RESUMO
Previous studies have shown that very low dose, acute, single peripheral leptin injections fully activate arcuate nucleus signal transducer and activator of transcription 3 (STAT3), but ventromedial hypothalamus (VMH) pSTAT3 continues to increase with higher doses of leptin that inhibit food intake. The lowest dose that inhibited intake increased circulating leptin 300-fold whereas food intake is inhibited by chronic peripheral leptin infusions that only double circulating leptin. This study examined whether the pattern of hypothalamic pSTAT3 was the same in leptin-infused rats as in leptin-injected rats. Male Sprague-Dawley rats received intraperitoneal infusions of 0, 5, 10, 20, or 40 µg leptin/day for 9 days. The highest dose of leptin increased serum leptin by 50-100%, inhibited food intake for 5 days, but inhibited weight gain and retroperitoneal fat mass for 9 days. Energy expenditure, respiratory exchange ratio, and brown fat temperature did not change. pSTAT3 was quantified in hypothalamic nuclei and the nucleus of the solitary tract (NTS) when food intake was inhibited and when it had returned to control levels. There was no effect of leptin on pSTAT3 in the medial or lateral arcuate nucleus or in the dorsomedial nucleus of the hypothalamus. VMH pSTAT3 was increased only at day 4 when food intake was inhibited, but NTS pSTAT3 was increased at both 4 and 9 days of infusion. These results suggest that activation of leptin VMH receptors contributes to the suppression of food intake, but that hindbrain receptors contribute to a sustained change in metabolism that maintains a reduced weight and fat mass.NEW & NOTEWORTHY Low-dose, chronic peripheral infusions of leptin produced an initial, transient inhibition of food intake that correlated with signal transducer and activator of transcription 3 (STAT3) activation in the ventromedial hypothalamus (VMH) and nucleus of the solitary tract (NTS). When intake normalized, but weight remained suppressed, the NTS was the only area that remained activated. These data suggest that leptin's primary function is to reduce body fat, that hypophagia is a means of achieving this and that different areas of the brain are responsible for the progressive response.
Assuntos
Leptina , Fator de Transcrição STAT3 , Ratos , Masculino , Animais , Leptina/metabolismo , Ratos Sprague-Dawley , Fator de Transcrição STAT3/metabolismo , Hipotálamo/metabolismo , Núcleo Solitário/metabolismo , Tecido Adiposo Marrom/metabolismo , Infusões Intraventriculares , Ingestão de Alimentos , Receptores para Leptina/metabolismoRESUMO
Rats offered free access to sucrose solution in addition to a sucrose-free composite diet develop leptin resistance whereas those consuming a similar amount of sucrose from a dry diet remain leptin responsive. Here we tested whether rats consuming a complete high sucrose diet in liquid form also became leptin resistant. Female Sprague Dawley rats were offered a sucrose free diet (NS), a dry high sucrose diet (HS), NS diet plus 30% sucrose solution (LiqS), NS diet in liquid form (NSLiq) or HS diet in Liquid form (HSLiq). After 30 days LiqS rats were leptin resistant, but all other groups were leptin responsive even though HSLiq rats consumed as much sucrose as LiqS rats and NSLiq rats had the greatest amount of body fat. Therefore, development of leptin resistance is dependent upon the consumption of sucrose independent of any other nutrients. Because LiqS rats consume sucrose throughout the day and night we tested whether limiting sucrose solution access to either the light or dark period prevented development of leptin resistance. Leptin resistant LiqS rats were either given free access to sucrose, had access to sucrose only at night or had access only during the day. The intake of rats with limited access was supplemented to the level of those with free access by tube-feeding. The results of this study show that leptin resistance of LiqS rats is independent of when the sucrose is consumed and is unrelated to total energy intake, body fat mass or serum leptin concentration.
Assuntos
Sacarose Alimentar , Leptina , Feminino , Ratos , Animais , Ratos Sprague-Dawley , Ingestão de Energia , Dieta , Peso CorporalRESUMO
Others have shown that leptin and cholecystokinin (CCK) act synergistically to suppress food intake. Experiments described here tested whether leptin in the ventromedial hypothalamus (VMH) contributes to the synergy with peripheral CCK in male Sprague Dawley rats. A subthreshold injection of 50-ng leptin into the VMH 1 h before a peripheral injection of 1 µg/kg CCK did not change the response to CCK in rats offered chow or low-fat purified diet, but did exaggerate the reduction in intake of high-fat diet 30 min and 1 h after injection in rats that had been food deprived for 8 h. By contrast, deletion of leptin receptor-expressing cells in the VMH using leptin-conjugated saporin (Lep-Sap) abolished the response to peripheral CCK in chow-fed rats. Lateral ventricle injection of 2-µg leptin combined with peripheral CCK exaggerated the inhibition of chow intake for up to 6 h in control rats treated with Blank-saporin, but not in Lep-Sap rats. Blank-Saporin rats offered low- or high-fat purified diet also demonstrated a dose-response inhibition of intake that reached significance with 1 µg/kg of CCK for both diets. CCK did not inhibit intake of Lep-Sap rats in either low- or high-fat-fed rats. Thus, although basal activation of VMH leptin receptors makes a significant contribution to the synergy with CCK, increased leptin activity in the VMH does not exaggerate the response to CCK in intact rats offered low-fat diets, but does enhance the response in those offered high-fat diet.NEW & NOTEWORTHY Leptin is a feedback signal in the control of energy balance, whereas cholecystokinin (CCK) is a short-term satiety signal that inhibits meal size. The two hormones synergize to promote satiety. We tested whether leptin receptors in the ventromedial nucleus of the hypothalamus (VMH) contribute to the synergy. The results suggest that there is a requirement for a baseline level of activation of leptin receptors in the VMH in order for CCK to promote satiety.
Assuntos
Leptina , Receptores para Leptina , Animais , Colecistocinina/farmacologia , Leptina/metabolismo , Leptina/farmacologia , Masculino , Ratos , Ratos Sprague-Dawley , Receptores para Leptina/genética , Receptores para Leptina/metabolismo , Saporinas , Núcleo Hipotalâmico Ventromedial/metabolismoRESUMO
This experiment investigated which hypothalamic nuclei were activated by a dose of leptin that inhibited food intake. Foodnot intake, energy expenditure, respiratory exchange ratio (RER), and intrascapular brown adipose tissue (IBAT) temperature were measured in male and female Sprague Dawley rats for 36 h following an intraperitoneal injection of 0, 50, 200, 500, or 1,000 µg leptin/kg with each rat tested with each dose of leptin in random order. In both males and females, RER and 12-h food intake were inhibited only by 1,000 µg leptin/kg, but there was no effect on energy expenditure or IBAT temperature. At the end of the experiment, phosphorylated signal transducer and activator of transcription 3 (pSTAT3) immunoreactivity was measured 1 h after injection of 0, 50, 500, or 1,000 µg leptin/kg. In male rats, the lowest dose of leptin produced a maximal activation of STAT3 in the Arc and nucleus of the solitary tract (NTS). There was no response in the dorsomedial hypothalamus, but there was a progressive increase in ventromedial nucleus of the hypothalamus (VMH) pSTAT3 with increasing doses of leptin. In female rats, there was no significant change in Arc and pSTAT3 NTS activation was maximal with 500 mg leptin/kg, but only the highest dose of leptin increased VMH pSTAT3. These results suggest that the VMH plays an important role in the energetic response to elevations of circulating leptin but do not exclude the possibility that multiple nuclei provide the appropriate integrated response to hyperleptinemia.NEW & NOTEWORTHY The results of this experiment show that doses of leptin too small to inhibit food intake produce a maximal response to leptin in the arcuate nucleus. By contrast the VMH shows a robust response that correlates with inhibition of food intake. This suggests that the VMH plays an important role in the energetic response to hyperleptinemia.
Assuntos
Ingestão de Alimentos/efeitos dos fármacos , Hipotálamo/metabolismo , Leptina/administração & dosagem , Fosforilação/efeitos dos fármacos , Fator de Transcrição STAT3/metabolismo , Tecido Adiposo Marrom/efeitos dos fármacos , Tecido Adiposo Marrom/metabolismo , Animais , Núcleo Arqueado do Hipotálamo/metabolismo , Temperatura Corporal/efeitos dos fármacos , Relação Dose-Resposta a Droga , Metabolismo Energético/efeitos dos fármacos , Feminino , Hipotálamo/química , Leptina/sangue , Masculino , Ratos , Ratos Sprague-Dawley , Fator de Transcrição STAT3/análise , Núcleo Solitário/metabolismo , Núcleo Hipotalâmico Ventromedial/metabolismoRESUMO
Rats consuming 30% sucrose solution and a sucrose-free diet (LiqS) become leptin resistant, whereas rats consuming sucrose from a formulated diet (HS) remain leptin responsive. This study tested whether leptin resistance in LiqS rats extended beyond a failure to inhibit food intake and examined leptin responsiveness in the hypothalamus and hindbrain of rats offered HS, LiqS, or a sucrose-free diet (NS). Female LiqS Sprague-Dawley rats initially only partially compensated for the calories consumed as sucrose, but energy intake matched that of HS and NS rats when they were transferred to calorimetry cages. There was no effect of diet on energy expenditure, intrascapular brown fat tissue (IBAT) temperature, or fat pad weight. A peripheral injection of 2 mg of leptin/kg on day 23 or day 26 inhibited energy intake of HS and NS but not LiqS rats. Inhibition occurred earlier in HS rats than in NS rats and was associated with a smaller meal size. Leptin had no effect on energy expenditure but caused a transient rise in IBAT temperature of HS rats. Leptin increased the phosphorylation of signal transducer and activator of transcription 3 (pSTAT3) in the hindbrain and ventromedial hypothalamus of all rats. There was a minimal effect of leptin in the arcuate nucleus, and only the dorsomedial hypothalamus showed a correlation between pSTAT3 and leptin responsiveness. These data suggest that the primary response to leptin is inhibition of food intake and the pattern of sucrose consumption, rather than calories consumed as sucrose, causes leptin resistance associated with site-specific differences in hypothalamic leptin signaling.
Assuntos
Hipotálamo/metabolismo , Leptina/metabolismo , Transdução de Sinais/efeitos dos fármacos , Sacarose/farmacologia , Animais , Feminino , Ratos , Ratos Sprague-Dawley , Transdução de Sinais/fisiologia , Sacarose/administração & dosagemRESUMO
Previous studies indicate that inhibition of food intake by leptin is mediated by an integrated response to activation of hypothalamic and hindbrain receptors. This study tested whether loss of hindbrain leptin receptor signaling changed sensitivity to forebrain leptin. Injections of leptin-conjugated saporin (Lep-Sap) into the medial nucleus of the solitary tract (NTS) were used to destroy hindbrain leptin receptor-expressing neurons of male Sprague-Dawley rats. Controls were injected with saporin conjugated with a nonsense peptide (Blk-Sap). Lep-Sap had no effect on daily food intake or body weight, but expression of phosphorylated signal transducer and activator of transcription 3 (pSTAT3) in the NTS following a peripheral injection of leptin was abolished 26 days after Lep-Sap injections. To test forebrain leptin sensitivity, Lep-Sap and Blk-Sap rats received third-ventricle injections of 0, 0.05, 0.1, 0.25, or 0.5 µg leptin. Food intake was inhibited by 0.25 and 0.5 µg leptin in Blk-Sap rats, but there was no significant effect of leptin on food intake of Lep-Sap rats. There was no difference in hypothalamic pSTAT3 in unstimulated conditions, but pSTAT3 was lower in the dorsomedial region of the ventromedial nucleus of the hypothalamus (VMH) of Lep-Sap rats compared with Blk-Sap rats following a third-ventricle injection of 0.25 µg leptin. These results are consistent with previous data showing that loss of VMH leptin receptor-expressing cells prevents weight loss caused by fourth-ventricle leptin infusion and show that the integrated response between the hindbrain and forebrain is heavily dependent on leptin activity in the VMH.
Assuntos
Ingestão de Alimentos/efeitos dos fármacos , Leptina/farmacologia , Neurônios/metabolismo , Prosencéfalo/efeitos dos fármacos , Receptores para Leptina/metabolismo , Rombencéfalo/metabolismo , Núcleo Hipotalâmico Ventromedial/metabolismo , Animais , Peso Corporal/efeitos dos fármacos , Masculino , Neurônios/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Receptores para Leptina/genéticaRESUMO
Leptin administration into the hindbrain, and specifically the nucleus of the solitary tract, increases phosphorylated signal transducer and activator of transcription 3 (pSTAT3), a marker of leptin receptor activation, in hypothalamic nuclei known to express leptin receptors. The ventromedial nucleus of the hypothalamus (VMH) shows the greatest response, with a threefold increase in pSTAT3. This experiment tested the importance of VMH leptin receptor-expressing neurons in mediating weight loss caused by fourth ventricle (4V) leptin infusion. Male Sprague-Dawley rats received bilateral VMH 75-nL injections of 260 ng/µL of leptin-conjugated saporin (Lep-Sap) or blank-saporin (Blk-Sap). After 23 days they were fitted with 4V infusion cannulas and 1 wk later adapted to housing in a calorimeter before they were infused with 0.9 µg leptin/day for 14 days. There was no effect of VMH Lep-Sap on weight gain or glucose clearance before leptin infusion. Leptin inhibited food intake and respiratory exchange ratio in Blk-Sap but not Lep-Sap rats. Leptin had no effect on energy expenditure or brown adipose tissue temperature of either group. Inguinal and epididymal fat were significantly reduced in leptin-treated Blk-Sap rats, but the response was greatly attenuated in Lep-Sap rats. VMH pSTAT3 was increased in leptin-treated Blk-Sap but not Lep-Sap rats. These results support the concept that leptin-induced weight loss results from an integrated response across different brain areas. They also support previous reports that VMH leptin receptors do not play a significant role in maintaining energy balance in basal conditions but limit weight gain during positive energy balance.
Assuntos
Quarto Ventrículo , Leptina/administração & dosagem , Leptina/farmacologia , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Receptores para Leptina/efeitos dos fármacos , Receptores para Leptina/metabolismo , Núcleo Hipotalâmico Ventromedial/metabolismo , Redução de Peso/efeitos dos fármacos , Tecido Adiposo/efeitos dos fármacos , Tecido Adiposo/crescimento & desenvolvimento , Animais , Temperatura Corporal/efeitos dos fármacos , Ingestão de Alimentos/efeitos dos fármacos , Glucose/metabolismo , Infusões Intraventriculares , Masculino , Ratos , Ratos Sprague-Dawley , Fator de Transcrição STAT3/biossíntese , Fator de Transcrição STAT3/genética , Saporinas/farmacologia , Núcleo Hipotalâmico Ventromedial/efeitos dos fármacosRESUMO
Previous studies suggest that weight loss occurs when leptin receptors in both the forebrain and hindbrain are activated. Experiments described here tested whether this integration is mediated through a neural connection or by leptin diffusion through the subarachanoid space. If the hypothalamus and hindbrain communicated through a neural pathway, then a very low dose of leptin infused directly into the nucleus of the solitary tract (NTS) would enhance the response to third ventricle (3V) leptin but would have no effect if infused into the fourth ventricle (4V). A 12-day infusion of 10 ng/24 h into the 4V or the NTS reduced body fat. Leptin at 5 ng/24 h into the 4V or NTS had no effect on food intake or body composition, but infusion of 5 ng of leptin/24 h into the NTS combined with a 3V injection of 0.1 µg of leptin inhibited food intake between 6 and 12 h after injection. Cumulative intake was inhibited for up to 36 h. 3V leptin had no effect on food intake of rats receiving the 4V leptin infusion. Similar results were found using infusions of 5 ng leptin/24 h and a 3V injection of 0.025 µg leptin. These data suggest that activation of leptin receptors in the NTS lowers the threshold for response to leptin in the forebrain through a neural network.
Assuntos
Tecido Adiposo/efeitos dos fármacos , Composição Corporal/efeitos dos fármacos , Ingestão de Alimentos/efeitos dos fármacos , Leptina/farmacologia , Núcleo Solitário/metabolismo , Animais , Metabolismo Energético , Quarto Ventrículo/metabolismo , Hipotálamo/metabolismo , Injeções Intraventriculares , Leptina/metabolismo , Masculino , Vias Neurais , Ratos , Ratos Sprague-Dawley , Receptores para Leptina/metabolismo , Terceiro Ventrículo/metabolismoRESUMO
Rats offered 30% sucrose solution plus chow or a sucrose-free diet develop leptin resistance within 4 weeks. This experiment tested whether leptin resistance was associated with the reward of sweet taste or the pre- or post-absorptive effects of consumption of simple carbohydrate. Male Sprague Dawley rats were offered a sucrose-free diet (NS), a diet containing 67% calories as sucrose (HS) or NS diet plus 30% sucrose (LS), 0.03% saccharin (Sacc) or 20% SolCarb® solution for 38 days. SolCarb® is a maltodextrin powder. Sacc rats initially drank more than LS rats, but intakes were the same after Day 20. SolCarb® and LS rats drank the same number of calories from Day 15 to the end of the experiment. SolCarb® and LS rats ate less dry food than other groups, but total energy intake was greater than that of NS, HS and Sacc groups and over 80% of their energy intake was from carbohydrate. Leptin responsiveness was tested on Days 27 and 32 with each rat acting as its own control. An i.p. injection of 2â¯mg/kg leptin inhibited food intake of NS, HS and Sacc rats, but had no effect on energy intake of LS or SolCarb® rats or on consumption of Sacc, sucrose or SolCarb® solution. At the end of the experiment all of the rats were insulin sensitive, had the same body composition and serum leptin concentrations. These data indicate that consumption of a calorie containing carbohydrate solution and not sweet taste drives the development of leptin resistance and suggest that there is lower threshold for inhibition of hunger than for inhibition of reward by leptin.
Assuntos
Carboidratos da Dieta/administração & dosagem , Sacarose Alimentar/efeitos adversos , Ingestão de Energia , Leptina/fisiologia , Animais , Dieta , Resistência à Insulina , Masculino , Ratos , Ratos Sprague-Dawley , Sacarina , PaladarRESUMO
Spontaneous intracerebral hemorrhage (ICH) produces the highest acute mortality and worst outcomes of all stroke subtypes. Hematoma volume is an independent determinant of ICH patient outcomes, making clot resolution a primary goal of clinical management. Herein, remote-limb ischemic post-conditioning (RIC), the repetitive inflation-deflation of a blood pressure cuff on a limb, accelerated hematoma resolution and improved neurological outcomes after ICH in mice. Parabiosis studies revealed RIC accelerated clot resolution via a humoral-mediated mechanism. Whereas RIC increased anti-inflammatory macrophage activation, myeloid cell depletion eliminated the beneficial effects of RIC after ICH. Myeloid-specific inactivation of the metabolic regulator, AMPKα1, attenuated RIC-induced anti-inflammatory macrophage polarization and delayed hematoma resolution, providing a molecular link between RIC and immune activation. Finally, chimera studies implicated myeloid CD36 expression in RIC-mediated neurological recovery after ICH. Thus, RIC, a clinically well-tolerated therapy, noninvasively modulates innate immune responses to improve ICH outcomes. Moreover, immunometabolic changes may provide pharmacodynamic blood biomarkers to clinically monitor the therapeutic efficacy of RIC.
Assuntos
Proteínas Quinases Ativadas por AMP/imunologia , Hematoma/imunologia , Pós-Condicionamento Isquêmico , Ativação de Macrófagos , Macrófagos/imunologia , Acidente Vascular Cerebral/imunologia , Proteínas Quinases Ativadas por AMP/genética , Animais , Hematoma/patologia , Hematoma/terapia , Macrófagos/patologia , Camundongos , Camundongos Knockout , Acidente Vascular Cerebral/patologia , Acidente Vascular Cerebral/terapiaRESUMO
Male rats offered 30% sucrose solution in addition to chow develop leptin resistance without an increase in energy intake or body fat. This study tested whether the leptin resistance was dependent on the physical form of the sucrose. Sprague-Dawley rats were offered a sucrose-free (NS) diet, a 66.6% of energy as sucrose (HS) diet, or the NS diet + 30% sucrose solution (LS). Sucrose intake of LS rats equaled that of HS rats, but total carbohydrate intake exceeded that of HS rats. After 33 days, male and female LS rats were resistant to the inhibitory effect of peripherally administered leptin on food intake. LS rats drank small, frequent meals of sucrose during light and dark periods, whereas HS rats consumed more meals during the dark than the light period and remained responsive to leptin. Diet did not affect daily energy intake or insulin sensitivity. There was a small increase in body fat in the female rats. Leptin sensitivity was restored within 5 days of withdrawal from sucrose in male LS rats. This rapid reversal suggested that leptin resistance was associated with the metabolic impact of drinking sucrose. An experiment was carried out to test whether activity of the hexosamine biosynthetic pathway and glycation of leptin signaling proteins were increased in LS rats, but the results were equivocal. A final experiment determined that female LS rats were leptin-resistant within 18 days of access to sucrose solution and that the small, but significant, increase in body fat was associated with increased adipocyte glucose utilization and insulin responsiveness, which may have been secondary to adipocyte leptin resistance.
Assuntos
Tecido Adiposo Branco/metabolismo , Sacarose Alimentar/toxicidade , Leptina/sangue , Adipócitos/metabolismo , Adiposidade , Administração Oral , Ração Animal , Fenômenos Fisiológicos da Nutrição Animal , Animais , Biomarcadores/sangue , Glicemia/metabolismo , Sacarose Alimentar/administração & dosagem , Sacarose Alimentar/sangue , Ingestão de Energia , Ácidos Graxos/sangue , Feminino , Insulina/sangue , Resistência à Insulina , Masculino , Estado Nutricional , Ratos Sprague-Dawley , Fatores Sexuais , Fatores de TempoRESUMO
This review summarizes the evidence derived from studies utilizing denervation procedures to demonstrate sympathetic control of white adipose tissue metabolism and body fat mass. A majority of the work demonstrating neural control of white fat was performed in the Bartness laboratory with Siberian hamsters as the predominant experimental model. These animals experience dramatic changes in body fat mass in response to changes in photoperiod, however, the mechanisms identified in hamsters have been reproduced or further elucidated by experiments with other animal models. Evidence for the role of sympathetic innervation contributing to the control of white adipocyte lipolysis and preadipocyte proliferation is summarized. In addition, evidence from denervation experiments for neural communication between different white fat depots as well as for a feedback control loop between sensory afferents from individual fat depots and sympathetic efferents to the same or distant white fat depots is discussed.
Assuntos
Tecido Adiposo Branco/fisiologia , Simpatectomia , Sistema Nervoso Simpático/fisiologia , Tecido Adiposo Branco/inervação , Animais , Cricetinae , PhodopusRESUMO
We previously reported that low-dose leptin infusions into the third or fourth ventricle that do not affect energy balance when given independently cause rapid weight loss when given simultaneously. Therefore, we tested whether hindbrain leptin enhances the response to forebrain leptin or whether forebrain leptin enhances the response to hindbrain leptin. Rats received fourth-ventricle infusions of saline or 0.01, 0.1, 0.3, or 0.6 µg leptin/day for 13 days. On days 9 and 13, 0.1 µg leptin was injected into the third ventricle. The injection inhibited food intake for 36 h in saline-infused rats but for 60 h in those infused with 0.6 µg leptin/day. Leptin injection increased intrascapular brown fat temperature in leptin-infused, but not saline-infused, rats. In a separate experiment, rats received third-ventricle infusions of saline or 0.005, 0.01, 0.05, or 0.1 µg leptin/day and fourth-ventricle injections of 1.0 µg leptin on days 9 and 13 Leptin injection inhibited food intake, respiratory exchange ratio, and 14-h food intake in rats infused with saline or the two lowest doses of leptin. There was no effect with higher-dose leptin infusions because food intake, body fat, and lean mass were already inhibited. These data suggest that activation of leptin receptors in the hindbrain enhances the response to third-ventricle leptin, whereas activation of forebrain leptin receptors does not enhance the response to fourth-ventricle leptin, consistent with our previous finding that weight loss in rats treated with fourth-ventricle leptin is associated with indirect activation of hypothalamic STAT3.
Assuntos
Quarto Ventrículo/efeitos dos fármacos , Leptina/administração & dosagem , Terceiro Ventrículo/efeitos dos fármacos , Terceiro Ventrículo/metabolismo , Animais , Peso Corporal/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ingestão de Alimentos/efeitos dos fármacos , Metabolismo Energético/efeitos dos fármacos , Quarto Ventrículo/metabolismo , Infusões Intraventriculares , Masculino , Ratos , Ratos Sprague-Dawley , Receptores para Leptina/metabolismoRESUMO
Rats and mice exposed to repeated stress or a single severe stress exhibit a sustained increase in energetic, endocrine, and behavioral response to subsequent novel mild stress. This study tested whether the hyper-responsiveness was due to a lowered threshold of response to corticotropin releasing factor (CRF) or an exaggerated response to a standard dose of CRF. Male Sprague-Dawley rats were subjected to 3h of restraint on each of 3 consecutive days (RRS) or were non-restrained controls. RRS caused a temporary hypophagia but a sustained reduction in body weight. Eight days after the end of restraint, rats received increasing third ventricle doses of CRF (0-3.0µg). The lowest dose of CRF (0.25µg) increased corticosterone release in RRS, but not control rats. Higher doses caused the same stimulation of corticosterone in the two groups of rats. Fifteen days after the end of restraint, rats were food deprived during the light period and received increasing third ventricle doses of CRF at the start of the dark period. The lowest dose of CRF inhibited food intake during the first hour following infusion in RRS, but not control rats. All other doses of CRF inhibited food intake to the same degree in both RRS and control rats. The lowered threshold of response to central CRF is consistent with the chronic hyper-responsiveness to CRF and mild stress in RRS rats during the post-restraint period.
Assuntos
Comportamento Animal/fisiologia , Hormônio Liberador da Corticotropina/farmacologia , Ingestão de Alimentos/efeitos dos fármacos , Restrição Física , Estresse Psicológico/fisiopatologia , Animais , Comportamento Animal/efeitos dos fármacos , Hormônio Liberador da Corticotropina/administração & dosagem , Masculino , Ratos , Ratos Sprague-Dawley , Terceiro Ventrículo/efeitos dos fármacosRESUMO
Previous studies have shown that very low-dose infusions of leptin into the third or the fourth ventricle alone have little effect on energy balance, but simultaneous low-dose infusions cause rapid weight loss and increased phosphorylation of STAT3 (p-STAT3) in hypothalamic sites that express leptin receptors. Other studies show that injecting high doses of leptin into the fourth ventricle inhibits food intake and weight gain. Therefore, we tested whether fourth-ventricle leptin infusions that cause weight loss are associated with increased leptin signaling in the hypothalamus. In a dose response study 14-day infusions of increasing doses of leptin showed significant hypophagia, weight loss, and increased hypothalamic p-STAT3 in rats receiving at least 0.9 µg leptin/day. In a second study 0.6 µg leptin/day transiently inhibited food intake and reduced carcass fat, but had no significant effect on energy expenditure. In a final study, we identified the localization of STAT3 activation in the hypothalamus of rats receiving 0, 0.3, or 1.2 µg leptin/day. The high dose of leptin, which caused weight loss in the first experiment, increased p-STAT3 in the ventromedial, dorsomedial, and arcuate nuclei of the hypothalamus. The low dose that increased brown fat UCP1 but did not affect body composition in the first experiment had little effect on hypothalamic p-STAT3. We propose that hindbrain leptin increases the precision of control of energy balance by lowering the threshold for leptin signaling in the forebrain. Further studies are needed to directly test this hypothesis.
Assuntos
Metabolismo Energético/efeitos dos fármacos , Hipotálamo/efeitos dos fármacos , Leptina/farmacologia , Fator de Transcrição STAT3/efeitos dos fármacos , Tecido Adiposo Marrom/efeitos dos fármacos , Tecido Adiposo Marrom/metabolismo , Animais , Western Blotting , Relação Dose-Resposta a Droga , Quarto Ventrículo , Hipotálamo/metabolismo , Infusões Intraventriculares , Fígado/efeitos dos fármacos , Fígado/metabolismo , Masculino , Proteína Quinase 1 Ativada por Mitógeno/efeitos dos fármacos , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/efeitos dos fármacos , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Fosfatidilinositol 3-Quinase/efeitos dos fármacos , Fosfatidilinositol 3-Quinase/metabolismo , Fosfatidilinositol 3-Quinases , Fosfoproteínas/efeitos dos fármacos , Fosfoproteínas/metabolismo , Proteína Tirosina Fosfatase não Receptora Tipo 1/efeitos dos fármacos , Proteína Tirosina Fosfatase não Receptora Tipo 1/metabolismo , Ratos , Ratos Sprague-Dawley , Rombencéfalo/efeitos dos fármacos , Rombencéfalo/metabolismo , Fator de Transcrição STAT3/metabolismo , Proteína 3 Supressora da Sinalização de Citocinas/efeitos dos fármacos , Proteína 3 Supressora da Sinalização de Citocinas/metabolismo , Proteína Desacopladora 1/efeitos dos fármacos , Proteína Desacopladora 1/metabolismoRESUMO
Fat transplants increase body fat mass without changing the energy status of an animal and provide a tool for investigating control of total body fat. Early transplant studies found that small pieces of transplanted fat took on the morphology of the transplant recipient. Experiments described here tested whether this response was dependent upon expression of leptin receptors in either transplanted fat or the recipient mouse. Fat from leptin receptor deficient db/db mice or wild-type mice was placed subcutaneously in db/db mice. After 12 wk, cell size distribution in the transplant was the same as in endogenous fat of the recipient. Thus, wild-type fat cells, which express leptin receptors, were enlarged in a hyperleptinemic environment, indicating that leptin does not directly control adipocyte size. By contrast, db/db or wild-type fat transplanted into wild-type mice decreased in size, suggesting that a functional leptin system in the recipient is required for body fat mass to be controlled. In the final experiment, wild-type fat was transplanted into a db/db mouse parabiosed to either another db/db mouse to an ob/ob mouse or in control pairs in which both parabionts were ob/ob mice. Transplants increased in size in db/db-db/db pairs, decreased in db/db-ob/ob pairs and did not change in ob/ob-ob/ob pairs. We propose that leptin from db/db parabionts activated leptin receptors in their ob/ob partners. This, in turn, stimulated release of unidentified circulating factors, which travelled back to the db/db partner and acted on the transplant to reduce fat cell size.
Assuntos
Adipócitos Brancos/metabolismo , Tecido Adiposo Branco/metabolismo , Adiposidade , Leptina/sangue , Adipócitos Brancos/transplante , Tecido Adiposo Branco/transplante , Animais , Biomarcadores/sangue , Tamanho Celular , Feminino , Masculino , Camundongos Knockout , Parabiose , Receptores para Leptina/deficiência , Receptores para Leptina/genética , Transdução de SinaisRESUMO
Leptin receptors (ObRs) in the forebrain and hindbrain have been independently recognized as important mediators of leptin responses. We recently used low-dose leptin infusions to show that chronic activation of both hypothalamic and hindbrain ObRs is required to reduce body fat. The objective of the present study was to identify the brain nuclei that are selectively activated in rats that received chronic infusion of leptin in both the forebrain and hindbrain. Either saline or leptin was infused into third and fourth ventricles (0.1 µg/24 h in the third ventricle and 0.6 µg/24 h in the fourth ventricle) of male Sprague-Dawley rats for 6 days using Alzet pumps. Rats infused with leptin into both ventricles (LL rats) showed a significant increase in phosphorylated (p)STAT3 immunoreactivity in the arcuate nucleus, ventromedial hypothalamus, dorsomedial hypothalamus, and posterior hypothalamus compared with other groups. No differences in pSTAT3 immunoreactivity were observed in midbrain or hindbrain nuclei despite a sixfold higher infusion of leptin into the fourth ventricle than the third ventricle. ΔFosB immunoreactivity, a marker of chronic neuronal activation, showed that multiple brain nuclei were chronically activated due to the process of infusion, but only the arcuate nucleus, ventromedial hypothalamus, dorsomedial hypothalamus, and ventral tuberomamillary nucleus showed a significant increase in LL rats compared with other groups. These data demonstrate that low-dose leptin in the hindbrain increases pSTAT3 in areas of the hypothalamus known to respond to leptin, supporting the hypothesis that leptin-induced weight loss requires an integrated response from both the hindbrain and forebrain.
Assuntos
Hipotálamo/efeitos dos fármacos , Leptina/administração & dosagem , Proteínas Quinases/metabolismo , Rombencéfalo/efeitos dos fármacos , Fator de Transcrição STAT3/metabolismo , Animais , Relação Dose-Resposta a Droga , Hipotálamo/metabolismo , Infusões Intraventriculares , Masculino , Fosforilação/efeitos dos fármacos , Ratos , Ratos Sprague-DawleyRESUMO
We previously reported that a 2-day peripheral infusion of glucosamine caused leptin resistance in rats, suggesting a role for the hexosamine biosynthetic pathway (HBP) in the development of leptin resistance. Here we tested leptin responsiveness in mice in which HBP activity was stimulated by offering 30% sucrose solution in addition to chow and water or by infusing glucosamine. Mice were leptin resistant after 33 days of access to sucrose. Resistance was associated with increased activity of the HBP and with phosphorylation of transcription factor signal transducer and activator of transcription-3 Tyr705 [pSTAT3(Y705)] but inhibition of suppressor of cytokine signaling 3 in the liver and hypothalamus. Intravenous infusion of glucosamine for 3 h stimulated pSTAT3(Y705) but prevented leptin-induced phosphorylation of STAT3(S727). In an in vitro system, glucose, glucosamine, and leptin each dose dependently increased O-linked ß-N-acetylglucosamine (O-GlcNAc) protein and pSTAT3(Y705) in HepG2 cells. To test the effect of glucose on leptin responsiveness cells were incubated in 5.5 mM (LG) or 20 mM (HG) glucose for 18 h and were treated with 0 or 50 ng/ml leptin for 15 min. HG alone and LG + leptin produced similar increases in O-GlcNAc protein, glutamine fructose-6-phosphate amidotransferase (GFAT), and pSTAT3(Y705) compared with LG media. Leptin did not stimulate these proteins in HG cells, suggesting leptin resistance. Leptin-induced pSTAT3(S727) was prevented by HG media. Inhibition of GFAT with azaserine prevented LG + leptin and HG stimulation of pSTAT3. These data demonstrate development of leptin resistance in sucrose-drinking mice and provide new evidence of leptin-induced stimulation of the HBP.
Assuntos
Tronco Encefálico/metabolismo , Sacarose Alimentar/metabolismo , Hexosaminas/biossíntese , Leptina/metabolismo , Fígado/metabolismo , Fator de Transcrição STAT3/metabolismo , Acetilglucosamina/metabolismo , Animais , Glicemia/metabolismo , Sacarose Alimentar/administração & dosagem , Glucosamina/administração & dosagem , Glucosamina/metabolismo , Glutamina-Frutose-6-Fosfato Transaminase (Isomerizante)/metabolismo , Células Hep G2 , Humanos , Infusões Intravenosas , Insulina/sangue , Leptina/sangue , Masculino , Camundongos Endogâmicos C57BL , Fosforilação , Fator de Transcrição STAT3/sangue , Transdução de Sinais , Fatores de TempoRESUMO
Rats offered 30% sucrose solution in addition to chow and water become leptin resistant therefore we investigated the effect of sucrose solution consumption on leptin signaling. In Experiment 1 rats were resistant to 3rd ventricle injections of1.5 µg leptin after 36 days of sucrose and western blot indicated that resistance was associated with increased basal levels of signal transducer and activator of transcription 3 phosphorylation (pSTAT3). In Experiment 2 rats were resistant to a peripheral injection of 2mg leptin/kg after 26 days of sucrose. Immunohistochemistry indicated that increased basal pSTAT3 was limited to the medial and lateral arcuate nucleus of the hypothalamus. Increased availability of glucose and fructose can stimulate the hexosamine biosynthetic pathway (HBP) which O-GlcNAc-modifies proteins. This has the potential to change protein bioactivity. We tested whether this pathway could account for the leptin resistance. There was no increase in the expression of HBP enzymes in tissues from sucrose rats in Experiment 1, however, direct activation of the HBP with a 3h intravenous infusion of 30 µmol/kg/min glucosamine significantly increased hypothalamic pSTAT3. Although sucrose consumption and activation of the HBP both increase hypothalamic pSTAT3 experiments described here did not provide evidence of a direct link between sucrose consumption, HBP activity and leptin resistance. Unexpectedly, we found that the HBP enzyme glutamine fructose-6-phosphate amidotransferase (GFAT) in liver and O-GlcNAcase in hypothalamus were increased 30min after leptin injection in leptin responsive animals, implying a complex interaction between activity of the HBP and leptin responsiveness.
Assuntos
Sacarose Alimentar/administração & dosagem , Hexosaminas/metabolismo , Leptina/metabolismo , Acetilglucosamina/metabolismo , Animais , Vias Biossintéticas/efeitos dos fármacos , Glicemia/efeitos dos fármacos , Fármacos do Sistema Nervoso Central/farmacologia , Ingestão de Alimentos/efeitos dos fármacos , Ingestão de Alimentos/fisiologia , Glucosamina/farmacologia , Glutamina-Frutose-6-Fosfato Transaminase (Isomerizante)/metabolismo , Hipotálamo/efeitos dos fármacos , Hipotálamo/metabolismo , Insulina/sangue , Fígado/enzimologia , Masculino , N-Acetilglucosaminiltransferases/metabolismo , Fosforilação/efeitos dos fármacos , Ratos Sprague-Dawley , Rombencéfalo/efeitos dos fármacos , Rombencéfalo/metabolismo , Fator de Transcrição STAT3/metabolismoRESUMO
Stress activates multiple neural and endocrine systems to allow an animal to respond to and survive in a threatening environment. The corticotropin-releasing factor system is a primary initiator of this integrated response, which includes activation of the sympathetic nervous system and the hypothalamic-pituitary-adrenal (HPA) axis. The energetic response to acute stress is determined by the nature and severity of the stressor, but a typical response to an acute stressor is inhibition of food intake, increased heat production, and increased activity with sustained changes in body weight, behavior, and HPA reactivity. The effect of chronic psychological stress is more variable. In humans, chronic stress may cause weight gain in restrained eaters who show increased HPA reactivity to acute stress. This phenotype is difficult to replicate in rodent models where chronic psychological stress is more likely to cause weight loss than weight gain. An exception may be hamsters subjected to repeated bouts of social defeat or foot shock, but the data are limited. Recent reports on the food intake and body composition of subordinate members of group-housed female monkeys indicate that these animals have a similar phenotype to human stress-induced eaters, but there are a limited number of investigators with access to the model. Few stress experiments focus on energy balance, but more information on the phenotype of both humans and animal models during and after exposure to acute or chronic stress may provide novel insight into mechanisms that normally control body weight.