Non-compensable cold exposure represents a potentially deadly threat to humans, as we lack highly specialized organs and mechanisms necessary to maintain our optimal core temperature of ~37°C. Repeated exposures to cold have been shown to induce protective physiological changes in cold responses through a process known as cold acclimatization (natural) or acclimation (in laboratory). The purpose of this thesis was to determine what physiological changes occur following an intense 7 day, 14°C cold water immersion acclimation protocol, during both non-compensable (Chapter 2) and compensable cold exposures (Chapter 3). This includes identifying changes in the contributions of the shivering (ST) and non-shivering (NST) thermogenic pathways to overall heat production. ST and NST changes were quantified via electromyography and indirect calorimetry, respectively. This 7 day cold water acclimation protocol resulted in a decrease in cooling rate, a significant increase in mean esophageal core temperature, a decrease in peak heart rate following immersion, and increased thermal comfort from day 1 to day 7 of the 1h 14°C cold water immersions. Further to these findings, changes in ST and NST were measured pre- and post-acclimation with a standardized compensable cold protocol using a liquid conditioned suit (LCS) which lowered Tskin to 26°C for 2.5h. The cold acclimation protocol resulted in a ~38% decrease in mean shivering over the 2.5h without any change in thermogenic rate from pre- to post-cold acclimation. In addition, no significant difference in fuel selection was observed. These results indicate that the short, intense cold acclimation protocol did result in a substantial change in the contribution of ST and NST to total heat production which could increase cold tolerance by reducing involuntary muscle contractions during ST.

Both white and brown adipose tissues exhibit extensive vascularity. Increased angiogenesis in brown adipose tissue (BAT) is crucial for brown fat activation and thermogenesis in animals during cold acclimation. BAT can be similarly activated by food intake to generate heat through cellular respiration, in a process known as diet induced thermogenesis. Vascular endothelial growth factor (VEGF) is a potent angiogenic factor that regulates both pathological and physiological angiogenesis and can stimulate cell proliferation, migration, survival and vessel permeability. However, VEGF has also been shown to affect an increasing number of non-vascular cells such as skeletal muscle and kidney podocytes. The expression and function of VEGF in white and brown adipocytes are not fully understood. We have previously shown that the expression of VEGF is concomitantly regulated with skeletal muscle differentiation. Here we show that VEGF is expressed in BAT and all major white adipose depots in mice. VEGF expression was increased during white and brown adipocyte differentiation and was regulated in cultured brown adipocytes by the \(PPAR\gamma\) agonist troglitazone and by \(PGC1\alpha\) in BAT in vivo. Systemic VEGF neutralization led to brown adipocyte apoptosis in vivo, loss of mitochondrial cristae and increased mitophagy and was associated with increased inflammation and fibrosis. VEGFR2 was expressed in both brown preadipocytes and adipocytes. Blockade of VEGF signaling using anti-VEGFR2 antibody DC101 increased brown adipocyte apoptosis in vitro. VEGF also functioned as a mitogen and survival factor for brown preadipocytes. VEGF 164 and VEGF 188, isoforms that can bind heparan sulfate proteoglycans, comprise >98% of total VEGF in BAT, subcutaneous and perigonadal fat depots. Embryos that lacked VEGF 164 and 188 displayed abnormal BAT development with fewer brown adipocytes, lower levels of mitochondrial uncoupling protein 1 and Cox IV. These results indicate a direct role for VEGF signaling in brown adipocytes and preadipocytes and suggest the importance of heparan sulfate binding VEGF isoforms in BAT development. Elucidation of the role of VEGF signaling in adipocytes is vital to understanding adipose tissue expansion and activation and may reveal novel therapeutic targets for the activation of brown fat in humans.

Πρόσφατα δεδομένα σε πειραματικά μοντέλα ζώων αλλά και σε ασθενείς καταδεικνύουν την σημαντικότητα της «ποιότητας» της λιποπρωτεΐνης υψηλής πυκνότητας (HDL), σε σχέση με την ποσότητά της, στην προστασία από τη στεφανιαία νόσο. Τα κύρια χαρακτηριστικά της HDL τα οποία καθορίζουν την «ποιότητά» της, είναι η ανάστροφη μεταφορά χοληστερόλης, οι αντιοξειδωτικές και αντιφλεγμονώδεις ιδιότητές της καθώς επίσης και η ικανότητά της να προάγει την παραγωγή ΝΟ στο αρτηριακό επιθήλιο και να ρυθμίζει την ομοιόσταση της γλυκόζης αίματος. Στην συγκεκριμένη ερευνητική εργασία μελετάμε την επίδραση που έχει η ενεργοποίηση του φαιού λιπώδους ιστού, μέσω της έκθεσης φυσιολογικών πειραματικών μοντέλων μυών, σε περιβάλλον χαμηλής θερμοκρασίας, στο λιπιδαιμικό προφίλ, καθώς και στην λειτουργικότητα της HDL. Είναι γνωστό είναι ότι η ενεργοποίηση του φαιού λιπώδους ιστού, λόγω έκθεσης πειραματόζωων σε περιβάλλον χαμηλής θερμοκρασίας, οδηγεί σε θερμογένεση, κάθαρση των τριγλυκεριδίων από το αίμα παχύσαρκων ποντικών, μείωση του βάρους τους, έλεγχο του καταβολισμού των λιποπρωτεϊνών και ομαλοποίηση της ανοχής τους στην γλυκόζη. Έτσι θελήσαμε να επεκτείνουμε την μελέτη μας, στον ενδεχόμενο ρόλο του εγκλιματισμού των πειραματόζωων μετά από μακρόχρονη έκθεσή τους στο ψύχος στους παραπάνω παράγοντες. Για την επίτευξη των πειραματικών μας στόχων δημιουργήθηκαν τρεις ομάδες C57BL/6 μυών εκ των οποίων η μια εκτέθηκε σε περιβάλλον χαμηλής θερμοκρασίας για 2 εβδομάδες (βραχύχρονη έκθεση στο ψύχος), η άλλη ομάδα για 12 εβδομάδες (μακρόχρονη έκθεση στο ψύχος), ενώ η τρίτη αποτέλεσε την ομάδα ελέγχου. Στις δύο ομάδες μυών έγιναν κινητικές μελέτες έκκρισης και κάθαρσης τριγλυκεριδίων. Επίσης απομονώθηκαν από το πλάσμα του αίματος, τα λιποπρωτεϊνικά κλάσματα στα οποία έγιναν αναλύσεις για τον έλεγχο της ποσότητας και της ποιότητας της HDL. Τα αποτελέσματά μας καταδεικνύουν την επαγωγή θερμογένεσης από τον φαιό λιπώδη ιστό και στις δύο πειραματικές ομάδες, με αυτή να είναι πιο έντονη κατά την βραχύχρονη έκθεση στο ψύχος. Παρόλο που η κατανάλωση τροφής στις ομάδες που εκτέθηκαν στο ψύχος αυξήθηκε, το σωματικό τους βάρος παρέμεινε το ίδιο, μιας και η περίσσεια διατροφικών λιπιδίων χρησιμοποιούνταν από τον φαιό λιπώδη ιστό για την παραγωγή θερμότητας παρά για αποθήκευσή τους στον λευκό λιπώδη ιστό. Η εντερική απορρόφηση τριγλυκεριδίων αυξήθηκε σε σχέση με την ομάδα ελέγχου καθώς πιθανά ο οργανισμός χρειαζόταν τα λιπίδια για θερμογένεση, ενώ η ηπατική έκκριση τριγλυκεριδίων μειώθηκε. Η ολική χοληστερόλη των λιποπρωτεϊνικών κλασμάτων, στις πειραματικές ομάδες, έτεινε να βρίσκεται συγκεντρωμένη στα πιο ώριμα κλάσματα, ιδιαίτερα έπειτα από μακρόχρονη έκθεση στο πειραματικό περιβάλλον. Επιπλέον η HDL την 12η εβδομάδα, παρουσίασε καλύτερη αντιοξειδωτική δράση καθώς και διατήρησε αμιγή την ικανότητα να επιτελεί εκροή χοληστερόλης από τα κύτταρα. Τα ευρήματά μας οδηγούν σε δύο ενδιαφέρουσες ανακαλύψεις: 1) Η βραχεία έκθεση στο ψύχος έχει μεγαλύτερη επίδραση στη θερμογένεση και την παραγωγή ATP, απ’ ότι η μακρά έκθεση. 2) Η μακρά έκθεση στο ψύχος οδηγεί σε πιο λειτουργική HDL σε σχέση με την ομάδα βραχείας έκθεσης. Κατά συνέπεια τα αποτελέσματα μας αυτά, καταδεικνύουν εγκλιματισμό των πειραματόζωων στο ψύχος, ο οποίος συνοδεύεται από ποιοτικότερη και πιο λειτουργική HDL. / Recent data indicate the significance of “HDL quality” in atherosclerosis, rather than HDL cholesterol levels in plasma, suggesting that composition as well as functionality of HDL particle, imparts the atheroprotective property of HDL. The main atheroprotective property of HDL is cholesterol efflux in addition to anti-inflammatory and antioxidant properties. HDL has also the ability to promote NO production in the arterial lining and to regulate blood glucose levels. In this study we intend to examine whether brown adipose tissue activation, through low environmental temperature, affects lipid profile and functionality of HDL in C57BL/6 mice. Previous studies have shown that BAT is activated in animals which have been exposed to low temperature environment, resulting in thermogenesis, blood triglycerides clearance, weight reduction, control of lipoproteins catabolism and normalization of glucose tolerance. Thus we intended to study the possible role of acclimatization after long-term exposure to cold temperature regarding the above factors. Therefore we formed three groups of C57BL/6 mice, one of which was exposed to 4oC environment for 2 weeks (short-term exposure), the other one for 12 weeks (long-term exposure) and the last one at 28oC environment (control group). Kinetic studies of triglyceride secretion and clearance where performed, to all groups of animals. Analyses regarding HDL quantity and quality, where performed in lipoprotein fractions isolated from plasma. Our data demonstrate the induction of thermogenesis in BAT in both experimental groups, which appeared to be more intense during the short-term exposure. Although food consumption, in groups exposed to cold, was increased, body weight did not change, as BAT used the excess dietary lipids in order to produce heat rather than storing them in white adipose tissue. Intestinal absorption of triglycerides was increased compared to the control group, possibly because lipids are needed due thermogenesis. Total cholesterol in HDL fractions, tended to be concentrated in more mature fragments, especially after long-term exposure. In addition HDL, during long-term exposure, showed to have more effective antioxidant potential and cholesterol efflux, compared to the group exposed to cold temperature for 2 weeks. Our observations lead to two interesting findings: 1) Short-term exposure to cold has greater effect on thermogenesis and ATP production rather than long-term exposure. 2) After long-term exposure to cold, HDL appears to be more functional compared to the short-term exposure group. Therefore our results indicate acclimation of the animals to low temperature environment, followed by a qualitative HDL.

Human beings have been exposed to different cold conditions throughout time, and have through cold acclimation developed mechanisms to survive in these conditions. Cold acclimation can be elicited through exposure to natural cold climates, or artificially induced in a laboratory to study the body’s response to repeated cold exposures. Several studies looking at the effects of cold acclimation in humans have been conducted during the last 50 years, and have reported that cold acclimation can lead to a change in skin and core temperature, heat production and shivering. An accurate quantification of shivering thermogenesis (ST) during cold acclimation has not been done before, and most previous measurements of shivering during cold acclimation have been inaccurate and inadequate. In this study a Liquid Condition Suits (LCS) was used to elicit cold acclimation (10°C, 2hr daily, for 4 weeks) while an accurate measurement of the effect of cold acclimation on changes in muscle activity was conducted. In CHAPTER 2, results showed that four weeks of cold acclimation at 10°C did not change skin and core temperature, heat production or ST. The effects on shivering pattern and fuel selection were also analysed, but no effects of cold acclimation could be observed. These measurements were a part of a larger study, in which the effects of cold acclimation on changes in BAT were the main outcome measures. These data showed that an increase in BAT volume (45%) and activity (120%) were the only observed effects of cold acclimation. In CHAPTER 3, we set out to assess if changes in shivering from pre to post cold acclimation are associated with changes in BAT volume, and if the amount of BAT a participant possesses prior to cold acclimation can be used to predict changes in shivering intensity during cold acclimation. The interindividual variability in changes in thermal responses, heat production, shivering and BAT volume occurring between subjects during four weeks of cold acclimation was also addressed in this section.

Submitted by CARMEM PERES VALGAS DA SILVA (carmempvs@hotmail.com) on 2018-06-05T14:40:45Z No. of bitstreams: 1 TESE FINAL jun 2018 (1).docx.pdf: 3453585 bytes, checksum: 2f207fc1ad5f86e08a3cb3497d63daf9 (MD5) / Approved for entry into archive by Ana Paula Santulo Custódio de Medeiros null (asantulo@rc.unesp.br) on 2018-06-05T16:33:57Z (GMT) No. of bitstreams: 1 silva_cpv_dr_rcla.pdf: 3391182 bytes, checksum: 8bfcf2d76191e2dfdab26a4cc2057921 (MD5) / Made available in DSpace on 2018-06-05T16:33:57Z (GMT). No. of bitstreams: 1 silva_cpv_dr_rcla.pdf: 3391182 bytes, checksum: 8bfcf2d76191e2dfdab26a4cc2057921 (MD5) Previous issue date: 2018-04-12 / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O sistema nervoso simpático desempenha importante papel no sistema cardiovascular e no metabolismo, sendo que os receptores adrenérgicos do subtipo β3 (β3 –AR) estão amplamente distribuídos em diferentes células, encontrando-se especialmente no tecido cardíaco, musculatura lisa e tecido adiposo. Esses receptores estão relacionados a diversas ações fisiológicas, como relaxamento da musculatura lisa vascular e não vascular, lipólise do tecido adiposo branco e termogênese do tecido adiposo marrom. Por outro lado, a importância dos β3 –AR no tecido adiposo perivascular (PVAT) e regulação do tono vascular não é conhecida. O mirabegron (YM-178) é um agonista β₃-AR seletivo, atualmente desenvolvido por Astellas Pharma Inc., e aprovado desde 2012 pelos EUA, para tratamento de bexiga hiperativa, considerando a ação dos receptores β₃-AR no relaxamento do músculo liso. Contudo não existe nenhum estudo envolvendo o mirabegron nos parâmetros cardiometabólicos plasmáticos e no metabolismo do tecido adiposo e em modelos de obesidade. Objetivos: Analisar os efeitos do tratamento de Mirabegron (Mira), um agonista β₃-AR seletivo, por duas semanas em parâmetros cardiometabólicos e reatividade vascular na presença ou ausência de PVAT de camundongos obesos. Métodos: Camundongos C57BL/6J foram divididos em grupos controle veículo (CTR), controle tratado com Mira (CTR+MIRA), obeso alimentado com dieta hiperlipídica (HFD 12 semanas) tratado com veículo (OB) e obeso tratado com Mira (OB+MIRA). Mira foi administrado por duas semanas (10mg/kg) na forma de gavagem. Curvas concentração-resposta à acetilcolina (ACh 100pM a 30 μM) e serotonina (5HT 1nM a 30μM) foram realizadas em anéis de aorta PVAT+, PVAT-. Foram medidos a temperatura corporal e gasto energético; bem como pressão arterial, frequência cardíaca e tolerçancia ao esforço, além de análises bioquímicas de perfil lipídico, glicemia de jejum, insulina sérica, Glicerol, AGL, TBARS, leptina e TNF-α. Foram calculados o índice HOMA e o índice aterogênico. A histologia hepática e dos depósitos de tecido adiposo foi realizada pela coloração de hematoxilina e eosina. Resultados: O tratamento com Mira (OB+ mira) por duas semanas aumentou o gasto energético (17%) sem alterar a temperatura corporal, redução do peso corporal (7%), gordura epididmal (25%), colesterol total (17%), LDL-C (55%), índice aterogênico (45%), TBARS (20%), TNF-α (48%), Glicerol e AGL (40%), insulina (67%), e índice HOMA (65%) quando comparados ao grupo OB. Os níveis elevados de leptina (1000%) no grupo OB não foram alterados pelo Mira. Como esperado os depósitos de lipídios hepáticos, do TAM e PVAT foram aumentado com a dieta HFD e Mira reverteu parcialmente essa alteração, observados na histologia. Observamos modificação nas gotículas lipídicas tecido adiposo inguinal dos grupos CTR+MIRA e OB+MIRA. Anéis com PVAT+ apresentaram aumento na resposta máxima (Emax) à 5HT (%KCl) em anéis de aorta de OB+MIRA (109±10) comparado aos outros grupos (CTR: 69±9; CTR+MIRA: 77±11; OB: 65±9). Conclusão: Em camundongos obesos, o tratamento com Mira por duas semanas resultou em modificação dos tecidos adiposos marrom e inguinal, aumento do gasto energético, melhora de importantes parâmetros bioquímicos como o perfil lipídico e glicêmico, além de reduzir o marcador de estresse oxidativo TBARS, sem contudo alterar a tolerância ao esforço, pressão arterial e frequência cardíaca. Entretanto, em anéis de aorta PVAT+, o tratamento com Mira resultou em aumento da Emax para 5-HT. / Sympathetic nervous system plays an important role in the cardiovascular system and metabolism. In particular, the β3 subtype adrenergic receptors (β3-AR) are widely distributed in different cells, especially in cardiac tissue, smooth muscle and adipose tissue. These receptors are related to several physiological actions, such as vascular and non-vascular smooth muscle relaxation, lipolysis and thermogenesis. In addition, β3-AR participates in the increasing of the number of mitochondria at specific sites of white adipose tissue, called browning of white adipose tissue. Finally, it is not known whether these receptor subtypes may play a role in regulation of the perivascular adipose tissue (PVAT) vasomotricity. Despite these important actions involving these receptor population, studies involving selective β3-AR agonists on obesity state are scarce. Mirabegron (YM-178) is a selective β₃-AR agonist, currently developed by Astellas Pharma Inc. and approved since 2012 by the USA for overactive bladder treatment, considering the action of β₃-AR receptors on smooth muscle relaxation. Intrestingly, there is no study involving the effects of mirabegron on cardiometabolic parameters, adipose tissue metabolism in obesity models. Objectives: To examine the effects of Mirabegron (Mira) treatment, for two weeks on cardiometabolic parameters and vascular reactivity in the presence or in the absence of PVAT in obese mice. Methods: C57BL / 6J mice were divided into vehicle control (CTR), Mira treated (CTR + MIRA), obese mice fed with hyperlipid diet (HFD 12 weeks) treated with vehicle (OB) and obese treated with Mira (OB + MIRA). Mira was given for two weeks (10mg / kg) as a gavage. Concentration-response curves to Acetylcholine (ACh 100pM at 30μM) and serotonin (5HT 1nM at 30μM) were obtained in PVAT + aorta rings, PVAT-. Body temperature and energy expenditure were measured as well as biochemical parameters: lipid profile, fasting glycemia, serum insulin, glycerol, FFA, TBARS, leptin and TNF-α. HOMA index and atherogenic index were calculated. Hepatic and adipose tissue histology was performed by staining hematoxylin and eosin. Results: Treatment with Mira for two weeks increased energy expenditure (17%), without alteration in body temperature. Reduction is body weight (7%), epididymal fat (25%), total cholesterol (17%), LDL-C (55%), atherogenic index (45%), TBARS %), TNF-α (48%), Glycerol and AGL (40%), insulin (67%) and HOMA index (65%) was seen in OB + MIRA when compared with OB group. The increase in leptin levels (1000%) in OB group was not modified by Mira. In contrast, the increased lipid deposits in hepatic, TAM and PVAT, observed in histology, was partially reversed by Mira. We observed changes in lipid droplets in the inguinal adipose tissue of CTR+MIRA and OB+MIRA groups. Rings with PVAT + showed an increase in the Emax to 5HT (% KCl) in aortic rings of OB + MIRA (109 ± 10) compared with the other groups (CTR: 69 ± 9; 65 ± 9; CTR+MIRA: 77±11; OB: 65±9). Conclusion: In obese mice, treatment with Mira for two weeks resulted in modification of brown and inguinal adipose tissues, an increase in energy expenditure, improvement of biochemical parameters such as lipid profile and glycemia, without any modification in the tolerance to exercise effort, blood pressure and heart rate. On the other hand, in PVAT + aortic rings, Mira treatment resulted in increased in Emax to 5HT.

Abstract The use of regulatory and obligatory heat production mechanisms were studied in juvenile birds during the development of endothermy. The development of shivering thermogenesis was studied in the pectoral and gastrocnemius muscles of the altricial domestic pigeon and in three precocial galliforms (Japanese quail, grey partridge and domestic fowl). The development of shivering was the determinant for the beginning of endothermy. Homeothermy also necessitated avoidance of excess heat loss by insulation and behavioural thermoregulation. In the precocial species, shivering thermogenesis was present in the leg muscles of the youngest age groups (1-2 d) studied. Breast muscles contributed shivering from the second post-hatching week. In the altricial pigeons, significant thermogenesis was apparent later than in the precocials, at the age of 6 d. In contrast to the precocials, the pectoral muscles of the altricials were the most significant heat production tissues. In newly-hatched partridges and pigeons, incipient shivering did not result in significant heat production. The ability to produce heat in cold by putative nonshivering thermogenesis was studied in Japanese quail chicks and domestic ducklings. In both species, three-week cold acclimation resulted in morphometric and physiological changes, but there was no clear evidence of nonshivering thermogenesis. The lack of NST was evident because an increase in shivering amplitude at least in one of the muscles studied paralleled an increase in oxygen consumption. Consequently, shivering thermogenesis was probably the only mode of regulatory heat production. The amplitudes of shivering EMGs measured during cold exposure were dependent on the coexistence of postprandial thermogenesis or exercise. Japanese quail chicks were able to substitute shivering thermogenesis partially with postprandial heat production when nourished. Bipedal exercise both inhibited shivering in pectorals directly via inhibitory neural circuits and stimulated it indirectly via decreased body temperature. Because of increased heat loss, exercise was not used as a substitute for shivering. Shivering is a flexible mode of thermogenesis and its magnitude can be adjusted according to the magnitude of obligatory thermogenesis. The adjustment works towards energy saving by avoidance of the summation of different modes of heat production. The prerequisite for successful adjustment of shivering is adequate insulation, whose role in preventing excessive heat loss is pronounced during exercise. It is concluded that the energetics of posthatching thermoregulation includes the potential for optimizations in energy use in order to avoid dissipation of waste energy as heat.

Human beings have been exposed to different cold conditions throughout time, and have through cold acclimation developed mechanisms to survive in these conditions. Cold acclimation can be elicited through exposure to natural cold climates, or artificially induced in a laboratory to study the body’s response to repeated cold exposures. Several studies looking at the effects of cold acclimation in humans have been conducted during the last 50 years, and have reported that cold acclimation can lead to a change in skin and core temperature, heat production and shivering. An accurate quantification of shivering thermogenesis (ST) during cold acclimation has not been done before, and most previous measurements of shivering during cold acclimation have been inaccurate and inadequate. In this study a Liquid Condition Suits (LCS) was used to elicit cold acclimation (10°C, 2hr daily, for 4 weeks) while an accurate measurement of the effect of cold acclimation on changes in muscle activity was conducted. In CHAPTER 2, results showed that four weeks of cold acclimation at 10°C did not change skin and core temperature, heat production or ST. The effects on shivering pattern and fuel selection were also analysed, but no effects of cold acclimation could be observed. These measurements were a part of a larger study, in which the effects of cold acclimation on changes in BAT were the main outcome measures. These data showed that an increase in BAT volume (45%) and activity (120%) were the only observed effects of cold acclimation. In CHAPTER 3, we set out to assess if changes in shivering from pre to post cold acclimation are associated with changes in BAT volume, and if the amount of BAT a participant possesses prior to cold acclimation can be used to predict changes in shivering intensity during cold acclimation. The interindividual variability in changes in thermal responses, heat production, shivering and BAT volume occurring between subjects during four weeks of cold acclimation was also addressed in this section.

High altitude is characterized by chronically low ambient temperatures and oxygen. To survive, highland native deer mice (Peromyscus maniculatus) are capable of high rates of prolonged thermogenesis due to elevated aerobic capacity (V̇O2max) in hypoxia. Deer mice primarily use fats to fuel their high metabolic rates for heat production. Carnitine palmitoyl-transferase 1 (CPT-1) is a rate-limiting step in mitochondrial fat oxidation, and a reduction in CPT-I sensitivity for its substrate L-carnitine is associated with a reduction in muscle fat use during high intensity exercise in mammals. Sensitivity of mitochondrial metabolism to ADP also changes with exercise. It is currently unknown whether similar mechanisms underpin regulation of fuel use during shivering, but I predicted that sensitivities to ADP and L-carnitine would be greater in highlanders than lowlanders and increase with acclimation. To address this question, I examined mitochondrial sensitivity to substrates involved in the fat oxidation pathway in low- and high- altitude deer mice born and raised in common laboratory conditions. Mice were also acclimated to high altitude condition of cold hypoxia to examine if the plasticity of these traits were affected by altitude ancestry. Consistent with previous findings, both high and lowland mice increased their cold-induced V̇O2max following cold hypoxia acclimation and rely primarily on lipids to fuel thermogenesis. High- and low-altitude deer mice responded differently to chronic cold hypoxia with highlanders showing a ~7-fold greater ADP sensitivity than lowlanders following acclimation. In contrast to the expected outcome, highlander deer mice tended to have a reduced sensitivity to L-carnitine compared to lowlanders that approached statistical significance. Neither sensitivity to palmitoylcarnitine sensitivity nor mitochondrial expression of FAT/CD36, thought to aid in mitochondrial fat delivery, showed differences between population or changes with acclimation, indicating that limitations to lipid oxidation during shivering likely occur at, or upstream of, CPT-I in the deer mouse. / Thesis / Master of Science (MSc) / Some animals can survive extremely harsh climates, such as high altitude. High altitude is characterized by unremitting cold and thin air, and these challenges can constrain aerobic activities in mammals. The North American deer mouse can thrive at high altitude by actively generating large amounts of body heat in a process known as thermogenesis. The deer mouse relies primarily on fats as fuel to support thermogenesis, but the cellular mechanisms that regulate the use of lipids to power thermogenesis remain unclear. To address this question, I induced shivering in deer mice from both high- and low-altitude populations that I exposed to simulated high- or low-altitude conditions. I then examined the effects of these treatments on the ability of shivering muscle to consume oxygen and fuel for thermogenesis. My thesis contributes to the current understanding of how mammals manage their energy supply to survive in a challenging environment.

No description available.

No description available.