No description available.

No description available.

No description available.

Uncoupling proteins (UCPs) constitute a highly conserved subset of mitochondrial solute carriers. Discovered in small rodents in the early 1970’s, UCPs and their homologs have since been found in nematodes, plants, birds, and, most recently, in significant depots within humans (Krauss et al. 2005, Van marken Lichtenbelt 2009). Following activation by long chain fatty acids (LCFA, e.g. oleic acid) and reactive oxygen species (ROS, e.g. 4-hydroxynonenal (4HNE)), UCPs form a proton channel within the inner mitochondrial membrane and permit the influx of hydrogen ions from the inter membrane space into the mitochondrial matrix. UCPs effectively uncouple oxidative phosphorylation (OX-PHOS) from ATP generation, resulting in increasing oxygen consumption and dissipating the chemical energy in the form of heat. Found primarily in brown adipose tissue (BAT) of small hibernating mammals, the canonical role of uncoupling protein 1 (UCP1) in mammalian adaptive thermogenesis has been thoroughly studied. However, UCP1 is not the only member of the uncoupling family found within BAT. Also playing a key role in this tissue is uncoupling protein 3 (UCP3), which is a close homolog to UCP1. However, in spite of the fact that UCP3 shares more than 50% amino acid homology and tissue localization with UCP1, the true function UCP3 is very poorly elucidated. Part of the difficulty in determining this function lies in the expression levels of the UCP3 protein, which are hundreds of folds less than UCP1 in this tissue. In addition, their homologous structure makes teasing apart UCP3-specific phenomena from UCP1-mediated mechanisms very difficult using conventional techniques in cell and molecular biology. While UCP1 is almost exclusively found in BAT, UCP3 is expressed primarily in skeletal muscle (SKM), which lacks UCP1 completely (Krauss et al. 2005). Because UCP3 is so enriched in SKM, many studies have focused on its role in that tissue and have then tried to transpose these functions into BAT. As a result, UCP3 has been implicated in facilitating numerous biological processes, including non-adaptive facultative thermogenesis, affecting SKM oxidative capacity by modulating LCFA export, and ameliorating elevated levels of ROS-mediated stress within the tissue via glutathionine (GSH) interacting moieties. Ultimately, however, little consensus exists on the function of UCP3 within SKM, and subsequently, even less is known about its purpose in BAT. Previous data has shown that murine UCP1 has the capacity to bind to itself and form homo-tetramers when expressed in vitro in recombinant E. coli (Hoang T. et al. 2013). Here we show that UCP1 interacts with UCP3 in BAT in vivo, supporting Hoang’s research above by showing that UCP1 has the capacity to not only homodimerize but potentially oligomerize with other UCP homologs. While many groups using UCP3-null mice have reported no gross changes in physiologic responses, data previously published in the lab showed that mice lacking UCP3 were protected from potentially fatal hyperthermic effects when administered sympathomimetic agents such as 3,4-Methylenedioxymethamphetamine (MDMA), methamphetamine (METH), lipopolysaccharide (LPS), or norepinephrine (NE) (Mills et al. 2003, Kenaston et al. 2010). This implies that UCP3 plays an intimate role in sympathetic nervous system (SNS) mediated thermogenesis. Based upon the foregoing, the primary goal of the research discussed in this thesis was to elucidate the functions of UCP3 within BAT. In this study, we recapitulated results seen by other students in this lab: that global UCP3-null mice do indeed exhibit a blunted thermogenic response when treated with sympathomimetic agonists. In addition, despite the near-ubiquitous expression of UCP2 throughout the mammalian organism, this UCP is not involved in SNS-mediated thermogenesis (Arsenijevic et al. 2000). Our data shows that UCP3 is vital to the catecholamine-mediated thermogenic responses following sympathomimetic drug administration. When challenged by METH, UCP3-null mice were able to respond, albeit with a blunted increase in body temperature. Furthermore, when challenged by NE, a key neurotransmitter involved in mediating the responses initiated by the SNS following METH exposure, UCP3-null mice were able to mount half the hyperthermic response seen in WT littermates. However, UCP1/UCP3 double-null animals exhibited an almost four-fold hypothermic effect compared to WT littermates when challenged with NE. In addition, UCP1/UCP3 double-null mice were unable to restore body temperatures back to baseline values, an effect seen in all the other genotypes. This implies that UCP3 plays an important role in restoring body temperatures to physiological norms. Therefore, while the mechanism underlying the decreased responsiveness to NE remains unclear, it is clear that whether localized to SKM or BAT, UCP3 is a major player in the mammalian response to SNS-mediated thermogenesis and global thermoregulation. / text

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Humans are known as homeothermic endotherms. To ensure thermic balance at rest when exposed to cold, they dispose of two main thermogenic processes: shivering thermogenesis (ST) and non-shivering thermogenesis (NST). ST consists of involuntary muscle contractions and NST represents the component of Hprod that is not ST. While ST is difficult to tolerate, it is not yet known which nutrients and how much are required to stimulate NST and lower ST in the cold. Whether or not food caloric intake has an impact on the relative contribution of ST and NST to total Hprod remains to be determined. Therefore, the purpose of this thesis was 1) to quantify the effects of ingesting two quantities (1507 vs 3015 kJ) of same relative compositions on cold-induced whole-body Hprod and 2) to establish the effects of these two quantities of food on the relative contribution of ST and NST to total Hprod. Five healthy male participants were exposed to a 3h mild cold, using a liquid conditioned suit with water flowing at 15°C (COLD) or 33 °C (CON) for a total of 4 trials. Thermal, metabolic and shivering responses were measured at baseline, before and after shake ingestion. Results demonstrated that Hprod and ST intensity increased in the cold, while no significant differences were found between the ingested shakes at two different caloric equivalents. In addition, ST intensity did not change, which confirmed that NST remained the same between the two conditions. Thus, knowing that the caloric intake will not maximize the thermogenic effects in the cold (i.e. improve the comfort of the individual), is it more advantageous to bring food or additional clothing, for any activity? Clearly, more research on the exact pathways of each processes in the cold with food consumption needs to be made. To that extent, the investigation of the effect of food quality on changes in the thermogenic processes during cold exposure strikes us as a fascinating area for future research.

No description available.

Maintenance of body temperature is one of the first and most important physiological processes that must be initiated after birth. Failure to sustain homeothermy leads to hypothermia and death. Indeed, in sheep, 40% of non-predator lamb deaths are attributed to cold and cold-related causes. Brown adipose tissue (BAT) is an essential mediator of thermogenesis in many species and is responsible for 50% of the heat generated in the newborn lamb despite comprising only 2% of body weight. Previously, we found that maternal arginine supplementation increased fetal peri-renal BAT by 62%. This observation led us to test the hypothesis that increased the amount of fetal BAT will enhance neonatal thermogenesis at birth and thus combat the effects of cold stress. Thirty-one multiparous Suffolk ewes gestating singletons and twins were assigned to receive either intravenous injections of L-arginine (27 mg/kg bodyweight; n=17) or sterile saline (n=14) three times daily from Day 75 to Day 125 of gestation (term=147). Following parturition lambs were removed from their dams, placed in a thermoneutral environment, and fed artificial colostrum on a per body weight basis. At 4 hours of age, lambs were cold challenged at 0 degrees C for 2 hours. Rectal temperatures were recorded at 15 minute intervals. At 6 hours of age all singletons and one lamb of each twin pair was sacrificed. The remaining twin lamb was challenged again at 22 hours of age for an additional 2 hours prior to necropsy. Rectal temperature was greater for the duration of both cold challenges in lambs from arginine-treated ewes than lambs from saline-treated ewes (P<0.050). Interestingly, at time of necropsy, BAT weight did not differ (P>0.10) between treatments. UCP1 mRNA levels were not affected by treatment or age (P>0.10). However, TEK, PPARGC1A, NRF1, NRF2, PPARG, ADRB3, ARG2, RPS6KA1, EIF4EBP1, ODC1 were not affected by treatment (P>0.10) but were upregulated (P<0.05) by age; being greater at 24 hours of age versus 6 hours of age. Results indicate that maternal arginine treatment results in increased neonatal thermogenesis after birth. Although the underlying mechanisms remain to be elucidated, the data reported herein represent the first step in improving neonatal survival in response to cold.

High altitude is one of the most extreme environments inhabited by endotherms, where extreme cold temperatures and low O2 availability (hypoxia) can constrain aerobic heat production (thermogenesis) to maintain body temperature (Tb). Recent findings suggest that the gut microbiome contributes to whole-body thermogenesis, but the significance of this mechanism for coping in cold environments is unknown. We examined whether the gut microbiome contributes to adaptive variation in thermogenic performance in deer mice at high altitude. Mice from populations native to high altitude and low altitude were born and raised in common conditions. Adults from both populations were acclimated to warm (25C) normoxia or cold (5C) hypoxia (~12 kPa O2 for 6 weeks) in a full factorial design, and a subset of mice in each group were treated with antibiotics to deplete the gut microbiome. Thermogenic endurance was then measured as the duration that Tb and metabolism could be maintained during acute cold challenge. In lowlanders, antibiotics had only modest effects on thermogenic endurance. In highlanders, in stark contrast, antibiotic treatment led to pronounced reductions in thermogenic endurance in both environments. These effects could not be explained by impairments in aerobic heat production by host thermogenic tissues, because antibiotic treatment had no effects on cold-induced increases in O2 consumption or UCP1 content of brown adipose tissue. These results suggest that the gut microbiome plays an increased role in thermogenesis in high-altitude mice. Thermogenic performance contributes to fitness at high altitude, suggesting that changes in host-microbe interactions contribute to high-altitude adaptation. / Thesis / Master of Science (MSc) / High altitude is one of the most extreme environments inhabited by endotherms. Recent findings suggest that the gut microbiome contributes to thermogenesis, but the significance of this mechanism for coping in cold environments is unknown. We examined whether the gut microbiome contributes to adaptive variation in thermogenic performance in deer mice at high altitude. Adult mice from low and high-altitude populations were acclimated to warm normoxia or cold hypoxia, and a subset of mice in each group were treated with antibiotics to deplete the gut microbiome. Thermogenic endurance was measured as the duration that Tb and metabolism could be maintained during acute cold challenge. In lowlanders, antibiotics had modest effects on thermogenic endurance but in highlanders, it led to pronounced reductions in thermogenic endurance in both environments. These results show that thermogenic performance contributes to fitness at high altitude, suggesting that changes in host-microbe interactions contribute to high-altitude adaptation.

It has been demonstrated that EGCG and caffeine, naturally present in green tea, have thermogenic properties in thermoneutral conditions. The purpose of this study was to quantify the effect of the combined ingestion of EGCG/caffeine on thermogenic responses during a 3h mild cold exposure. Eight healthy males (22± 1 y) were exposed in a randomized, cross over, single blinded fashion to the cold (liquid conditioned suit perfused with 15°C water), after ingesting either a placebo (CON) or an extract of 1600mg of EGCG and 600mg of caffeine (EXP). Thermic, metabolic and electromyographic measurements were monitored at baseline and during cold exposure. After 180min of cold exposure, shivering intensity was significantly reduced by ~32% in EXP condition compared to CON. Area under the curve calculations for total shivering intensity was also reduced by ~21% in EXP (457±99 %MVC.min) compared to CON (361±81 %MVC.min; p=0.007). In contrast, the total area under curve of VO2 was ~25% higher in EXP (33.3±5.5 L O2) compared to CON (25.3±5.1 L O2; p=0.03). Total Heat production (Hprod) also increased by about 11% in the EXP condition (1535±112 kJ) compared to control (1372 ±106 kJ; p=0.002). The decrease in shivering activity combined with an increase in VO2 and Hprod, following the ingestion of EGCG and caffeine in the cold, indicates that nonshivering thermogenesis pathways can be significantly stimulated in adult humans.