa hipótese em animais submetidos à dieta hipercalórica e ao estresse crônico. Ratos machos Wistar, divididos em quatro grupos: DN: dieta normocalórica; DN/Es: dieta normocalórica e submetidos ao estresse crônico; DH: dieta hipercalórica; DH/Es: dieta hipercalórica e submetidos ao estresse crônico, foram avaliados quanto aos perfis nutricionais, metabólicos e a remodelação cardiovascular. Os dados demonstraram que o estresse crônico impediu o desenvolvimento da obesidade, e em oposição à hipótese, o estresse crônico melhorou a função cardíaca e vascular, independente do tipo de dieta utilizada / Abstract: The stress is a complex and multidimensional process, the adaptive response is a dynamic process, in which individual physiologic mechanisms change to adjust to the environment, that causes alteration in the cardiac function, involved, or not, with Ca2+ flux and vascular alterations due to the increase of the l-arginine/nitric oxide pathway activity. The obesity is a complex disease, characterized by the excess of adipose tissue that causes cardiac and vascular dysfunction, involved with Ca2+ flux alterations and injury the vasodilating response. The stress is considered an environmental factor, therefore, it is responsible for the energetic balance alterations and weight gain, once the body weight is an interaction between genetic and environmental factors. There are few studies about chronic stress associated with the hypercaloric diet that evaluate the cardiovascular alterations, and considering the idea that this association can decrease the obesity development and increase the cardiovascular alterations, the aim of this research was evaluate the cardiovascular alterations and the obesity development in animals submitted to the hypercaloric diet and chronic stress. Male Wistar rats were separated into four groups: DN: standard diet; DN/Es: standard diet and chronic stress; DH: hypercaloric diet; DH/Es: hypercaloric diet and chronic stress were evaluated in relation to the nutritional and metabolic profile and cardiovascular remodellating. The data show that the chronic stress inhibited the obesity development, and different of the initial idea, the chronic stress improved the cardiac and vascular function, in both diets / Orientador: Sandra Cordellini / Coorientador: Antônio Carlos Cicogna / Banca: Rita de Cássia Aleixo Tostes Passaglia / Banca: André Sampaio Pupo / Mestre