Chronic stress and elevated glucocorticoids (GCs), the major stress hormones, are risk factors for Alzheimer’s disease (AD) and promote AD pathomechanisms in animal models. These include overproduction of synaptotoxic amyloid-β (Aβ) peptides and intraneuronal accumulation of microtubule-associated protein Tau. Tau accumulation is linked to downregulation of the small GTPase Rab35, which mediates Tau degradation via the endolysosomal pathway. Whether Rab35 is also involved in stress/GC-induced Aβ overproduction remains an open question. Here, I find that hippocampal Rab35 levels are decreased not only by stress/GCs, but also by aging, another AD risk factor. Moreover, I show that Rab35 negatively regulates Aβ production by sorting amyloid precursor protein (APP) and β-secretase (BACE1) out of the endosomal network, where they interact to produce Aβ. Interestingly, Rab35 coordinates distinct intracellular trafficking events for BACE1 and APP, mediated by its effectors OCRL and ACAP2, respectively. Additionally, I show that Rab35 overexpression prevents the amyloidogenic trafficking of APP and BACE1 induced by GCs. Finally, I begin to investigate how GCs and/or Rab35 affect the intercellular spread of Aβ and Tau through exosomes. I describe methods for purifying exosomes and measuring their secretion from neurons, astrocytes, and microglial cells in order to determine the effects of stress/GCs and Rab35 on this process. These studies identify Rab35 as a key regulator of Alzheimer’s disease-related protein trafficking, and suggest that its downregulation contributes to stress- and AD-related pathomechanisms.

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Indiana University-Purdue University Indianapolis (IUPUI) / Phosphorylation of eukaryotic initiation factor 2 (eIF2) is an important mechanism regulating global and gene-specific translation during different environmental stresses. Repressed global translation by eIF2 phosphorylation allows for cells to conserve resources and elicit a program of gene expression to alleviate stress-induced injury. Central to this gene expression program is eIF2 phosphorylation induction of preferential translation of ATF4. ATF4 is a transcriptional activator of genes involved in stress remediation, a pathway referred to as the Integrated Stress Response (ISR). We investigated whether there are additional transcription factors whose translational expression is regulated by eIF2 kinases. We found that the expression of the transcriptional regulator ATF5 is enhanced in response to many different stresses, including endoplasmic reticulum stress, arsenite exposure, and proteasome inhibition, by a mechanism requiring eIF2 phosphorylation. ATF5 is regulated by translational control as illustrated by the preferential association of ATF5 mRNA with large polyribosomes in response to stress. ATF5 translational control involves two upstream open reading frames (uORFs) located in the 5′-leader of the ATF5 mRNA, a feature shared with ATF4. Mutational analyses of the 5′-leader of ATF5 mRNA fused to a luciferase reporter suggests that the 5′-proximal uORF1 is positive-acting, allowing scanning ribosomes to reinitiate translation of a downstream ORF. During non-stressed conditions, when eIF2 phosphorylation is low, ribosomes reinitiate translation at the next ORF, the inhibitory uORF2. Phosphorylation of eIF2 during stress delays translation reinitiation, allowing scanning ribosomes to bypass uORF2, and instead translate the ATF5 coding region. In addition to translational control, ATF5 mRNA and protein levels are significantly reduced in mouse embryo fibroblasts deleted for ATF4, or its target gene, the transcriptional factor CHOP. This suggests that ISR transcriptional mechanisms also contribute to ATF5 expression. To address the function of ATF5 in the ISR, we employed a shRNA knock-down strategy and our analysis suggests that ATF5 promotes apoptosis under stress conditions via caspase-dependent mechanisms. Given the well-characterized role of CHOP in the promotion of apoptosis, this study suggests that there is an ATF4-CHOP-ATF5 signaling axis in the ISR that can determine cell survival during different environmental stresses.

A recent evolutionary theory hypothesizes that there are two primary biobehavioral profiles of stress responding. Labeled "hawk" and "dove," each is characterized by divergent patterns of autonomic nervous system and neuroendocrine system activations in response to stress as well as distinct affective and behavioral tendencies. These profiles are prominent in a number of species, and it has been hypothesized that hawk-like and dovelike responses to stress may, in part, explain variability in stress-related health outcomes. This study is a preliminary investigation of hawk and dove biobehavioral profiles in humans. Participants included 73 Midwestern university students recruited from undergraduate-level psychology classes. Upon completion of a stressor task, participants answered questions regarding their psychological experiences during and immediately following the task and reported their emotions and health-related behaviors over the past several weeks. Physiological measures of cortisol and high frequency heart rate variability reactivity were used to identify relatively hawk-like and dove-like responders. Associations between patterns of physiological responding and emotional and behavioral responses were tested. The results showed mixed support for the existence of hawk and dove biobehavioral profiles in humans.

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Aerobic exercise is frequently recommended as a stress management strategy. This study examined the influence of 30 minutes of aerobic exercise in reducing cardiovascular reactivity to a subsequent stressor compared to a nonexercise attention control condition in 16 males (<u>m</u> age = 21 yrs; moderate activity level; positive family history for hypertension) using a within subject design. The order of experimental conditions was counterbalanced across subjects. During the exercise condition, subjects pedaled for 30 minutes on a bicycle ergometer at 59% ± 7.2 of their maximum (<u>m</u> V0₂= 48.5 ml/kg/min ± 10.7). subjects completed a physical activity questionnaire and anthropometric measurements during the 30 minute attention control condition. Forty-five minutes after the exercise or attention control sessions, BP, HR, stroke volume (SV) and total peripheral resistance (TPR) were measured at baseline, during a pre-stressor anticipation period, and during the cold pressor test. stroke volume was measured with impedance cardiography and total peripheral resistance (TPR) was estimated. SBP was significantly lower during the second minute of the anticipation period (F [1,29] = 4.75, p = .04) forty-five minutes after exercising compared to the attention control condition. There were no significant differences between conditions for DBP, TPR or SV. These findings suggest that acute aerobic exercise may moderate SBP response to a subsequent non-exercise stressor. / Master of Science

Call number: LD2668 .T4 1985 D69 / Master of Science

Call number: LD2668 .T4 1985 D37 / Master of Science

Dissertation (PhD)--University of Stellenbosch, 2004. / ENGLISH ABSTRACT: Stress related to chronic exercise affects both the immune and endocrine systems, but there are still many issues that are poorly understood, particularly effects of stress on the functional capacity of immune cells. This thesis probed some of these issues using physiological models of physical and psychological stress. Both exercise training stress and chronic psychological stress in human subjects were shown to result in an up-regulation of spontaneous reactivity of white blood cells in vitro, using two different assays, namely a) a peripheral blood mononuclear cell (PBMC) culture assay measuring immune cell responsiveness and b) a relatively new flow cytometry technique for assessing activation status of cells by their expression of the surface marker CD69, in a lymphocyte subpopulation-specific manner. An up-regulation of immune cell activation in the absence of an additional stressor was associated with a decreased capacity to mount a response to a subsequent mitogen stimulus in vitro after chronic psychological stress and acute, extreme exercise stress. Another novel finding was that cortisol high-responders to chronic psychological stress exhibited a higher spontaneous reactivity of both CD4+ and CD8+ lymphocytes when compared to cortisol low-responders. This result indicates that chronic exposure to cortisol may decrease its usual inhibitory effect on spontaneous T lymphocyte responsiveness. After optimisation of an animal model of mild, psychological stress, we demonstrated (using an IL-6 antibody) that IL-6 is necessary for a full-blown cortisol response to chronic, intermittent mild stress. Results also suggest that IL-6 plays a role in regulation of its own secretion by PBMCs in response to a stressor, by maintaining the production of IL-1β in the face of stress. Basal serum corticosterone concentration was shown to be the main determinant of the magnitude of mitogen-stimulated PBMC secretion of IL-6 in vitro in the stress-free controls. However, after blocking of IL-6 in vivo, IL-1β was identified as a major regulator of IL-6 secretion by mitogen-stimulated PBMCs in vitro, independently of the presence or absence of stress. The implications of these novel findings are that proinflammatory cytokines are sensitively regulated during mild stress.Mean serum cortisol concentration at rest was not a useful tool to assess chronic exercise stress after training intervention. However, classification of athletes at baseline into two groups according to their resting serum cortisol concentration illustrated two distinct patterns for the responses of both cortisol and the cortisol:testosterone ratio to chronic stress. These studies on the effects of chronic stress on parameters of the endocrine stress-axis and the immune system led to the following main conclusions: a) chronic exposure to cortisol results in a decreased inhibition of spontaneous immune cell activity at rest, b) this increased spontaneous activation of immune cells at rest in the absence of a stressor, is associated with a suppression of immune capacity to respond to a subsequent challenge, c) the latter finding is not evident under stress-free conditions where cortisol promoted immune cell IL-6 secretion, and d) IL- 1β and IL-6 are involved in the regulation of each others’ secretion. / AFRIKAANSE OPSOMMING: Chroniese oefening-verwante stres beïnvloed beide the immuun- en endokriene sisteme, maar daar is nog baie aspekte wat swak begryp word, veral m.b.t. die effekte van stres op die funksionele kapasiteit van immuunselle. Hierdie tesis het sommige van dié vraagpunte ondersoek deur gebruik te maak van fisiologiese en psigologiese stres. Beide oefening program-verwante stres en chroniese psigologiese stres in proefpersone het ‘n op-regulering van spontane witbloedselreaktiwiteit in vitro tot gevolg gehad, wat d.m.v twee verskillende metodes aangetoon is, naamlik a) ‘n perifere bloed mononukluêre selkultuur (PBMS-kultuur) bepaling van immuunsel reaktiwiteit en b) ‘n relatief nuwe vloeisitometriese tegniek vir die assessering van aktiveringsstatus van selle, deur hul uitdrukking van die oppervlakmerker CD69, op ‘n limfosiet subpopulasie-spesifieke wyse. ‘n Opregulering van immuunselaktiwiteit in die afwesigheid van ‘n addisionele stressor is geassosieer met ‘n verlaagde kapsiteit om te reageer op ‘n latere mitogeniese prikkel in vitro, na chroniese psigologiese stres en akute, erge oefeningstres. Nog ‘n nuwe bevinding was dat kortisol hoog-respondeerders, in reaksie op chroniese psigologiese stres, ‘n hoër spontane reaktiwiteit van beide CD4+- and CD8+-limfosiete toon in vergelyking met kortisol laagresopndeerders. Hierdie bevinding toon aan dat chroniese blootstelling aan kortisol die inhiberende effek daarvan op spontane reaktiwiteit van T-limfosiete verminder. Na optimalisering van ‘n rotmodel van gematigde, psigologiese stres, het ons gedemonstreer (deur gebruik te maak van ‘n IL-6 teenliggaam) dat IL-6 nodig is vir ‘n volledige kortisolreaksie op chroniese, onderbroke, gematigde stres. Die resultate dui daarop dat IL-6 ‘n rol in die regulering van sy eie sekresie deur PBMSe in reaksie tot ‘n stressor speel, deur die handhawing van produksie van IL-1β in die teenwoordigheid van stres. Basale serum kortisolkonsentrasie is as die belangrikste beslissende faktor in die omvang van mitogeengestimuleerde PBMS sekresie van IL-6 in vitro in die stresvrye kontroles aangedui. Na blokkering van IL-6 in vivo, is IL-1β egter as ‘n belangrike reguleerder van IL-6 sekresie deur mitogeen-gestimuleerde PBMSe in vitro geïdentifiseer, onafhanklik van die teenwoordigheid of afwesigheid van stres. Die implikasie van hierdie nuwe bevindinge is dat proinflammatoriese sitokiene tydens gematigde stres sensitief gereguleer word.Die gemiddelde serum kortisolkonsentrasie in ‘n rustende toestand was nie ‘n gepaste instrument om chroniese oefeningstres na ‘n oefenprogram-ingreep te assesseer nie. Na basislyn klassifikasie van atlete in twee groepe volgens hul rustende serum kortisolkonsentrasie, is twee afsonderlike patrone vir die reaksie van beide kortisol en die kortisol:testosteroon verhouding egter aangetoon. Hierdie studies rakende die effekte van chroniese stres op parameters van die endokriene stres-as en die immuunsisteem het tot die volgende vernaamste gevolgtrekkings gelei: a) chroniese blootstelling aan kortisol het ‘n verlaagde inhibisie van spontane immuunselaktiwiteit tydens rustende toestande tot gevolg, b) hierdie verhoogde spontane aktivering van immuunselle tydens ‘n rustende toestand word geassosieer met ‘n onderdrukking van immuunkapasiteit om te reageer op ‘n daaropvolgende prikkel, c) laasgenoemde bevinding is nie sigbaar tydens stresvrye toestande, wanneer kortisol IL-6 sekresie bevorder, nie en d) IL- 1β en IL-6 is betrokke by die regulering van mekaar se sekresie.