Low intermale aggression associated with small submandibular and preputial glands in goldthioglucose-obese mice.

Goldthioglucose (GTG)-induced obese mice exhibited lower intermale aggression against an unfamiliar mouse, and had smaller submandibular and preputial glands than lean controls. The GTG-treated mice, which were pair-grown with control mice, also showed the same abnormalities. Therefore, obesity itself was not the cause of these abnormalities. One-h cohabitation with an unfamiliar mouse reduced the number of granules in the granular convoluted tubules of the submandibular glands of control mice. This histological change was not observed in the GTG-obese glands. These results suggest that the absence of typical intraspecific social behavior in male GTG-obese mice was associated with the failure to release a factor or factors from the submandibular glands.     

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.     

Inhibition by hypophysectomy of the hyperphagia and obesity following gold thioglucose

The effect of hypophysectomy in mice previously treated with gold thioglucose (GTG) was studied with respect to changes in food intake and development of obesity. As expected, all mice treated with GTG alone exhibited lesions in the ventromedial hypothalamus (VMH), hyperphagia and obesity. Hypophysectomy of GTG treated mice prevented the appearance of hyperphagia and obesity. Daily administration of the adrenal corticoid, cortisone, completely restored the hyperphagia and obesity in GTG treated hypophysectomized mice. The amounts of cortisone used did not appreciably affect food intake or body weights of normal, hypophysectomized or GTG treated mice. These findings indicate that the hypothalamic hyperphagia and obesity which normally follows the administration of GTG is dependent on a functional pituitary gland. Furthermore, the specific ability of an adrenal corticoid to completely restore the hyperphagia and obesity of GTG treated hypophysectomized mice in the absence of other pituitary factors, suggests that the pituitary adrenal axis serves as an important link in the regulatory mechanism for control of feeding behavior in the mouse.     

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.     

Norepinephrine turnover in obese (ob/ob) mice: effects of age, fasting, and acute cold

The possibility that low sympathetic nervous system (SNS) activity in brown adipose tissue (BAT) of 8-wk-old obese (ob/ob) mice results from their gross obesity at that age was investigated. Norepinephrine (NE) turnover, an estimator of SNS activity, was measured in BAT and other organs of 2-wk-old preobese ob/ob mice, and at 4 and 8 wk of age. Rates of NE turnover were 36% slower in BAT of preobese ob/ob mice than in lean littermates and remained slow in their BAT at 4 (-66%) and 8 (-56%) wk of age. In heart, rates of NE turnover were 48% slower in preobese ob/ob mice than in lean littermates, but the difference diminished at 4 (-21%) and 8 (-16%) wk of age. Rates of NE turnover in white adipose tissue, liver, and pancreas of obese mice were generally comparable with rates in these organs of lean mice. Effects of fasting (24 h) and acute cold exposure (14 degrees C for 8 h) were also examined. In general, fasting lowered and cold exposure elevated NE turnover equally in obese and lean mice. Ob/ob mice housed at 23-25 degrees C exhibit low SNS activity in their BAT prior to the onset of gross obesity, even though SNS activity in their BAT responds normally to an acute cold stress. This low SNS activity probably contributes to their subsequent high efficiency of energy retention.     

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.     

Cytochrome oxidase activity in brown fat varies with reproductive response and use of torpor in deer mice

Reproductive responses and thermogenic properties of brown adipose tissue (BAT) were evaluated in individuals from an outbred population of deer mice (Peromyscus maniculatus nebrascensis) after 10 weeks exposure to short photoperiod (8:16 light:dark) and cold ambient temperature (2 degrees C). Deer mice populations are composed of phenotypes that differ in their reproductive response to environmental cues. These phenotypes also differ in body temperature regulation as indicated by their use of daily torpor. By comparing BAT responses among individuals of different phenotypes, we were able to assess the association between environmentally induced changes in reproduction and metabolism. Short/cold days caused increased proliferation of BAT and higher cytochrome oxidase activity. However, the magnitude of these changes varied with reproductive phenotype and use of daily torpor. BAT weight in short/cold day exposed males with normal sized testes more than doubled while total cytochrome oxidase activity increased by 30% as compared to controls. In contrast, short/cold day exposed deer mice with atrophic testes that employed daily torpor exhibited a 64% increase in BAT weight and 100% increase in total cytochrome oxidase activity, compared to control mice. Cytochrome oxidase activity in nontorpid deer mice with atrophic testes was intermediate to these two groups. Our results demonstrate a response of BAT to short/cold days that varies with individual reproductive response. This finding suggests that there exists a common integrative mechanism for temperature and photoperiod to regulate both seasonal reproductive and metabolic adjustments.     

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.     

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.     

Nitric oxide and thermogenic function of brown adipose tissue in rats

To clarify the effects of cold acclimation and immobilization stress adaptation of rats on nitric oxide (NO) activity in interscapular brown adipose tissue (BAT), we incubated neatly diced (1-mm(3) blocks) BAT in a metabolic chamber for respiration, measured oxygen consumption using a Clark electrode, and estimated NO release in the buffer medium by measuring nitrite plus nitrate (NO(x)) using the Griess method (diazotization reaction). The production of NO(x) in the buffer medium confirmed that BAT releases NO, as there is no other source of NO(x) in the system. The NO activity was observed in the basal condition and increased with noradrenaline stimulation, showing a correlation with oxygen consumption in the warm (25 degrees C)-acclimated control rats. Cold acclimation (5 degrees C, 5 weeks) or immobilization stress adaptation (3 h daily, 25 degrees C, 5 weeks) caused enhanced NO activity in the basal condition in comparison with the control. We suggest that NO is involved in enhancement of the thermogenic functions of BAT in rats.     

Differential Control of the Sympathetic Nervous System by Leptin: Implications for Obesity

1. Leptin is a hormone that is secreted by adipocytes and delivered to the brain to regulate appetite and energy expenditure. Other effects of leptin include activation of the sympathetic nervous system and an increase in arterial pressure.2. Mounting evidence suggests that the sympathetic nervous system subserving different tissues is differentially controlled by leptin. For instance, leptin-induced regional increases in sympathetic nerve activity do not respond uniformly to baroreflex activation and hypothermia.3. In several mouse models of obesity, the ability of leptin to increase renal sympathetic nerve activity is preserved, despite resistance to leptin's effect on food intake, body weight and thermogenic sympathetic tone. Furthermore, obese mice also retain the increase in arterial pressure in response to leptin.3. Although they display a lack of metabolic responses to leptin, animal models of obesity preserve renal sympathetic and arterial pressure responses that potentially cause the adverse cardiovascular consequences of obesity. Thus, it is possible that excess leptin contributes to cardiovascular complications, even when a subject shows metabolic resistance to leptin.     

Sex-associated differences in cold-induced UCP1 synthesis in rodent brown adipose tissue

 The effects of acute and chronic acclimation to cold on uncoupling protein 1 (UCP1) levels, as well as on GDP-binding to mitochondria, cytochrome c oxidase activity and mitochondrial protein concentration in brown adipose tissue (BAT) of intact male and female rats have been analyzed. Results reveal that females rats are more sensitive to cold because their threshold temperature for the thermogenic response is set at a higher value (around 22°C) than that of males (around 18°C), hence leading to differences in BAT UCP1 levels between the sexes at different environmental temperatures. In vitro experiments showed that steroid hormones, β-estradiol, estrone and progesterone, can reduce norepinephrine-induced UCP1 synthesis in brown adipocytes differentiated in primary culture. Thus the different sex-associated response of cold-induced thermogenesis in rats does not appear to be explained by a direct action of sex steroids upon the adipocyte, implying that other factors in the thermogenic regulatory system must be involved. Received: 23 March 1998 / Received after revision: 20 May 1998 / Accepted: 21 May 1998     

Involvement of thermosensitive TRP channels in energy metabolism

To date, 11 thermosensitive transient receptor potential (thermo-TRP) channels have been identified. Recent studies have characterized the mechanism of thermosensing by thermo-TRPs and the physiological role of thermo-TRPs in energy metabolism. In this review, we highlight the role of various thermo-TRPs in energy metabolism and hormone secretion. In the pancreas, TRPM2 and other TRPs regulate insulin secretion. TRPV2 expressed in brown adipocytes contributes to differentiation and/or thermogenesis. Sensory nerves that express TRPV1 promote increased energy expenditure by activating sympathetic nerves and adrenaline secretion. Here, we first show that capsaicin-induced adrenaline secretion is completely impaired in TRPV1 knockout mice. The thermogenic effects of TRPV1 agonists are attributable to brown adipose tissue (BAT) activation in mice and humans. Moreover, TRPA1- and TRPM8-expressing sensory nerves also contribute to potentiation of BAT thermogenesis and energy expenditure in mice. Together, thermo-TRPs are promising targets for combating obesity and metabolic disorders.     

Effect of acute cold-exposure on norepinephrine turnover and thermogenesis in brown adipose tissue and metabolic rate in MSG-induced obese mice

Mice treated neonatally with monosodium-L-glutamate (MSG) are known to develop into obese adults without hyperphagia, which are characterized by the reduced levels in the resting metabolic rate (RMR) and the thermogenesis of brown adipose tissue (BAT) in the thermoneutral environment. The present study revealed that an acute cold-exposure (5 degrees C, 1 h) of these animals resulted in the increase in norepinephrine turnover and mitochondrial-5'-diphosphate (GDP) binding in the interscapular BAT as well as the guanosine RMR, suggesting a normal thermogenic responsiveness of BAT to cold.     

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.     

Effects of cold acclimation on cold-induced changes in lipid metabolism of rat brown adipose tissue.

Effects of acute cold exposure (0 degree C, 12 h) on lipid metabolism of the interscapular brown adipose tissue (BAT) were studied in warm controls (25 degrees C) (WC) and cold-acclimated rats (5 degrees C, 4 weeks) (CA). 1) We confirmed that cold acclimation brought about decreased BAT triglyceride (TG) level and increased BAT phospholipid (PL) level. The indexes of unsaturation, such as unsaturation index and polyunsaturated fatty acids/saturated fatty acids ratio, of bulky fatty acids (FA) (palmitic, stearic, oleic, and linoleic acids) decreased in TG, while those increased in PL. Rare FA (eicosadienoic acid, homo-gamma-linoleinic acid, lignoceric acid) level, which were about five times higher in PL than TG in WC, decreased in both TG and PL in CA. 2) Effect of acute cold exposure in WC: The level as well as the amount of TG in BAT was greatly reduced and the indexes of unsaturation in TG-FA increased. The most part of reduced TG (85%) was explained by the bulky FA. Rare FA explained only 1.1% of reduced TG. PL level in BAT did not change, but its amount decreased. The indexes of unsaturation increased in PL-FA. 3) Effect of acute cold exposure in CA: CA was placed in warm temperature of 25 degrees C before cold exposure. In contrast with the cold-reduced TG in WC, either the level or the total amount of TG in BAT did not change, but the indexes of unsaturation in TG-FA decreased. The level as well as the total amount of PL in BAT increased. The arachidonic index and arachidonate in PL-FA decreased, but the indexes of unsaturation did not change. These results indicate as a whole that acute cold exposure as well as cold acclimation influences BAT lipid metabolism in FA compositions as well as amount of TG and PL, suggesting that such changes are related to thermogenic activity of this tissue.     

Metabolic activity of brown adipose tissue in T3-treated hamsters.

Triiodothyronine (T3) treatment induced marked hypertrophy of the brown adipose tissue (BAT), similar to that observed in cold-acclimated animals, although partly due to fat deposition. Similar to cold acclimation, T3 treatment also increased the oxidation of succinate by tissue slices without concomitant increase in isolated mitochondria. It is therefore suggested that thyroid hormones, like cold acclimation, effect conformational changes in the mitochondria, leading to greater expression of the succinic oxidase in the tissue. T3 treatment was not followed by any change in the respiratory activity of tissue slices in the presence of alpha-GP. Likewise, no change was found either in other oxidative activities tested in isolated mitochondria (e.g., NADH and cytochrome oxidases) or in the concentration of the components of the electron transport chain in the mitochondria.     

Reduced norepinephrine turnover in brown adipose tissue of ob/ob mice

Obese (ob/ob) mice have a lower thermogenic capacity than lean mice. The possible role of brown adipose tissue (BAT) in this defect was investigated. Lean and obese mice were exposed to 33 (thermoneutral), 25, or 14 degrees C for up to 3 wk. BAT cytochrome oxidase activity and numbers of Na+-K+-ATPase enzyme units, enzymes involved in thermogenesis, were similar at 33 or 25 degrees C. Chronic exposure to 14 degrees C increased these enzymes 34 and 62%, respectively, in lean mice and nearly 150% in obese mice. Sympathetic nervous system activity, which stimulates thermogenesis in BAT, was evaluated by measuring norepinephrine (NE) turnover. At 25 degrees C, NE turnover rate in BAT of obese mice was only 40% as rapid as in BAT of lean mice. Chronic exposure to 33 degrees C depressed NE turnover in BAT of lean mice, but not in obese mice, whereas exposure to 14 degrees C accelerated NE turnover in both lean and obese mice. Lower sympathetic nervous system activity in BAT of obese mice at 25 degrees C is likely a major factor in their reduced nonshivering thermogenesis and resultant high efficiency of energy storage.     

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.