RESUMO
AIMS: Epinephrine is unique among biogenic catecholamines as a potent agonist of ß2-adrenoceptors. The ß2-adrenoceptor mediated effects during development might be linked to the increase of epinephrine synthesis. Our purpose was to characterize ß-adrenoceptor-mediated relaxation in the aorta of newborn and young rabbits (3 to 4months old), and to relate those responses with the epinephrine content of the adrenal gland. MAIN METHODS: The epinephrine levels and the tyrosine hydroxylase activity were determined in adrenal glands of newborn and young rabbits. Also, concentration-response curves to phenylephrine (selective α1-adrenoceptor agonist), dobutamine (selective ß1-adrenoceptor agonist), terbutaline (selective ß2-adrenoceptor agonist), and CL 316243 (selective ß3-adrenoceptor agonist) were determined in isolated aortic rings obtained from both groups. KEY FINDINGS: The adrenal gland content and the plasma concentrations of epinephrine were lower in newborn than in young rabbits. In contrast, the tyrosine hydroxylase activity was higher in newborn than in young rabbits. On the other hand, the maximal response to phenylephrine was lower in newborn than in young rabbits. Terbutaline at concentrations selective for ß2-adrenoceptors had no relaxing effects in neonates, in contrast to young rabbits. The potency and the maximal response of neither dobutamine nor CL 316243 were significantly different between the two groups. SIGNIFICANCE: In rabbits, as well as in humans, ß2-adrenoceptor-mediated responses and epinephrine synthesis are both immature at birth. On the other hand, the ß1 and ß3-adrenoceptor-mediated responses are fully developed. We conclude that epinephrine may influence the development of the ß2-adrenoceptor-mediated responses at birth and the rabbit is an excellent model to study these issues.
Assuntos
Epinefrina/metabolismo , Receptores Adrenérgicos beta/deficiência , Vasodilatação , Glândulas Suprarrenais/efeitos dos fármacos , Glândulas Suprarrenais/metabolismo , Agonistas alfa-Adrenérgicos/farmacologia , Animais , Animais Recém-Nascidos , Dobutamina/farmacologia , Epinefrina/sangue , Norepinefrina/sangue , Norepinefrina/metabolismo , Fenilefrina/farmacologia , Coelhos , Receptores Adrenérgicos alfa/metabolismo , Receptores Adrenérgicos beta/metabolismo , Terbutalina/farmacologia , Tirosina 3-Mono-Oxigenase/metabolismo , Vasodilatação/efeitos dos fármacosRESUMO
BACKGROUND AND PURPOSE: Adenylyl cyclase (AC) is a key signalling enzyme for many GPCRs and catalyses the conversion of ATP to cAMP which, in turn, is a crucial determinant of many biological responses. ß-Adrenoceptor agonists are prescribed as bronchodilators for asthma and chronic obstructive pulmonary disease, and it is commonly assumed that they elicit their actions via AC-dependent production of cAMP. However, empirical evidence in support of this is lacking and the exact mechanism by which these drugs acts remains elusive. This is partly due to the existence of at least 10 different isoforms of AC and the absence of any truly selective pharmacological inhibitors. Here, we have used genetically modified mice and model systems to establish the role of AC isoforms in the airway responses to ß-adrenoceptor agonists. EXPERIMENTAL APPROACH: Receptors mediating responses to ß-adrenoceptor agonists in airway smooth muscle (ASM) and sensory nerve were identified in isolated tissue systems. Expression of mRNA for the AC isoforms in ASM and neurones was determined by qPCR. Functional responses were assessed in AC isoform KO mice and wild-type controls. KEY RESULTS: Airway and vagal tissue expressed mRNA for various isoforms of AC. AC6 was the most prominent isoform. Responses to ß-adrenoceptor agonists in tissues from AC6 KO mice were virtually abolished. CONCLUSIONS AND IMPLICATIONS: AC6 played a critical role in relaxation of ASM to ß1 -adrenoceptor agonists and in modulation of sensory nerves by ß1-3 -adrenoceptor agonists. These results further unravel the signalling pathway of this extensively prescribed class of medicine.
Assuntos
Adenilil Ciclases/fisiologia , Músculo Liso/fisiologia , Receptores Adrenérgicos beta/fisiologia , Traqueia/fisiologia , Nervo Vago/fisiologia , Adenilil Ciclases/deficiência , Adenilil Ciclases/genética , Agonistas Adrenérgicos beta/farmacologia , Antagonistas Adrenérgicos beta/farmacologia , Animais , Dinoprostona/análogos & derivados , Dinoprostona/farmacologia , Etanolaminas/farmacologia , Fenoterol/farmacologia , Regulação Enzimológica da Expressão Gênica , Cobaias , Imidazóis/farmacologia , Técnicas In Vitro , Isoenzimas/genética , Masculino , Camundongos Knockout , Contração Muscular/efeitos dos fármacos , Músculo Liso/efeitos dos fármacos , Propanolaminas/farmacologia , Receptores Adrenérgicos beta/deficiência , Receptores Adrenérgicos beta/genética , Receptores de Prostaglandina E Subtipo EP2/agonistas , Transdução de Sinais , Traqueia/efeitos dos fármacos , Nervo Vago/efeitos dos fármacosRESUMO
The melanocortin receptor 4 (MC4R) plays a major role in body weight regulation and its agonist MTII has been widely used to study the role of MC4Rs in energy expenditure promotion and feeding reduction. Unexpectedly, we observed that intraperitoneal (i.p.) administration of MTII induced a rapid reduction in both body temperature and energy expenditure, which was independent of its effect on feeding and followed by a prolonged increase in energy expenditure. The rapid reduction was at least partly mediated by brain neurons since intracerebroventricular (icv) administration of alpha melanocyte-stimulating hormone, an endogenous melanocortin receptor agonist, produced a similar response. In addition, the body temperature-lowering effect of MTII was independent of the presence of MC4Rs, but in a similar fashion to the previously shown effect on body temperature by 5'AMP. Moreover, ß-adrenergic receptors (ß-ARs) were required for the recovery from low body temperature induced by MTII and further pharmacological studies showed that the MTII's effect on body temperature may be partially mediated by the vasopressin V1a receptors. Collectively, our results reveal a previously unappreciated role for the melanocortin pathway in rapidly lowering body temperature.
Assuntos
Temperatura Corporal/efeitos dos fármacos , Peptídeos Cíclicos/farmacologia , Receptor Tipo 4 de Melanocortina/agonistas , alfa-MSH/análogos & derivados , Animais , Temperatura Corporal/fisiologia , Metabolismo Energético/efeitos dos fármacos , Metabolismo Energético/fisiologia , Camundongos , Camundongos Knockout , Receptor Tipo 4 de Melanocortina/deficiência , Receptor Tipo 4 de Melanocortina/fisiologia , Receptores Adrenérgicos beta/deficiência , Receptores Adrenérgicos beta/genética , Receptores Adrenérgicos beta/fisiologia , Receptores de Vasopressinas/fisiologia , alfa-MSH/farmacologiaRESUMO
Acute exposure to particulate matter (PM) air pollution causes thrombotic cardiovascular events, leading to increased mortality rates; however, the link between PM and cardiovascular dysfunction is not completely understood. We have previously shown that the release of IL-6 from alveolar macrophages is required for a prothrombotic state and acceleration of thrombosis following exposure to PM. Here, we determined that PM exposure results in the systemic release of catecholamines, which engage the ß2-adrenergic receptor (ß2AR) on murine alveolar macrophages and augment the release of IL-6. In mice, ß2AR signaling promoted the development of a prothrombotic state that was sufficient to accelerate arterial thrombosis. In primary human alveolar macrophages, administration of a ß2AR agonist augmented IL-6 release, while the addition of a beta blocker inhibited PM-induced IL-6 release. Genetic loss or pharmacologic inhibition of the ß2AR on murine alveolar macrophages attenuated PM-induced IL-6 release and prothrombotic state. Furthermore, exogenous ß2AR agonist therapy further augmented these responses in alveolar macrophages through generation of mitochondrial ROS and subsequent increase of adenylyl cyclase activity. Together, these results link the activation of the sympathetic nervous system by ß2AR signaling with metabolism, lung inflammation, and an enhanced susceptibility to thrombotic cardiovascular events.
Assuntos
Agonistas de Receptores Adrenérgicos beta 2/administração & dosagem , Agonistas de Receptores Adrenérgicos beta 2/efeitos adversos , Interleucina-6/biossíntese , Material Particulado/administração & dosagem , Material Particulado/efeitos adversos , Trombose/etiologia , Adenilil Ciclases/biossíntese , Animais , Antitrombina III/biossíntese , Líquido da Lavagem Broncoalveolar/química , Catecolaminas/biossíntese , Colforsina/administração & dosagem , Humanos , Macrófagos Alveolares/efeitos dos fármacos , Macrófagos Alveolares/fisiologia , Masculino , Camundongos , Camundongos Knockout , Peptídeo Hidrolases/biossíntese , Propranolol/administração & dosagem , Propranolol/efeitos adversos , Espécies Reativas de Oxigênio/metabolismo , Receptores Adrenérgicos beta/deficiência , Receptores Adrenérgicos beta/genética , Receptores Adrenérgicos beta 1/deficiência , Receptores Adrenérgicos beta 1/genéticaRESUMO
We previously showed that leucine deprivation decreases abdominal fat mass largely by increasing energy expenditure, as demonstrated by increased lipolysis in white adipose tissue (WAT) and uncoupling protein 1 (UCP1) expression in brown adipose tissue (BAT). The goal of the present study was to investigate the possible involvement of central nervous system (CNS) in this regulation and elucidate underlying molecular mechanisms. For this purpose, levels of genes and proteins related to lipolysis in WAT and UCP1 expression in BAT were analyzed in wild-type mice after intracerebroventricular administration of leucine or corticotrophin-releasing hormone antibodies, or in mice deleted for three ß-adrenergic receptors, after being maintained on a leucine-deficient diet for 7 d. Here, we show that intracerebroventricular administration of leucine significantly attenuates abdominal fat loss and blocks activation of hormone sensitive lipase in WAT and induction of UCP1 in BAT in leucine-deprived mice. Furthermore, we provide evidence that leucine deprivation stimulates fat loss by increasing expression of corticotrophin-releasing hormone in the hypothalamus via activation of stimulatory G protein/cAMP/protein kinase A/cAMP response element-binding protein pathway. Finally, we show that the effect of leucine deprivation on fat loss is mediated by activation of the sympathetic nervous system. These results suggest that CNS plays an important role in regulating fat loss under leucine deprivation and thereby provide novel and important insights concerning the importance of CNS leucine in the regulation of energy homeostasis.
Assuntos
Adiposidade , Hormônio Liberador da Corticotropina/metabolismo , Hipotálamo/metabolismo , Leucina/deficiência , Sistema Nervoso Simpático/metabolismo , Gordura Abdominal/efeitos dos fármacos , Gordura Abdominal/metabolismo , Tecido Adiposo Marrom/metabolismo , Adiposidade/efeitos dos fármacos , Animais , Anticorpos/administração & dosagem , Anticorpos/farmacologia , Hormônio Liberador da Corticotropina/imunologia , AMP Cíclico/metabolismo , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Metabolismo Energético/efeitos dos fármacos , Subunidades alfa Gs de Proteínas de Ligação ao GTP/metabolismo , Hipotálamo/efeitos dos fármacos , Hipotálamo/enzimologia , Injeções Intraventriculares , Leucina/administração & dosagem , Leucina/farmacologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Ratos , Ratos Sprague-Dawley , Receptores Adrenérgicos beta/deficiência , Receptores Adrenérgicos beta/metabolismo , Transdução de Sinais/efeitos dos fármacos , Sistema Nervoso Simpático/efeitos dos fármacosRESUMO
We investigated a possible role of the central glucagon-like peptide (GLP-1) receptor system as an essential brain circuit regulating adiposity through effects on nutrient partitioning and lipid metabolism independent from feeding behavior. Both lean and diet-induced obesity mice were used for our experiments. GLP-1 (7-36) amide was infused in the brain for 2 or 7 d. The expression of key enzymes involved in lipid metabolism was measured by real-time PCR or Western blot. To test the hypothesis that the sympathetic nervous system may be responsible for informing adipocytes about changes in CNS GLP-1 tone, we have performed direct recording of sympathetic nerve activity combined with experiments in genetically manipulated mice lacking beta-adrenergic receptors. Intracerebroventricular infusion of GLP-1 in mice directly and potently decreases lipid storage in white adipose tissue. These effects are independent from nutrient intake. Such CNS control of adipocyte metabolism was found to depend partially on a functional sympathetic nervous system. Furthermore, the effects of CNS GLP-1 on adipocyte metabolism were blunted in diet-induced obese mice. The CNS GLP-1 system decreases fat storage via direct modulation of adipocyte metabolism. This CNS GLP-1 control of adipocyte lipid metabolism appears to be mediated at least in part by the sympathetic nervous system and is independent of parallel changes in food intake and body weight. Importantly, the CNS GLP-1 system loses the capacity to modulate adipocyte metabolism in obese states, suggesting an obesity-induced adipocyte resistance to CNS GLP-1.
Assuntos
Sistema Nervoso Central/metabolismo , Metabolismo dos Lipídeos/fisiologia , Obesidade/fisiopatologia , Receptores de Glucagon/fisiologia , Transdução de Sinais/fisiologia , Sistema Nervoso Simpático/fisiologia , Tecido Adiposo/citologia , Tecido Adiposo/metabolismo , Análise de Variância , Animais , Composição Corporal/efeitos dos fármacos , Composição Corporal/genética , Composição Corporal/fisiologia , Sistema Nervoso Central/efeitos dos fármacos , Ingestão de Alimentos/efeitos dos fármacos , Metabolismo Energético/efeitos dos fármacos , Metabolismo Energético/fisiologia , Peptídeo 1 Semelhante ao Glucagon/farmacologia , Receptor do Peptídeo Semelhante ao Glucagon 1 , Metabolismo dos Lipídeos/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Atividade Motora/efeitos dos fármacos , Atividade Motora/fisiologia , Obesidade/etiologia , Obesidade/metabolismo , Fragmentos de Peptídeos/farmacologia , Receptores Adrenérgicos beta/deficiência , Receptores de Glucagon/antagonistas & inibidores , Transdução de Sinais/genética , Sistema Nervoso Simpático/efeitos dos fármacos , Fatores de TempoRESUMO
Mitochondrial uncoupling protein 1 (UCP1) is a key regulator of adaptive thermogenesis and energy expenditure. Mice lacking UCP1 are cold sensitive, but surprisingly not obese at room temperature. In this issue of Cell Metabolism, Feldmann et al. (2009) unmask an obesogenic phenotype by simply maintaining these mice at thermoneutrality.
Assuntos
Metabolismo Energético , Canais Iônicos/genética , Proteínas Mitocondriais/genética , Termogênese , Tecido Adiposo Marrom/metabolismo , Animais , Diabetes Mellitus/prevenção & controle , Humanos , Camundongos , Camundongos Knockout , Obesidade/prevenção & controle , Receptores Adrenérgicos beta/deficiência , Temperatura , Proteína Desacopladora 1RESUMO
Activation of beta2-adrenergic receptors inhibits osteoblastic bone formation and enhances osteoclastic bone resorption. Whether beta-blockers inhibit ovariectomy-induced bone loss and decrease fracture risk remains controversial. To further explore the role of beta-adrenergic signaling in skeletal acquisition and response to estrogen deficiency, we evaluated mice lacking the three known beta-adrenergic receptors (beta-less). Body weight, percent fat, and bone mineral density were significantly higher in male beta-less than wild-type (WT) mice, more so with increasing age. Consistent with their greater fat mass, serum leptin was significantly higher in beta-less than WT mice. Mid-femoral cross-sectional area and cortical thickness were significantly higher in adult beta-less than WT mice, as were femoral biomechanical properties (+28 to +49%, P < 0.01). Young male beta-less had higher vertebral (1.3-fold) and distal femoral (3.5-fold) trabecular bone volume than WT (P < 0.001 for both) and lower osteoclast surface. With aging, these differences lessened, with histological evidence of increased osteoclast surface and decreased bone formation rate at the distal femur in beta-less vs. WT mice. Serum tartrate-resistance alkaline phosphatase-5B was elevated in beta-less compared with WT mice from 8-16 wk of age (P < 0.01). Ovariectomy inhibited bone mass gain and decreased trabecular bone volume/total volume similarly in beta-less and WT mice. Altogether, these data indicate that absence of beta-adrenergic signaling results in obesity and increased cortical bone mass in males but does not prevent deleterious effects of estrogen deficiency on trabecular bone microarchitecture. Our findings also suggest direct positive effects of weight and/or leptin on bone turnover and cortical bone structure, independent of adrenergic signaling.
Assuntos
Densidade Óssea/genética , Ovariectomia/efeitos adversos , Receptores Adrenérgicos beta/deficiência , Envelhecimento/fisiologia , Animais , Fenômenos Biomecânicos , Composição Corporal/fisiologia , Feminino , Fêmur/anatomia & histologia , Fêmur/diagnóstico por imagem , Fêmur/fisiologia , Vértebras Lombares/diagnóstico por imagem , Masculino , Camundongos , Camundongos Knockout , Coluna Vertebral/anatomia & histologia , Coluna Vertebral/fisiologia , Tomografia Computadorizada por Raios XRESUMO
It is generally accepted that cardiac sympathetic tone dominates the control of heart rate (HR) in mice. However, we have recently challenged this notion given that HR in the mouse is responsive to ambient temperature (T(a)) and that the housing T(a) is typically 21-23 degrees C, well below the thermoneutral zone ( approximately 30 degrees C) of this species. To specifically test the hypothesis that cardiac sympathetic tone is the primary mediator of HR control in the mouse, we first examined the metabolic and cardiovascular responses to rapid changes in T(a) to demonstrate the sensitivity of the mouse cardiovascular system to T(a). We then determined HR in 1) mice deficient in cardiac sympathetic tone ("beta-less" mice), 2) mice deficient in cardiac vagal tone [muscarinic M(2) receptor (M(2)R(-/-)) mice], and 3) littermate controls. At a T(a) of 30 degrees C, the HR of beta-less mice was identical to that of wild-type mice (351 +/- 11 and 363 +/- 10 beats/min, respectively). However, the HR of M(2)R(-/-) mice was significantly greater (416 +/- 7 beats/min), demonstrating that vagal tone predominates over HR control at this T(a). When these mice were calorically restricted to 70% of normal intake, HR fell equally in wild-type, beta-less, and M(2)R(-/-) mice (DeltaHR = 73 +/- 9, 76 +/- 3, and 73 +/- 7 beats/min, respectively), suggesting that the fall in intrinsic HR governs bradycardia of calorically restricted mice. Only when the T(a) was relatively cool, at 23 degrees C, did beta-less mice exhibit a HR (442 +/- 14 beats/min) that was different from that of littermate controls (604 +/- 10 beats/min) and M(2)R(-/-) mice (602 +/- 5 beats/min). These experiments conclusively demonstrate that in the absence of cold stress, regulation of vagal tone and modulation of intrinsic rate are important determinants of HR control in the mouse.
Assuntos
Sistema Nervoso Autônomo/metabolismo , Temperatura Corporal , Frequência Cardíaca , Coração/inervação , Receptor Muscarínico M2/metabolismo , Receptores Adrenérgicos beta/metabolismo , Nervo Vago/metabolismo , Animais , Pressão Sanguínea , Bradicardia/etiologia , Bradicardia/metabolismo , Bradicardia/fisiopatologia , Restrição Calórica/efeitos adversos , Temperatura Baixa , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Receptor Muscarínico M2/deficiência , Receptor Muscarínico M2/genética , Receptores Adrenérgicos beta/deficiência , Receptores Adrenérgicos beta/genéticaRESUMO
OBJECTIVE: To investigate the impact of beta-adrenoceptor deficiency on the metabolic effects of leptin. MEASUREMENTS: Leptin was infused subcutaneously through an osmotic minipump in wild-type (WT) and beta(1)/beta(2)/beta(3)-adrenoceptor knockout (beta-less) mice and its effects on food intake, energy expenditure, carbohydrate and lipid utilization as well as on the levels of expression of the brown adipose tissue (BAT), thermogenic marker uncoupling protein-1 (UCP1) and type II deiodinase (D2) mRNAs were compared. RESULTS: Leptin treatment decreased food intake by 23% in both the WT and the beta-less mice. In pair-fed animals being used as controls, leptin treatment was found to increase energy expenditure in WT, but not in beta-less mice. No difference was observed in carbohydrate or fat utilization between leptin-treated WT and beta-less mice. Leptin increased UCP1 and D2 mRNA levels in WT mouse BAT 1.7- and 3-fold, respectively, but had no effect on the expression of these genes in beta-less mouse BAT. CONCLUSION: The stimulatory effects of leptin on oxygen consumption, BAT UCP1 and D2 expression require functional beta-adrenoceptors, but its inhibitory effect on food intake and its stimulatory effect on fat utilization is independent of the beta-adrenoceptor signalling.
Assuntos
Tecido Adiposo Marrom/metabolismo , Ingestão de Energia/efeitos dos fármacos , Metabolismo Energético/efeitos dos fármacos , Leptina/farmacologia , Receptores Adrenérgicos beta/deficiência , Tecido Adiposo Marrom/efeitos dos fármacos , Animais , Calorimetria Indireta/métodos , Métodos de Alimentação , Iodeto Peroxidase/metabolismo , Canais Iônicos/metabolismo , Masculino , Camundongos , Camundongos Knockout , Proteínas Mitocondriais/metabolismo , Reação em Cadeia da Polimerase , RNA Mensageiro/metabolismo , Receptores Adrenérgicos beta/fisiologia , Proteína Desacopladora 1 , Iodotironina Desiodinase Tipo IIRESUMO
We and others have previously shown that triple knockout mice lacking the beta1/beta2/beta3-adrenoceptors (beta-less mice) developed a progressive obesity at adulthood. Here, we studied the glucose homeostasis in beta-less mice before the onset of obesity. We show that beta-less mice have increased fat mass and are glucose intolerant. In addition, we observed that beta-less mice have impaired glucose-induced insulin secretion and exhibit an increase in liver PEPCK gene expression in the fed state, suggesting that they have increased gluconeogenesis. Although these characteristics are usually associated with insulin resistance, beta-less mice exhibit enhanced insulin sensitivity during insulin tolerance tests. This is keeping with the results obtained during euglycemic-hyperinsulinemic clamps showing that beta-less mice display increased insulin responsiveness with normal suppression of hepatic glucose production. Altogether, our results suggest that an intact beta-adrenergic system is required to regulate overall glucose homeostasis and, in particular, insulin-mediated glucose uptake, most likely at the level of muscles and adipose tissue.
Assuntos
Intolerância à Glucose/fisiopatologia , Insulina/fisiologia , Receptores Adrenérgicos beta/deficiência , Aumento de Peso , Animais , Glicemia/metabolismo , Composição Corporal , Ingestão de Alimentos , Regulação Enzimológica da Expressão Gênica , Gluconeogênese , Glucose/metabolismo , Técnica Clamp de Glucose , Homeostase , Insulina/sangue , Insulina/farmacologia , Cinética , Camundongos , Camundongos Knockout , Proteínas Serina-Treonina Quinases/genética , Proteínas Serina-Treonina Quinases/metabolismoRESUMO
Hyperthyroidism and states of adrenergic hyperactivity have many common clinical features, suggesting similar pathogenic mechanisms of action. The widespread use of beta-adrenergic receptor (betaAR) antagonists (beta-blockers) to treat hyperthyroidism has led to the belief that the physiological consequences of thyroid hormone (TH) excess are mediated in part via catecholamine signaling through betaARs. To test this hypothesis, we compared the response to TH excess in mice lacking the three known betaARs (beta-less) vs. wild-type (WT) mice. Although beta-less mice had a lower heart rate at baseline in comparison to WT mice, the metabolic and cardiovascular responses to hyperthyroidism were equivalent in both WT and beta-less mice. These data indicate that the metabolic and cardiovascular effects of TH excess are largely independent of betaARs. These findings suggest that the efficacy of clinical treatment of hyperthyroidism with beta-blockers is due to antagonism of sympathetic signaling, and that this process functions independently of TH action.
Assuntos
Sistema Cardiovascular/fisiopatologia , Hipertireoidismo/fisiopatologia , Receptores Adrenérgicos beta/metabolismo , Agonistas Adrenérgicos beta/farmacologia , Animais , Hipertireoidismo/induzido quimicamente , Hipertireoidismo/metabolismo , Isoproterenol/farmacologia , Camundongos , Receptores Adrenérgicos beta/deficiência , Tri-IodotironinaRESUMO
Obesity is characterized by an excessive development of fat mass which is a consequence of increased fat cell size and/or fat cell number. Several hormones and neurotransmitters are regulators of adipose tissue development and metabolism. Among them, catecholamines play a major role by acting through alpha 2- and beta-adrenergic receptors. Stimulation of alpha 2-adrenergic receptors induce inhibition of lipolysis in mature adipocytes as well as preadipocyte proliferation. The antilipolytic effect mediated by alpha 2-adrenergic receptors is in part responsible for the weak lipid mobilization of some fat deposits in humans (subcutaneous fat in particular). Changes in beta- and alpha 2-adrenergic receptors ratio and function have been proposed to explain the lipolytic disturbances described in some obese subjects. Human and rodent adipocytes differ considerably with respect to the balance between beta- and alpha 2-adrenergic receptors. Human adipocytes express mainly alpha 2- but very few beta 3-adrenergic receptors while the reverse is true for rodent adipocytes. Since no suitable animal model was available to study the contribution of alpha 2/beta-adrenergic balance in adipocytes in vivo, we combined gene targeting and transgenic approaches to create a mice with increased alpha 2/beta-adrenergic ratio in adipose tissue. Specifically, we have generated transgenic mice strains on a beta 3-adrenergic receptor knock-out background which express human alpha 2-adrenergic receptors. No particular phenotype was observed in mice maintained in normal diet whereas when fed a high fat diet, transgenic mice increased significantly body weight and fat mass. These results underline the physiologic relevance of the interaction of the presence of alpha 2-adrenergic receptors with a high fat diet in the control of adipose tissue development.
Assuntos
Obesidade/genética , Receptores Adrenérgicos alfa/fisiologia , Receptores Adrenérgicos beta/fisiologia , Animais , Humanos , Camundongos , Camundongos Knockout , Camundongos Transgênicos , Obesidade/fisiopatologia , Receptores Adrenérgicos alfa/deficiência , Receptores Adrenérgicos beta/deficiênciaRESUMO
Administration of beta-adrenergic receptor (beta-AR) agonists, especially beta(3)-AR agonists, is well known to increase thermogenesis in rodents and humans. In this work we studied the role of the beta(3)-AR in regulating mRNA expression of genes involved in thermogenesis, i.e., mitochondrial uncoupling proteins UCP2 and UCP3, and peroxisome proliferator-activated receptor-gamma coactivator-1 (PGC-1), in mouse skeletal muscle. For this purpose, different beta(3)-AR agonists were administered acutely to both wild type mice and mice whose beta(3)-AR gene has been disrupted (beta(3)-AR KO mice). CL 316243 increased the expression of UCP2, UCP3 and PGC-1 in wild type mice only. By contrast, BRL 37344 and CGP 12177 increased the expression of UCP2 and UCP3 in both wild type and beta(3)-AR KO mice, whereas they increased the expression of PGC-1 in wild type mice only. Finally, acute (3 h) cold exposure increased the expression of UCP2 and UCP3, but not PGC-1, in skeletal muscle of both wild type and beta(3)-AR KO mice. These results show that selective stimulation of the beta(3)-AR affects the expression of UCP2, UCP3 and PGC-1 in skeletal muscle. This effect is probably indirect, as muscle does not seem to express beta(3)-AR. In addition, our data suggest that BRL 37344 and CGP 12177 act, in part, through an as yet unidentified receptor, possibly a beta(4)-AR.
Assuntos
Expressão Gênica , Proteínas de Membrana Transportadoras , Proteínas Mitocondriais , Receptores Adrenérgicos beta/fisiologia , Fatores de Transcrição/genética , Desacopladores , Agonistas Adrenérgicos beta/farmacologia , Animais , Glicemia/metabolismo , Proteínas de Transporte/genética , Temperatura Baixa , Ácidos Graxos não Esterificados/sangue , Feminino , Canais Iônicos , Camundongos , Camundongos Knockout , Músculo Esquelético/metabolismo , Proteínas/genética , RNA Mensageiro/metabolismo , Receptores Adrenérgicos beta/deficiência , Receptores Adrenérgicos beta/genética , Proteína Desacopladora 2 , Proteína Desacopladora 3RESUMO
Over the past twenty years, several lines of evidence from preclinical and clinical studies has accumulated suggesting that a GABA deficit may be involved in mood disorders, particularly in depression, and that increasing GABAergic neurotransmission may exert an antidepressant effect and perhaps a mood stabilizing effect. Given that GABA has an inhibitory effect on biogenic amine neurotransmitters such as norepinephrine and serotonin and this inhibition may be involved in local circuits and interneurons, it has been suggested that the hypothesis of a GABA deficit in mood disorders does not compete with but complements the well-established hypotheses of alterations in noradrenergic and serotonergic function in mood disorders. In this paper, we systematically reviewed the results from preclinical and clinical studies of GABA function in the pathophysiology of mood disorders and in the mechanism of action of mood stabilizers, antidepressants and electroconvulsive therapy. We also discussed the unifying theory of the neurochemistry of mood disorders, which integrates the GABA hypothesis into the biogenic amine hypotheses, and indicated future directions for research.
Assuntos
Transtornos do Humor/fisiopatologia , Ácido gama-Aminobutírico/fisiologia , Animais , Antidepressivos/uso terapêutico , Antimaníacos/uso terapêutico , Eletroconvulsoterapia , Humanos , Transtornos do Humor/terapia , Receptores Adrenérgicos beta/deficiência , Receptores Adrenérgicos beta/fisiologia , Receptores de GABA-A/deficiência , Receptores de GABA-A/fisiologiaRESUMO
Some blockers of beta1- and beta2-adrenoceptors cause cardiostimulant effects through an atypical beta-adrenoceptor (putative beta4-adrenoceptor) that resembles the beta3-adrenoceptor. It is likely but not proven that the putative beta4-adrenoceptor is genetically distinct from the beta3-adrenoceptor. We therefore investigated whether or not the cardiac atypical beta-adrenoceptor could mediate agonist effects in mice lacking a functional beta3-adrenoceptor gene (beta3 KO). (-)-CGP 12177, a beta1- and beta2-adrenoceptor blocker that causes agonist effects through both beta3-adrenoceptors and cardiac putative beta4-adrenoceptors, caused cardiostimulant effects that were not different in atria from wild-type (WT) mice and beta3 KO mice. The effects of (-)-CGP 12177 were resistant to blockade by (-)-propranolol (200 nM) but were blocked by (-)-bupranolol (1 microM) with an equilibrium dissociation constant of 15 nM in WT and 17 nM in beta3 KO. (-)-[3H]CGP 12177 labeled a similar density of the putative beta4-adrenoceptor in ventricular membranes from the hearts of both WT (Bmax = 52 fmol/mg protein) and beta3 KO (Bmax = 53 fmol/mg protein) mice. The affinity of (-)-[3H]CGP 12177 for the cardiac putative beta4-adrenoceptor was not different between WT (Kd = 46 nM) and beta3 KO (Kd= 40 nM). These results provide definitive evidence that the cardiac putative beta4-adrenoceptor is distinct from the beta3-adrenoceptor.
Assuntos
Agonistas Adrenérgicos beta/metabolismo , Cardiotônicos/farmacologia , Miocárdio/metabolismo , Propanolaminas/farmacologia , Receptores Adrenérgicos beta/deficiência , Receptores Adrenérgicos beta/genética , Animais , Sítios de Ligação/genética , Southern Blotting , Feminino , Genótipo , Átrios do Coração/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Mutagênese Insercional , Propanolaminas/metabolismo , Receptores Adrenérgicos beta 3RESUMO
Adipocytes from genetically obese (ob/ob) mice display an impaired response to beta-adrenergic stimulation, but the molecular defects have not been unequivocally identified. The expression and functional activity of the beta 1-, beta 2-, and beta 3-adrenergic receptor (AR) subtypes in white and brown adipose tissue from genetically lean and obese (ob/ob) mice were compared. Three beta 3AR transcripts of 2.1, 2.6, and 3.5 kilobases were identified in adipose tissue from lean mice by Northern blotting. All three beta 3AR mRNA species were dramatically reduced (by approximately 300-fold) in 12-week-old obese mice compared to those in lean animals. beta 1AR mRNA levels were also reduced (by approximately 4-fold) in obese mice, whereas beta 2AR mRNA levels were not significantly changed. The functional consequences of these changes in beta 3AR and beta 1AR expression were assessed by measuring beta-agonist-stimulated adenylyl cyclase activity in adipocyte plasma membranes with subtype-selective beta-adrenergic agonists and antagonists. Dose-response curves with epinephrine from lean mice were best fit to a two-component model comprised of 23% high affinity (K(act) = 1.42 x 10(-7) M) and 77% low affinity (K(act) = 1.67 x 10(-5) M) components, corresponding to activation of beta 1AR and beta 2AR conjointly, and beta 3AR, respectively. The beta 1AR-selective antagonist CGP20712A reduced the high affinity component to about 10%, whereas the nonselective beta-antagonist propranolol eliminated the high affinity component. The beta 3AR-selective agonist BRL37344 stimulated adenylyl cyclase activity in lean membranes to a slightly lesser extent than epinephrine, but was more potent (73% high affinity component; K(act) = 3.61 x 10(-8) M). In obese mice, stimulation of adenylyl cyclase by all agonists was severely blunted and was best fit to a single class of sites. Studies with CGP20712A or the beta 2AR-selective antagonist ICI118,551 indicated that this residual response was predominantly beta 2AR in character. Expression of beta AR subtypes in both brown and white adipose tissue of weanling obese mice (4-5-weeks of age) was also affected, but to a lesser extent, consistent with the progressive severity of obesity with age. Together the reduction in expression of the beta 3AR and beta 1AR impairs the beta-agonist-stimulated adenylyl cyclase response over a broad concentration range by greatly lowering the maximum stimulation and shifting the adrenergic sensitivity at low concentrations from a mixed beta 1AR/beta 2AR response to predominantly beta 2AR.
Assuntos
Tecido Adiposo/metabolismo , Agonistas Adrenérgicos beta/farmacologia , Antagonistas Adrenérgicos beta/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Hiperglicemia/metabolismo , Camundongos Mutantes/metabolismo , Obesidade/metabolismo , Receptores Adrenérgicos beta/deficiência , Adenilil Ciclases/efeitos dos fármacos , Adenilil Ciclases/metabolismo , Tecido Adiposo/efeitos dos fármacos , Tecido Adiposo Marrom/efeitos dos fármacos , Tecido Adiposo Marrom/metabolismo , Animais , Sequência de Bases , Células Cultivadas , AMP Cíclico/fisiologia , Diabetes Mellitus Tipo 2 , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Ativação Enzimática/efeitos dos fármacos , Epinefrina/farmacologia , Hiperglicemia/genética , Imidazóis/farmacologia , Lipólise/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Mutantes/genética , Dados de Sequência Molecular , Obesidade/congênito , Obesidade/genética , Propanolaminas/farmacologia , Propranolol/farmacologia , Splicing de RNA , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Receptores Adrenérgicos beta/classificação , Receptores Adrenérgicos beta/efeitos dos fármacos , Receptores Adrenérgicos beta/genética , Receptores Adrenérgicos beta/fisiologiaRESUMO
The aim of this study was to investigate the relative numbers of myocardial beta 1 and beta 2 receptors in pigs. Membrane particles from left ventricular porcine and mixed ventricular rat myocardium were examined for subtypes of beta adrenergic receptors with a radioligand binding technique using [125I]-cyanopindolol (ICYP) as trace, and the new highly beta 1 selective antagonist Sandoz 204 545 and the beta 2 selective antagonist ICI 118 551 for displacement. Radioligand displacement experiments were also performed using propranolol, isoprenaline and terbutaline. The displacement curves obtained with the subtype selective antagonists and agonist revealed biphasic inhibition of specific ICYP binding in rat preparations, indicating a beta 1/beta 2 ratio of approximately 2/1. In porcine preparations displacement of specific ICYP binding with all agents resulted in monophasic curves, thus sharply contrasting the rat preparations. Affinity constants of displacing drugs derived from these monophasic curves indicated that the specific binding site was a beta 1 receptor. No displacement compatible with beta 2 affinity was found. In the same rat preparations we found that adenylate cyclase activation and inhibition by beta receptor subtype specific agonists and antagonists were mediated by two receptor subtypes, whereas in the pig, adenylate cyclase activation and its inhibition seemed to occur via only one receptor subtype, the beta 1 adrenoceptor.
Assuntos
Miocárdio/análise , Receptores Adrenérgicos beta/análise , Suínos/metabolismo , Adenilil Ciclases/metabolismo , Agonistas Adrenérgicos beta/metabolismo , Agonistas Adrenérgicos beta/farmacologia , Antagonistas Adrenérgicos beta/metabolismo , Antagonistas Adrenérgicos beta/farmacologia , Animais , Ligação Competitiva , Ativação Enzimática/efeitos dos fármacos , Ventrículos do Coração , Ratos , Ratos Endogâmicos/metabolismo , Receptores Adrenérgicos beta/classificação , Receptores Adrenérgicos beta/deficiênciaRESUMO
In order to raise efficacy of primary prevention of platinosis an obsidan-based test (40 mg for internal use) was also used in vocational selection. After the obsidan-based test the decrease of pneumotachometric indicators was noted that reflected deficiency of the functional state of bronchial beta-adrenoreceptors by 6% and higher and was regarded as unfavourable. Of 110 examined persons 16 (14.5%) had significant test results. Similar deficiency was detected in 27 workers with respiratory platinosis, that fact verified the significance of beta-AR deficiency in the disease pathogenesis. The possibility of raising efficacy of primary prevention of respiratory platinosis was stressed due to the use of the additional obsidan-based test.
Assuntos
Doenças Profissionais/prevenção & controle , Gestão de Recursos Humanos , Seleção de Pessoal , Platina/efeitos adversos , Doenças Respiratórias/prevenção & controle , Adulto , Brônquios/inervação , Feminino , Humanos , Masculino , Doenças Profissionais/induzido quimicamente , Propranolol , Receptores Adrenérgicos beta/deficiência , Doenças Respiratórias/induzido quimicamenteRESUMO
The number of beta-receptors was determined in 60 atopic patients: 30 asthmatics with sensitization to domestic inhalant allergens and 30 rhinitic patients with sensitization to pollen and/or Dermatophagoides antigens. A L-iodocyanopindolol radio-active marker was used. Our control population (20 healthy individuals) showed 908 +/- 396 receptors/cell. The rhinitic individuals showed a 24% decrease in receptors in relation to the control population. On the other hand, within the asthmatic patients there existed 2 differentiated groups: inactive asthmatics whose receptors were found to be within the normal limits and active asthmatics with a 36% decrease in receptors in relation to the healthy individuals (673 receptors/cell) establishing a significant statistical difference (p less than 0.005). As a consequence these data seem to support Szentivanyi's B-adrenoreceptor theory.