RESUMO
The effects of continuous or intermittent access to a high-caloric (HC) diet, always offered in addition to standard chow, on body mass and leptin resistance were analyzed in female C57BL/6J mice. Susceptibility for diet-induced obesity (DIO) was apparent from the marked preference for the HC diet. Continuous HC diet feeding of mice at 4 weeks of age induced leptin resistance within 2 weeks and massive gains in body mass, although with increasing inter-individual variability in the inbred strain considered to be isogenic. In adult mice receiving HC diet for the first time, leptin treatment failed to reduce energy intake first after 11 days of HC diet feeding, but became effective again within 3 days after HC diet withdrawal. In mice with a history of several preceding periods of access to the HC diet totalling >30 days, supplementary HC diet abolished the anorectic effect of leptin treatment within only 3 days and it reappeared not earlier than 11 days after HC diet withdrawal. Thus, in the investigated DIO-prone mouse strain both, the loss of responsiveness to leptin under HC diet and its recovery after HC diet withdrawal strongly depended on the dietary history. Recovery from leptin resistance during periods of intermittent chow feeding was associated with losses of body mass that did not completely compensate for the obesity-inducing effect of the preceding HC diet.
Assuntos
Gorduras na Dieta/administração & dosagem , Ingestão de Alimentos/fisiologia , Leptina/administração & dosagem , Periodicidade , Fatores Etários , Animais , Comportamento Animal , Índice de Massa Corporal , Peso Corporal/fisiologia , Resistência a Medicamentos , Ingestão de Energia/fisiologia , Feminino , Leptina/efeitos adversos , Camundongos , Camundongos Endogâmicos C57BLRESUMO
We tried to unravel why leptin's fat store depleting action levels off in the course of long-term applications. Supplying leptin by minipump infusion for 2 months to ob/ob mice at rates between 115 pmol day(-1) and 460 pmol day(-1) resulted in stable plasma leptin levels between 0.2 ng ml(-1) and 8 ng ml(-1). Initial treatment effects were leptin dose-dependent reductions in food intake and body mass, especially in fat content, followed by re-increases of food intake to levels only 4-18% below pre-treatment levels. Decreased body mass subsequently stabilized dose-dependently with body fat contents between 4% and 33% showing that total fat depletion was not a precondition for the progressive reduction of leptin-induced anorexia. Oxygen consumption measurements excluded contributions of enhanced energy dissipation to fat depletion. Plasma insulin concentrations declined from excessively high pre-treatment levels to steady, leptin dose-dependent levels within the normal range. Temporary anorexia in response to repeated additional 1-day leptin injections (100 pmol g(-1) day(-1)) remained unchanged throughout long-term leptin infusion. Among various alternatives considered to explain the adipostatic equilibrium attained at new, dose-dependent levels under long-term leptin treatment, interaction between the leptin signal and at least one counteracting signal increasing with fat depletion is proposed as the most plausible working hypothesis.
Assuntos
Tecido Adiposo/efeitos dos fármacos , Leptina/administração & dosagem , Obesidade/tratamento farmacológico , Tecido Adiposo/patologia , Tecido Adiposo/fisiopatologia , Animais , Composição Corporal/efeitos dos fármacos , Relação Dose-Resposta a Droga , Resistência a Medicamentos , Ingestão de Alimentos/efeitos dos fármacos , Metabolismo Energético/efeitos dos fármacos , Retroalimentação , Insulina/sangue , Leptina/sangue , Masculino , Camundongos , Camundongos Obesos , Obesidade/patologia , Obesidade/fisiopatologia , Transdução de Sinais , Redução de Peso/efeitos dos fármacosRESUMO
Body weight regulation is mediated through several major signaling pathways, some of which have been delineated by positional cloning of spontaneous genetic mutations in mice. Lepr(db/db) mice are obese due to a defect in the signaling portion of the leptin receptor, which has led to extensive study of this highly conserved system over the past several years. We have created an allelic series at Lepr for the further examination of LEPR signaling phenotypes using both the FLP /frt and CRE /loxP systems. By inserting a frt-PGK-neo-frt sequence in Lepr intron 16, we have generated a conditional gene repair Lepr allele ( Lepr-neo) that elicits morbid obesity, diabetes, and infertility in homozygous mice, recapitulating the obesity syndrome of Lepr(db/db) mice. Thus, in vivo excision of the PGK-neo cassette with a FLP recombinase transgene restores the lean and fertile phenotype to Lepr(flox/flox) mice. In the same construct, we have also inserted loxP sites that flank Lepr coding exon 17, a region that encodes a JAK docking site required for STAT3 signaling. CRE-mediated excision of Lepr coding exon 17 from Lepr with a frameshift in subsequent exons results in a syndrome of obesity, diabetes, and infertility in LeprDelta17/Delta17 mice, which is indistinguishable from Lepr(neo/neo) and Lepr(db/db) mice. We conclude that suppression of Lepr gene expression by PGK-neo is phenotypically equivalent to deletion of the Lepr signaling motifs, and therefore the Lepr(neo/neo) mouse may be used to investigate conditional gene repair of Lepr signaling deficiency.
Assuntos
Alelos , DNA Nucleotidiltransferases/genética , Integrases/genética , Obesidade/genética , Receptores de Superfície Celular/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Glicemia/metabolismo , Regulação da Temperatura Corporal/genética , Peso Corporal/genética , Peso Corporal/fisiologia , Quimera , Temperatura Baixa , Modelos Animais de Doenças , Ingestão de Alimentos/genética , Ingestão de Alimentos/fisiologia , Feminino , Insulina/sangue , Masculino , Camundongos , Dados de Sequência Molecular , Obesidade/metabolismo , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Receptores de Superfície Celular/biossíntese , Receptores para Leptina , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
In young (35- to 56-day-old) and middle-aged (9-mo-old) wild-type (+/+) and melanocortin-4 receptor (MC4R)-deficient (+/-, -/-) mice, expressions of neuropeptide Y (NPY), agouti-related protein (AGRP), pro-opiomelanocortin (POMC), and cocaine-and-amphetamine-regulated transcript (CART) were analyzed in the arcuate nucleus (ARC) and adjacent regions comprising the dorsomedial (DMN) and ventromedial (VMN) nucleus. In the ARC of young mice, NPY and AGRP expression increased and POMC and CART expression decreased with body fat content. Adjusting for the influence of body fat content by ANCOVA showed that the levels of NPY, POMC, and CART were highest and of AGRP lowest in young -/- mice. In the middle-aged mice, feedback from body fat content was weakened. For -/- mice ANCOVA revealed higher NPY and AGRP, lower POMC, and unchanged CART expression levels relative to young -/- mice. In the DMN and VMN, POMC and AGRP signals were absent at each age. CART was expressed in the DMN independent of age, fat content, and genotype. For NPY expression, an age-dependent induction was found in the DMN and VMN; it was absent in the young but present in the middle-aged mice, showing close positive correlations between body fat content and the numbers of NPY-labeled cells which were further enhanced in -/- mice. Thus MC4R deficiency augments age-induced NPY expression in the DMN and VMN with no feedback from body fat content. Negative feedback control by body fat content on ARC neuropeptide expression is present in young animals but vanishes with age and is modulated by MC4R deficiency.
Assuntos
Tecido Adiposo/metabolismo , Envelhecimento/metabolismo , Deleção de Genes , Hipotálamo/metabolismo , Neuropeptídeos/metabolismo , Receptor Tipo 4 de Melanocortina/deficiência , Receptor Tipo 4 de Melanocortina/metabolismo , Proteína Relacionada com Agouti , Animais , Núcleo Arqueado do Hipotálamo/metabolismo , Genótipo , Peptídeos e Proteínas de Sinalização Intercelular , Leptina/sangue , Camundongos , Camundongos Knockout , Modelos Animais , Proteínas do Tecido Nervoso/análise , Proteínas do Tecido Nervoso/metabolismo , Neuropeptídeo Y/análise , Neuropeptídeo Y/metabolismo , Obesidade/sangue , Obesidade/genética , Obesidade/metabolismo , Pró-Opiomelanocortina/análise , Pró-Opiomelanocortina/metabolismo , Proteínas/análise , Proteínas/metabolismo , Receptor Tipo 4 de Melanocortina/genéticaRESUMO
Previous studies on mice with melanocortin-4 receptor gene (MC4r) knockout have focused on obese adults. Because humans with functional MC4r mutations show early-onset obesity, we determined the onset of excessive fat deposition in 10- to 56-day-old mice, taking into account sex and litter influences. Total body fat content of MC4r-/- on day 35 and MC4r+/- on day 56 significantly exceeds that of MC4r+/+. Plasma leptin levels increase in proportion to fat mass. According to cumulative food intake and energy expenditure measurements from day 21 to 35, onset of excessive fat deposition in MC4r-/- is fueled by hyperphagia and counteracted partially by hypermetabolism. In 35- to 56-day-old mice, arcuate nucleus neuropeptide Y (NPY) mRNA decreases and pro-opiomelanocortin (POMC) mRNA increases with fat content and plasma leptin levels independently of genotype. Taking into account fat content by ANCOVA reveals, however, increases in both NPY mRNA and POMC mRNA due to melanocortin-4 receptor (MC4R) deficiency. We conclude that hyperphagia, not hypometabolism, is the primary disturbance initiating excessive fat deposition in MC4R-deficient mice at weaning and that the overall changes in NPY and POMC expression tend to antagonize the onset of excessive fat deposition.
Assuntos
Metabolismo Energético/genética , Hiperfagia/genética , Obesidade/genética , Receptores da Corticotropina/deficiência , Receptores da Corticotropina/genética , Tecido Adiposo/crescimento & desenvolvimento , Animais , Composição Corporal/genética , Ingestão de Alimentos/genética , Ingestão de Energia/genética , Feminino , Genótipo , Hiperfagia/sangue , Hiperfagia/fisiopatologia , Leptina/sangue , Masculino , Camundongos , Camundongos Knockout , Neuropeptídeo Y/biossíntese , Obesidade/sangue , Obesidade/fisiopatologia , Fragmentos de Peptídeos/biossíntese , Pró-Opiomelanocortina/biossíntese , Receptor Tipo 4 de Melanocortina , Receptores da Corticotropina/fisiologiaRESUMO
In small mammals leptin reduces fat stores not only by inhibiting food intake but also by disinhibiting metabolic cold defense. Presuming that postnatal age and feeding regime set the conditions for either mode of leptin action, we compared the caloric equivalents of changes in fat mass, metabolic rate (MR) and food intake (FI) induced by 10-day treatments with leptin, in rats treated from postnatal days 7, 15, 25 onward and in adult mice that were free-feeding or food restricted. Whereas MR changes are known to dominate from postnatal days 7-16, changes in MR explained only about 50% and 30% of the leptin-induced changes in fat mass between days 15-24 and days 25-34, respectively. In adult mice of similar body weights, leptin-induced reductions in fat mass under free-feeding conditions were due only to FI decreases but due only to MR increases under food-restricted conditions. Thus, the same leptin treatment induces the same percentage decrease in body fat content by driving the two effectors differently, depending on age and feeding conditions. Consequently, in assessing the effects of leptin under various physiological conditions, short-term measurements of FI or MR alone are not sufficient. Instead, determination of the resultant decreases in total body fat is required.
Assuntos
Envelhecimento/metabolismo , Ração Animal , Animais Recém-Nascidos/metabolismo , Leptina/farmacologia , Tecido Adiposo/anatomia & histologia , Criação de Animais Domésticos , Animais , Animais Recém-Nascidos/crescimento & desenvolvimento , Animais Lactentes/metabolismo , Composição Corporal/efeitos dos fármacos , Ingestão de Alimentos , Feminino , Privação de Alimentos/fisiologia , Masculino , Camundongos , Camundongos Endogâmicos , Tamanho do Órgão , Ratos , Ratos Zucker , DesmameRESUMO
To compare the anorectic effectiveness of leptin and the amylin analogue salmon calcitonin (sCT), rodents were treated on 1 day with subcutaneous injections. In chow-fed C57Bl/6J mice, leptin and sCT reduced energy intake and acted additively. After C57Bl/6J mice had become leptin-resistant on being fed chocolate as a palatable high-caloric supplement to chow, their sCT-induced decrease in energy intake was more pronounced than in chow-fed mice with differential changes in the intake of chocolate (strong reduction) and chow (slight increase). Dose-response relationships for sCT-induced reductions in energy intake were analysed in chow-fed C57Bl/6J mice and two obese strains, ob/ob mice and melanocortin-4 receptor knockout (MC4-r-KO) mice, as well as in wild-type and fatty (fa/fa) rats. Compared to C57Bl/6J mice, reduction in food intake induced by sCT was attenuated in MC4-r-KO mice, and nearly absent in ob/ob mice, over the dose range investigated. Compared to C57Bl/6J mice, wild-type rats responded more sensitively to sCT and its efficiency was only slightly reduced in fatty (fa/fa) rats. Thus, while genetically induced failures of leptin signalling reduce the action of sCT, it effectively inhibits the intake of a palatable, high fat-high sugar diet even in states of diet-induced obesity with functional leptin resistance.
Assuntos
Calcitonina/farmacologia , Ingestão de Alimentos/efeitos dos fármacos , Leptina/fisiologia , Transdução de Sinais/fisiologia , Animais , Índice de Massa Corporal , Dieta , Ingestão de Energia/efeitos dos fármacos , Feminino , Insulina/sangue , Leptina/deficiência , Leptina/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos Obesos , Ratos , Ratos Endogâmicos , Receptor Tipo 4 de Melanocortina , Receptores da Corticotropina/deficiência , Receptores da Corticotropina/genética , Receptores para LeptinaRESUMO
The homeostatic regulation of body weight protects the organism from the negative consequences of starvation and obesity. Glucocorticoids (GCs) modulate this regulation, although the underlying mechanisms remain unclear. To address the role of central GRs in the regulation of energy balance, we studied mice in which GRs have selectively been inactivated in the nervous system. Mutant mice display marked growth retardation. During suckling age this is associated with normal fat deposition causing a 60% temporary increase of percent body fat, compared with control littermates. After weaning, fat and protein depositions are reduced so that adults are both smaller and leaner than their controls. Decreased food intake and, after weaning, reduced metabolic efficiency account for these developmental disturbances. Plasma levels of leptin and insulin, two important energy balance regulators, are elevated in young mutants but normal in adults. Leptin/body fat ratio is higher at all ages, suggesting disturbed control of circulating leptin as a consequence of chronically elevated GC levels in mutant animals. Adult mutants display increased hypothalamic CRH and NPY levels, but peptide levels of melanin concentrating hormone and Orexin A and B are unchanged. The increased levels of plasma GCs and hypothalamic CRH may act as catabolic signals most likely leading to persistently reduced energy accumulation.