Synergic effect of overweight and cold on uncoupling proteins expression,a role of alpha(2)/beta(3) adrenergic receptor balance?

The effect of cold exposure, being overweight and their interaction was investigated on the response of uncoupling proteins UCP1, UCP2 and UCP3 and the alpha(2)/beta(3) adrenergic receptor (AR) balance in brown adipose tissue (BAT), as well as the involvement of leptin gene expression in white adipose tissues, in control and overweight male rats of the dietary obesity model known as the post-cafeteria model. UCP1, UCP2 and UCP3 mRNAs were up-regulated by cold, with a synergic effect of cold exposure and being overweight on UCP1 mRNA levels (with the related UCP1 protein response), and with UCP2 mRNA showing a parallel response. Furthermore, the BAT alpha(2)/beta(3) AR ratio was diminished in overweight rats. The results suggest that the UCP1-dependent thermogenic capacity in BAT of post-cafeteria overweight rats has a more sensitive response to cold exposure and that UCP2 and UCP3 could be somehow involved in the thermogenic response but differentially regulated. Moreover, the diminished alpha(2)/beta(3) AR ratio in BAT could be one of the factors involved in the more sensitive response of overweight rats to cold in terms of BAT thermogenesis-related parameters.     

Leanness of Lou/C rats does not require higher thermogenic capacity of brown adipose tissue

Lou/C rats, an inbred strain of Wistar origin, remain lean throughout life and therefore represent a remarkable model of obesity resistance. To date, the exact mechanisms responsible for the leanness of Lou/C rats remain unknown. The aim of the present study was to investigate whether the leanness of Lou/C rats relies on increased thermogenic capacities in brown adipose tissue (BAT).Results showed that although daily energy expenditure was higher in Lou/C than in Wistar rats, BAT thermogenic capacity was not enhanced in Lou/C rats kept at thermoneutrality as demonstrated by reduced thermogenic response to norepinephrine in vivo, similar oxidative activity of BAT isolated mitochondria in vitro, similar levels of UCP1 mRNA and lower abundance of UCP1 protein in interscapular BAT depots. Relative abundance of β₃-adrenergic receptor mRNA was lower in Lou/C BAT while that of GLUT4, FABP or CPT1 was not altered. Activity-related energy expenditure was however considerably increased at thermoneutrality as Lou/C rats demonstrated an impressively high spontaneous running activity in voluntary running wheels. Prolonged cold-exposure (4 °C) depressed the spontaneous running activity of Lou/C rats while BAT thermogenic capacity was increased as reflected by rises in BAT mass, oxidative activity and UCP1 expression.It is concluded that the leanness of Lou/C rats cannot be ascribed to higher thermogenic capacity of brown fat but rather to, at least in part, increased locomotor activity. BAT is not deficient in this rat strain as it can be stimulated by cold exposure when locomotor activity is reduced suggesting some substitution between these thermogenic processes.     

Differential down-regulation of β3-adrenergic receptor mRNA and signal transduction by cold exposure in brown adipose tissue of young and senescent rats

 Diminished β₃ adrenergic-stimulated thermogenesis in brown adipose tissue (BAT) in senescent rats is restored by a short period of cold exposure. To investigate if the mechanism of this restoration involves preferential up-regulation of β₃-adrenergic signal transduction in senescent compared with young rats, we examined the steady state levels of β₃-adrenergic receptor messenger ribonucleic acid (β₃AR mRNA) and the dose/response relationship for the activation of adenylyl cyclase by the selective β₃-adrenergic agonist, BRL 37344 in BAT membranes from young and senescent rats with or without 2 days cold exposure at 8 °C. β₃AR mRNA declined by 30% with age and by 60% with cold exposure in young rats. In contrast, cold exposure did not down-regulate β₃AR mRNA in senescent rats. BRL 37344-stimulated adenylyl cyclase activity was 38% less in senescent rats and was desensitized by cold exposure decreasing maximum stimulation by 78% and increasing, the dissociation constant 2.5-fold. Cold exposure decreased BRL 37344-stimulated adenylyl cyclase activity to a lesser extent in senescent rats (50%), such that the activity was similar in cold-exposed young and old rats. These data indicate that although cold exposure did not, as we hypothesized, up-regulate β₃-adrenergic signal transduction in the senescent rats, cold exposure preferentially down-regulated β₃AR mRNA and β₃AR-mediated adenylyl cyclase in the young compared with senescent rats. Received: 22 July 1998 / Received after revision: 9 October 1998 / Accepted: 13 October 1998     

Brown adipose tissue response to cafeteria diet-feeding involves induction of the UCP2 gene and is impaired in female rats as compared to males

Noradrenaline-dependent brown adipose tissue (BAT) thermogenesis is activated by the cold and excess energy intake, largely depends on the activity of the uncoupling protein 1 (UCP1), and is mediated mainly through the beta3-adrenoceptor (beta3-AR). We investigated the expression of ucp2, a gene that encodes a putative UCP1-like uncoupling protein, along with that of ucp1 and beta3-ar, in the interscapular BAT (IBAT) of male and female rats chronically fed a cafeteria diet. After 3 months on this diet, male rats attained a 34% excess body mass and showed IBAT hypertrophy and increased IBAT thermogenic potential, in terms of both UCP1 and UCP2 mRNA expression (both by 1.6-fold), UCP1 protein expression (by 1.75-fold) and GDP binding to IBAT mitochondria (by 2.2-fold); female rats attained a larger excess body weight (50%) and their IBAT, although hypertrophied, showed no signs of increased thermogenic potential per gram of tissue. Interestingly, the IBAT of female rats was already activated compared to males. Treatment of mouse brown adipocytes in primary culture with noradrenaline also triggered a dose-dependent increase of the levels of UCP1 mRNA and UCP2 mRNA. Retroregulatory down-regulation of the beta3-AR mRNA levels was found in the two models used. The results support a physiological role for UCP2, along with UCP1, in rodent BAT thermogenesis.     

Repeated immobilization stress increases uncoupling protein 1 expression and activity in Wistar rats

Repeat immobilization-stressed rats are leaner and have improved cold tolerance due to enhancement of brown adipose tissue (BAT) thermogenesis. This process likely involves stress-induced sympathetic nervous system activation and adrenocortical hormone release, which dynamically enhances and suppresses uncoupling protein 1 (UCP1) function, respectively. To investigate whether repeated immobilization influences UCP1 thermogenic properties, we assessed UCP1 mRNA, protein expression, and activity (GDP binding) in BAT from immobilization-naive or repeatedly immobilized rats (3 h daily for 4 weeks) and sham operated or adrenalectomized (ADX) rats. UCP1 properties were assessed before (basal) and after exposure to 3 h of acute immobilization. Basal levels of GDP binding and UCP1 expression was significantly increased (140 and 140%) in the repeated immobilized group. Acute immobilization increased GDP binding in both naive (180%) and repeated immobilized groups (220%) without changing UCP1 expression. In ADX rats, basal GDP binding and UCP1 gene expression significantly increased (140 and 110%), and acute immobilization induced further increase. These data demonstrate that repeated immobilization resulted in enhanced UCP1 function, suggesting that enhanced BAT thermogenesis contributes to lower body weight gain through excess energy loss and an improved ability to maintain body temperature during cold exposure.     

Blood flow to the brown adipose tissue of conscious young rabbits during hypoxia in cold and warm conditions

 Brown adipose tissue (BAT) non-shivering thermogenesis is stimulated by cold temperature and depressed by hypoxia. We investigated the extent to which changes in metabolic rate during cold and hypoxia, singly or combined, were accompanied by changes in BAT perfusion. One-month-old rabbits were instrumented for measurements of regional blood flow by the coloured microsphere technique. One group of rabbits was tested in warm (24 °C, n=17), and the other in cold (13 °C, n=9) conditions, first in normoxia (inspired oxygen concentration FIO₂ about 21%, arterial oxygen saturation S _aO₂ approximately 88%) followed by hypoxia (FIO₂ approximately 10%, S _aO₂ approximately 54%). In warm conditions, oxygen consumption (V·O₂, measured by an open-flow method) averaged 22 ml·kg^–1·min^–1 (STPD), and BAT blood flow 98 ml·100g^–1·min^–1. In hypoxia, V·O₂ dropped on average to 87%, whereas BAT flow dropped to 43% of the normoxic values. In the cold during normoxia, V·O₂ averaged 31 ml·kg^–1·min^–1 (STPD), and BAT blood flow was 155 ml·100g^–1·min^–1. In cold and hypoxia V·O₂ dropped to 19 ml·kg^–1·min^–1 (STPD) (i.e. 60% of the normoxic value), whereas BAT blood flow was not altered significantly (148 ml·100g^–1·min^–1). Hence, BAT blood flow decreased in hypoxia in absence of cold stimuli, whereas it remained high when hypoxia occurred during cold, despite the major drop in V·O₂. We conclude that cold is more important than hypoxia in determining BAT perfusion, and that changes in BAT blood flow are not a mechanism for the hypoxic control of V·O₂. Received: 24 June 1998 / Received after revision: 21 September 1998 / Accepted: 29 September 1998     

Sexual dimorphism in the adrenergic control of rat brown adipose tissue response to overfeeding

Gender-related differences in the brown adipose tissue (BAT) response to overfeeding rats on a cafeteria diet were studied by assessing the balance between the expression of beta-adrenoceptors (beta1-, beta2-, beta3-AR) and alpha2A-AR and their relation to the expression of uncoupling proteins (UCP1, UCP2, UCP3). Cafeteria diet feeding for 15 days, which involved a similar degree of hyperphagia in both sexes, led to a greater body weight excess in females than in males and a lower activation of thermogenesis. Gender-related differences were found for different adrenoceptor expression and protein levels, which might explain, in part, sex differences in the thermogenic parameters. The lower expression of alpha2A-AR in females than in males could be responsible for the higher expression of UCP1 and thermogenic capacity under non-hyperphagic conditions. However, in a situation of high adrenergic stimulation--as occurs with overfeeding--as there is a preferential recruitment of the beta3-AR by noradrenaline compared with other adrenergic receptors, the higher levels of beta3-AR in males rats than in females could be responsible for the greater thermogenic capacity and the lesser weight gain in males. Thus, the alpha2/beta3 balance in BAT could be a key in the thermogenic control.     

Effects of dehydration and rehydration on body temperatures in the black Bedouin goat

 The temperatures of the arterial blood and the brain in black Bedouin goats were measured continuously by miniature data loggers. The animals were either euhydrated or dehydrated to 75–80% of the initial body mass by withholding water for 3–4 days during exposure to intense solar radiation. The daily blood temperature means and maxima of were significantly higher in dehydration than in euhydration, but 40°C was rarely exceeded even during the hot hours of the day. Selective brain cooling occurred in euhydration, but its extent was small when blood temperature was below 39.5°C. In dehydration, however, selective brain cooling was frequent and the standard response when blood temperature exceeded 39°C. We believe that selective brain cooling contributes to the inhibition of evaporative heat loss, which is the primary cause of the higher blood temperature in dehydration. Rapid rehydration with cold water induced long-lasting depression of blood temperature. No evidence was found for mechanisms attenuating the subsequent decrease of brain temperature which occurred a few minutes after the uptake of cold water. Received: 9 March 1998 / Received after revision: 22 May 1998 / Accepted: 30 May 1998     

Unloading does not increase brown-adipose-tissue activity in rat pups

Our previous study has shown that chronic hindlimb suspension leads to an increase in both the thermogenic capacity and the activity in brown adipose tissue (BAT) of adult rats (Yamashita, H., Ohira, Y., Wakatsuki, T., Yamamoto, M., Kizaki, T., Oh-ishi, S., Sato, Y. and Ohno, H. (1995). J. Appl. Physiol. 78: 384-387). In order to examine if unloading also increases the BAT activity in rat pups, the hindlimbs in the suspended pups were unloaded by tail suspension beginning on postnatal day 4 and suspended until day 21. The thermogenic activity (which was assessed by guanosine 5'-diphosphate binding to BAT mitochondria) was markedly lower in 21-day-old suspended pups than in 21-day-old control pups, although there was no difference in uncoupling protein 1 (UCP1) content or UCP1 mRNA expression in the BAT mitochondrial fraction between both pups. Likewise, there was no disparity in either adrenal or thymus mass between the control and suspended pups throughout the experiment. These results suggest that, in contrast to adult rats, chronic hindlimb suspension leads to a decrease in the thermogenic activity in BAT of rat pups possibly for reason that pups are less susceptible to the stress of unloading.     

Temperature- and capsaicin-sensitive nerve fibers in brown adipose tissue attenuate thermogenesis in the rat

 We examined the function of putative sensory fibers that are contained in intercostal nerves and innervate interscapular brown adipose tissue (IBAT) in urethane-anesthetized rats. Warming the IBAT to 40–44°C with two small heaters placed bilaterally on the skin above it attenuated the subsequent noradrenaline-induced thermogenesis (NIT) of the IBAT. In this range of warming, higher IBAT temperatures resulted in more attenuation. Denervation of IBAT blocked the effect of thermal stimulation on the NIT. Thus, activation of nerve fibers in IBAT that are sensitive to warmth or to the nociceptive effects of heat probably attenuated the NIT. Since the putative sensory fibers in the IBAT contain calcitonin gene-related peptide (CGRP) and substance P, which are thought to act in peripheral tissues, we tested the effects of injection of these neuropeptides into the IBAT. Administration of 5.2 nmol CGRP but not substance P or vehicle saline mimicked the effect of thermal stimulation of IBAT. As the neuropeptide-containing primary sensory neurons are characterized by their sensitivity to capsaicin, we also tested its effects (1 mg/kg, s.c.) and found that it also attenuated the NIT. Denervation of the IBAT or pretreatment with capsazepine, a capsaicin receptor antagonist, blocked the effect of capsaicin. We propose that temperature- and capsaicin-sensitive nerve fibers release CGRP to attenuate the NIT of brown adipocytes. Received: 14 May 1998 / Received after revision: 6 August 1998 / Accepted: 11 August 1998     

Effects of chronic administration of noradrenaline and glucagon on in vitro brown adipose tissue thermogenesis.

The in vitro brown adipose tissue (BAT) oxygen consumption stimulated by noradrenaline (NA) or glucagon (G) was significantly lower in chronically NA-treated rats and that of G-treated rats did not differ as compared with that of vehicle-treated control animals. In vitro thermogenic response of BAT in NA-treated rats was consistent with that induced by cold acclimation.     

Cell surface measurements of ATP release from single pancreatic β cells using a novel biosensor technique

 To examine the possibility that ATP modulates insulin secretion by an autocrine mechanism, we measured the local concentration of released ATP at the surface of a single pancreatic β cell by a new biosensor technique, using PC12 cells expressing ligand-gated cation channels, P2X₂ receptors. Upon application of glucose or glibenclamide, a series of current spikes, whose amplitude equates to an ATP concentration of over 25 μM, were recorded from a PC12 cell using the whole-cell patch-clamp technique, when placed near a rat pancreatic β cell at 37°C. The current response was inhibited by cooling (below 30°C) or by applying an ATP-hydrolysing enzyme (apyrase) or a P2 receptor blocker (suramin). Thus, it is concluded that pancreatic β cells secrete ATP in response to glucose stimulation, thereby increasing the ATP concentration close to the cell surface sufficiently high enough to enhance insulin secretion from the pancreatic β cells. Received: 8 May 1998 / Received after revision: 4 August 1998 / Accepted: 10 August 1998     

Adenovirus-mediated gene expression in isolated rat pancreatic acini and individual pancreatic acinar cells

 In this study we have examined the feasibility of using replication-deficient recombinant adenoviral vectors to transfer and express genes in pancreatic acinar cellsin vitro. We infected primary cultures of both isolated pancreatic acini and individual acinar cells with a recombinant adenovirus containing the coding sequence for β-galactosidase. Our data demonstrate that recombinant adenoviruses readily infect pancreatic acinar cellsin vitro. Close to 100% infection and maximal β-galactosidase expression were obtained, when acini or acinar cells were infected with 5×10⁶ or 10⁶ plaque-forming units (pfu) of virus per millitre of acini or acinar cell suspension, respectively. Examination of the time-course of β-galactosidase expression showed that there was a lag of approximately 6 h before β-galactosidase levels increased. Thereafter β-galactosidase expression increased rapidly. By 20 h post-infection β-galactosidase activity had increased from undetectable levels to 2.5–3.0 units/mg of cellular protein. Acini/acinar cells maintained a robust secretory response after adenoviral infection. The cholecystokinin-octapeptide (CCK8) dose/response curves for amylase secretion for acini and acinar cells infected with 5×10⁵ and 1×10⁵ pfu/ml of virus, respectively, were biphasic, with maximal amylase secretion being stimulated by 1 nM CCK8. In addition, the dose/response curves were identical to those obtained from control, sham-infected, acini/acinar cells. Our findings indicate that replication-deficient recombinant adenoviral vectors will be excellent tools to transfer and express genes in isolated pancreatic acini or acinar cells. Received: 27 March 1998 / Received after remission: 6 May 1998 / Accepted: 7 May 1998     

Psychophysical study of noxious and innocuous cold discrimination in monkey

 Psychophysical evidence shows that humans are better able to distinguish differences in the intensity of cutaneous temperature in the cool range than in the noxious cold range. In order to compare these human perceptual findings with physiological data from non-human primates, we performed similar psychophysical experiments of cold perception in monkeys. Two adult male rhesus monkeys were trained to detect cooling shifts from baseline temperatures between 0° and 22°C applied to the face with a 1-cm² contact thermode. Detection thresholds were determined using the method of constant stimuli for one monkey and an adaptive psychophysical algorithm which insured constant behavioral performance for the other monkey. Results showed that both monkeys detected significantly smaller temperature decreases from innocuous cool baselines (i.e., 22° and 16°C) than from noxious and near-noxious baselines (10°, 6°, 0°C). Similarly, the latencies for detecting the cooling shifts were shorter and less variable in the innocuous cool range than in the noxious cold range. The observation of more precise discrimination of innocuous cool than noxious cold temperatures in monkeys is consistent with human psychophysical data. Thus, these data suggest that differential patterns of neuronal activity evoked by cool and noxious cold cutaneous stimuli, observed in peripheral afferents as well as in the central nervous system of monkey and cat, probably also exist in the human. Received: 29 June 1998 / Accepted: 19 October 1998     

Effect of Postnatal Age and a β3-Adrenergic Agonist (Zeneca D7114) Administration on Uncoupling Protein-1 Abundance in the Lamb

We examined the effect of time after birth and beta(3)-adrenergic agonist (Zeneca D7114) administration on uncoupling protein-1 (UCP1) abundance and thermoregulation in the lamb. Forty twin lambs, all born normally at term, were maintained at a cold ambient temperature of between 3 and 8 degrees C. At 0.5, 1.75, 5.25, 11.25 and 23.25 h after birth eight sets of twins were fed 20 ml of formula milk +/- 10 mg kg(-1) of beta(3)-adrenergic agonist, and 45 min after feeding brown adipose tissue (BAT) was sampled. Colonic temperature was measured and BAT analysed for UCP1 abundance, GDP-binding to mitochondrial protein (i.e. thermogenic activity) and catecholamine content. Colonic temperature declined between 1.25 and 6 h from 40.2 degrees C to 39.2 degrees C and then increased to 39.8 degrees C at 12 h, but increased after feeding at all ages. UCP1 abundance increased from 1.25 h after birth, to peak at 2 h after birth in controls, compared with 6 h after birth in beta(3)-adrenergic agonist-treated lambs. The level of GDP-binding to mitochondrial protein did not change significantly with age but was increased by beta(3)-adrenergic agonist treatment. The noradrenaline (norepinephrine) content of BAT increased between 1.25 and 12 h after birth, irrespective of beta(3)-adrenergic agonist administration. The total weight of perirenal BAT plus its lipid, protein and mitochondrial protein content declined over the first 6 h of life. UCP1 development continues over the first 24 h of neonatal life, and can be manipulated by beta(3)-adrenergic agonist administration. This may represent one method of improving thermoregulation in newborn lambs. Experimental Physiology (2001) 86.1, 65-70.     

Short photoperiod enhances thermogenic capacity in Brandt's voles

Environmental cues play important roles in the regulation of an animal's physiology and behavior. In the present study, we examined the effects of short photoperiod (SD) on body weight as well as on several physiological, hormonal, and biochemical measures indicative of thermogenic capacity to test our hypothesis that short photoperiod stimulates increases in thermogenesis without cold stress in Brandt's voles. SD voles showed increases in basal metabolic rate (BMR) and nonshivering thermogenesis (NST) during the 4-week photoperiod acclimation. At the end, these voles (SD) had lower body weights, higher levels of cytochrome C oxidase (COX) activity and mitochondrial uncoupling protein-1 (UCP1) contents in brown adipose tissues (BAT), and higher concentrations of serum tri-iodothyronine (T3) and thyroxine (T4) compared to LD voles. No differences were found between male and female voles in any of the above-mentioned measurements. Together, these data indicate that SD experience enhances thermogenic capacity similarly in males and females of Brandt's voles.     

Cold-induced thermogenesis in hypothyroid rats

Hypothyroidism was induced in adult rats by oral administration of methimazole. Euthyroid and hypothyroid rats were maintained at 23 °C or exposed at 6 °C for 3 weeks. Both euthyroid and hypothyroid rats maintained at 23 °C had similar interscapular brown adipose tissue (BAT) composition and thermogenic activity. Cold-exposed hypothyroid rats showed the same interscapular BAT mass and gross tissue composition as cold-exposed euthyroid animals, but the interscapular BAT of cold-exposed hypothyroid rats did not show the characteristic increase in GDP binding, and the increase in mitochondrial mass was lower than in euthyroid rats. From these results we conclude that thyroid hormones do not influence BAT significantly when thermogenic requirements are moderate, but they participate in the trophic response of the tissue when thermogenic requirements are intense. This thyroid hormone participation in the BAT trophic response occurs at the mitochondrial level, both in quantitative (mitochondrial mass) and qualitative (GDP-binding) aspects.     

Rapid effects of cytochalasin-D on contraction and intracellular calcium in single rat ventricular myocytes

 Contraction and intracellular calcium ([Ca²⁺]_i) transients were recorded using a video edge detector and fluorescence spectrophotometry, respectively, in rat ventricular myocytes at 22–24°C stimulated at a frequency of 1 Hz. Application of the F-actin disrupter cytochalasin-D (Cyt-D) caused a large reduction in the amplitude of contraction and a small increase in the [Ca²⁺]_i transient. These responses began within a few seconds of application and were complete after 2 min of exposure. Phase-plane relationships of contraction and [Ca²⁺]_i were consistent with cytochalasin-D causing a decrease in myofilament responsiveness to Ca²⁺. Received: 18 May 1998 / Received after revision: 8 July 1998 / Accepted: 13 July 1998     

Pituitary and autonomic responses to cold exposures in man

This review presents hormonal responses to various cold exposures and their calorigenic effects in man and some animals. Previous studies in rats have shown that cold exposures activate the hypothalamic-pituitary-thyroid axis. Increased thyroid hormone concentrations lead to heat production via general stimulation of metabolism (obligatory thermogenesis) and possibly via activation of thyroid hormone receptors and uncoupling protein 1 (UCP 1) and deiodinase enzyme genes in the brown adipose tissue (BAT). In human subjects long-term cold exposures do not seem to activate the pituitary-thyroid axis, but rather accelerate the elimination of triiodothyronine (T3), leading to low serum concentrations of free T3 hormone. In corollary to this a hypothyreotic condition with increased serum thyroid-stimulating hormone and impaired mood and cognitive performance can be observed after long-term cold exposures such as wintering. During cold exposures the sympathetic nerve system is activated and noradrenaline is released to blood circulation and to BAT, where it leads to production of cAMP, lipolysis and free fatty acids. Free fatty acids open the mitochondrial proton channel protein in BAT. Protons enter the mitochondria and inhibit ATP synthesis (uncoupling). By this way energy is transformed into heat (facultatory or adaptive thermogenesis). In adult human subjects the amount of BAT is small and adaptive thermogenesis (non-shivering thermogenesis) has a smaller role. UCP 1 with other uncoupling proteins may have other functions in the control of body weight, sugar balance and formation of reactive oxygen species.     

Brown adipose tissue of rats with obesity-inducing ventromedial hypothalamic lesions

Male and female Holtzman rats were made hyperphagic and obese with bilateral radiofrequency heat lesions of the ventromedial hypothalamic (VMH) area. When VMH rats were maintained at 28 degrees C, their brown adipose tissue (BAT) DNA, protein, and cytochrome oxidase contents were normal although more stored lipid was present, as judged from a threefold increase in wet weight. Thermogenic activity of BAT mitochondria was normal in male VMH rats, as judged from the unchanged level of guanosine diphosphate (GDP) binding (known to be a sensitive index of the functional activity of the thermogenic proton conductance pathway), and reduced in female VMH rats. When rats with VMH lesions were exposed to cold (4 degrees C for 24 h), the visible hyperemia of their BAT and normal large increase in mitochondrial GDP binding indicated normal thermogenic responsiveness. We conclude that the medial nuclei of the hypothalamus and associated afferent or efferent nerve tracts do not represent an essential central nervous system link for cold-induced, sympathetic-mediated activation of BAT thermogenesis. It is possible, however, that diet-induced, sympathetic-mediated activation of BAT function and growth might require an intact VMH region because no enhancement of BAT mitochondrial function normally associated with hyperphagia was detected in these hyperphagic VMH-lesioned animals.