RESUMO
In this brief review we summarize the current fndings relative to the discovery of a small peptide ligand, phoenixin (PNX). Using a bioinformatic approach, two novel peptides PNX-14 and PNX-20 containing 14 and 20 amino acids, respectively, were isolated from diverse tissues including the brain, heart, lung and stomach. Mass spectrometry analysis identified a major and minor peak corresponding to PNX-14 and PNX-20, in rat or mouse spinal cord extracts. With the use of a rabbit polyclonal antiserum, phoenixin immunoreactivity (irPNX) was detected in discrete areas of the rodent brain including several hypothalamic subnuclei and dorsal motor nucleus of the vagus. In addition, irPNX was detected in a population of sensory ganglion cells including dorsal root ganglion, nodose ganglion and trigeminal ganglion, and in cell processes densely distributed to the superficial layers of the dorsal horn, nucleus of the solitary tract and spinal trigeminal tract. irPNX cell processes were also detected in the skin and myenteric plexus, suggesting a brain-gut and/or brain-skin connection. Pharmacological studies show that PNX-14 injected subcutaneously to the nape of the neck of mice provoked dose-dependent repetitive scratching bouts directed to the back of the neck with the hindpaws. Our result suggests that the peptide PNX-14 and/or PNX-20, may serve as one of the endogenous signal molecules transducing itch sensation. Additionally, results from other laboratories show that exogenous PNX may affect a number of diverse behaviors such as memory formation, depression, reproduction, food-intake and anxiolytic-like behaviors.
Assuntos
Hormônios Hipotalâmicos/fisiologia , Hormônios Peptídicos/fisiologia , Peptídeos/fisiologia , Sequência de Aminoácidos , Animais , Humanos , Hormônios Hipotalâmicos/administração & dosagem , Hormônios Hipotalâmicos/química , Hipotálamo/metabolismo , Memória/fisiologia , Plexo Mientérico/metabolismo , Hormônios Peptídicos/administração & dosagem , Hormônios Peptídicos/química , Peptídeos/administração & dosagem , Peptídeos/química , Prurido/metabolismo , Medula Espinal/metabolismoRESUMO
Nesfatin-1 is a recently identified satiety molecule detectable in neurons of the hypothalamus and nucleus of solitary tract (NTS). Immunohistochemical studies revealed nesfatin-1-immunoreactive (irNEF) cells in the Edinger-Westphal nucleus, dorsal motor nucleus of vagus, and caudal raphe nuclei of the rats, in addition to the hypothalamus and NTS reported in the initial study. Double-labeling immunohistochemistry showed that irNEF cells were vasopressin or oxytocin positive in the paraventricular and supraoptic nucleus; cocaine-amphetamine-regulated transcript or tyrosine hydroxylase positive in arcuate nucleus; cocaine-amphetamine-regulated transcript or melanin concentrating hormone positive in the lateral hypothalamus. In the brainstem, irNEF neurons were choline acetyltransferase positive in the Edinger-Westphal nucleus and dorsal motor nucleus of vagus; tyrosine hydroxylase positive in the NTS; and 5-hydroxytryptamine positive in the caudal raphe nucleus. The biological activity of rat nesfatin-1 (1-82) (100 nm) was assessed by the Ca(2+) microfluorometric method. Nesfatin-1 elevated intracellular Ca(2+) concentrations [Ca(2+)](i) in dissociated and cultured hypothalamic neurons. The response was prevented by pretreating the cells with pertussis toxin (100 nm) or Ca(2+)-free solution and by a combination of the L-type and P/Q-type calcium channel blocker verapamil (1 microm) and omega-conotoxin MVIIC (100 nm). The protein kinase A inhibitor KT 5720 (1 microm) suppressed nesfatin-1-induced rise in [Ca(2+)](i). The result shows that irNEF is distributed to several discrete nuclei in the brainstem, in addition to the hypothalamus and NTS reported earlier, and that the peptide interacts with a G protein-coupled receptor, leading to an increase of [Ca(2+)](i), which is linked to protein kinase A activation in cultured rat hypothalamic neurons.
Assuntos
Proteínas do Tecido Nervoso/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Animais , Cálcio/metabolismo , Proteínas de Ligação ao Cálcio , Células Cultivadas , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Proteínas de Ligação a DNA , Ativação Enzimática , Feminino , Hipotálamo/citologia , Hipotálamo/metabolismo , Imuno-Histoquímica , Membranas Intracelulares/metabolismo , Masculino , Bulbo/metabolismo , Neurônios/metabolismo , Nucleobindinas , Concentração Osmolar , Ponte/metabolismo , Ratos , Ratos Sprague-Dawley , Distribuição TecidualRESUMO
With the use of an antiserum directed against the human smooth muscle-associated protein 8 (SMAP8) fragment SMAP8(98-138), Western blot and immunohistochemical studies revealed SMAP8 expression in the rat brain. A band with a molecular size of about 45 kDa was detected in tissues from the rat hypothalamus and a weaker band from the cortex. SMAP8 immunoreactivity (irSMAP8) was detected in neurons of the hypothalamic paraventricular, supraoptic, and supraoptic retrochiasmatic nuclei; a few irSMAP8 cells were scattered in the zona incerta as well as the cerebral cortex. Immunoreactive cell processes were detected mostly in the internal layer of the median eminence. Double labeling the hypothalamic sections with SMAP8 and vasopressin (VP) or oxytocin (OT) antiserum revealed that a population of VP- and OT-immunoreactive neurons expressed irSMAP8. The biological activity of SMAP8 in rat central neurons was assessed by the calcium microfluorimetric Fura-2 method. SMAP8 (100 nM) elevated cytosolic calcium concentrations [Ca2+]i in a population of dissociated and cultured rat hypothalamic neurons; the response was eliminated in Ca2+-free saline. This is the first evidence of irSMAP8 in a population of VP/OT-containing hypothalamic neurons in the rat, and the peptide is biologically active in hypothalamic neurons, as evidenced by mobilization of extracellular Ca2+.
Assuntos
Encéfalo/metabolismo , Proteínas Musculares/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Animais , Encéfalo/anatomia & histologia , Cálcio/metabolismo , Células Cultivadas , Humanos , Hipotálamo/metabolismo , Imuno-Histoquímica , Masculino , Neurônios/metabolismo , Ocitocina/metabolismo , Ratos , Ratos Sprague-Dawley , Vasopressinas/metabolismoRESUMO
Cocaine- and amphetamine-regulated transcript (CART) peptide consists of a family of peptides. Expression of the peptide fragment CART(1-39) was explored in the rat using an antiserum directed against CART(1-39) of the short form of the human CART prohormone. CART(1-39)-immunoreactivity, herein referred to as irCART, was detected in the rat central and peripheral nervous tissues with a pattern similar to that labeled with the antiserum CART(55-102) or CART(79-102). For example, irCART cells were detected in the hypothalamus, pons, medulla oblongata, spinal cord, and adrenal medulla. In urethane-anesthetized rats, CART(1-39) (0.05 to 2 nmol) by intrathecal injection did not cause a significant change of blood pressure or heart rate, but potentiated the pressor effects of glutamate injected intrathecally. Lastly, the effect of CART(1-39) on intracellular calcium concentrations [Ca2+]i was assessed and compared to that caused by CART(55-102) in cultured rat cortical neurons using the microfluorimetric method. CART(1-39) (100 nM) induced two types of responses in a population of cortical neurons: 1) a slowly rising increase in [Ca2+]i superimposed with oscillations, and 2) a fast increase followed by a sustained increase of [Ca2+]i. CART(55-102) caused only a slowly rising increase in [Ca2+]i in cortical neurons. Our result shows that the expression pattern of irCART in the rat nervous system and the potentiating action of CART(1-39) on glutamate-induced pressor response is similar to that reported for CART(55-102); but the calcium mobilizing action of CART(1-39) differs from that of CART(55-102), suggesting the possible existence of multiple CART receptors coupled to different calcium signaling pathways.
Assuntos
Proteínas do Tecido Nervoso/metabolismo , Glândulas Suprarrenais/metabolismo , Animais , Pressão Sanguínea/efeitos dos fármacos , Tronco Encefálico/metabolismo , Cálcio/metabolismo , Células Cultivadas , Gânglios/metabolismo , Ácido Glutâmico/administração & dosagem , Frequência Cardíaca/efeitos dos fármacos , Humanos , Hipotálamo/metabolismo , Imuno-Histoquímica , Injeções Espinhais , Líquido Intracelular/metabolismo , Masculino , Proteínas do Tecido Nervoso/administração & dosagem , Proteínas do Tecido Nervoso/química , Neurônios/citologia , Neurônios/metabolismo , Fragmentos de Peptídeos/metabolismo , Ratos , Ratos Sprague-Dawley , Medula Espinal/metabolismo , Sistema Nervoso Simpático/metabolismoRESUMO
Intermedin is a novel member of the calcitonin/calcitonin gene-related peptide (CGRP) family peptide, which has vasodilatory and hypotensive actions identical to those of adrenomedullin and CGRP. Cleavage sites located between 2 basic amino acids at Arg93-Arg94 result in the production of prepro-intermedin95-147, namely intermedin1-53. The bioactive action of intermedin1-53 and its physiological significance are unclear. In this work, we aimed to explore the effects of intermedin1-53 on acute myocardial injury induced by isoproterenol. Myocardial ischemia injury in rats was induced by subcutaneous injection of a high dose of isoproterenol, and the therapeutic effect of intermedin1-53 was observed. Plasma lactate dehydrogenase activity, myocardial and plasma malondialdehyde content were higher in the isoproterenol group than that in controls. Isoproterenol-treated rats showed lower maximal rate of increase and decrease of left-ventricle pressure development (+/-left-ventricle dp/dtmax) and higher left-ventricle end-diastolic pressure (all P<0.01), which suggested severe heart failure and myocardial injury. Semi-quantitative RT-PCR analysis showed that the gene expression of calcitonin receptor-like receptor and receptor-activity-modifying protein (RAMP)1, RAMP2 and RAMP3 in ventricular myocardia were up-regulated by 79% (P<0.01), 48% (P<0.01), 31% (P<0.05) and 130% (P<0.01), respectively, compared with controls. In myocardial sarcolemmal membranes, the maximum binding capacity for [125I]-intermedin1-53 was increased by 118% (P<0.01) in the isoproterenol group compared with controls. Rats treated with low dosage intermedin1-53 (5 nmol/kg/day, 2 days) showed 21% (P<0.05) higher myocardial cAMP content, 18% and 31% higher+left-ventricle dp/dtmax and -left-ventricle dp/dtmax respectively, 288% lower left-ventricle end-diastolic pressure (all P<0.01), and attenuated myocardial lactate dehydrogenase leakage and malondialdehyde formation (all P<0.01). Treatment with high dosage intermedin1-53 (20 nmol/kg/day, 2 days) gave better results than that with low dosage intermedin1-53. These results suggest that the intermedin receptor system was up-regulated in isoproterenol-induced myocardial ischemic injury and intermedin1-53 might play a pivotal cardioprotective role in such injury.
Assuntos
Adrenomedulina/farmacologia , Isoproterenol/toxicidade , Infarto do Miocárdio/prevenção & controle , Neuropeptídeos/farmacologia , Fragmentos de Peptídeos/farmacologia , Adrenomedulina/química , Adrenomedulina/metabolismo , Animais , Proteína Semelhante a Receptor de Calcitonina , AMP Cíclico/metabolismo , Relação Dose-Resposta a Droga , Expressão Gênica/efeitos dos fármacos , Injeções Subcutâneas , Peptídeos e Proteínas de Sinalização Intracelular/genética , Radioisótopos do Iodo , Isoproterenol/administração & dosagem , L-Lactato Desidrogenase/sangue , Masculino , Malondialdeído/sangue , Malondialdeído/metabolismo , Proteínas de Membrana/genética , Infarto do Miocárdio/induzido quimicamente , Infarto do Miocárdio/fisiopatologia , Miocárdio/metabolismo , Miocárdio/patologia , Neuropeptídeos/química , Neuropeptídeos/metabolismo , Fragmentos de Peptídeos/metabolismo , Ligação Proteica/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Proteína 2 Modificadora da Atividade de Receptores , Proteína 3 Modificadora da Atividade de Receptores , Proteínas Modificadoras da Atividade de Receptores , Receptores da Calcitonina/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sarcolema/efeitos dos fármacos , Sarcolema/metabolismoRESUMO
Insulin-like peptide 5 (INSL5) mRNA was detected in the mouse hypothalamus by RT-PCR. Immunohistochemical studies using an antiserum against the mouse INSL5 peptide revealed INSL5-immunoreactive (irINSL5) neurons in the paraventricular, supraoptic, accessory secretory, and supraoptic retrochiasmatic nuclei and immunoreactive cell processes in the internal layer of the median eminence. In the pituitary, irINSL5 was detected in terminal-like elements of the posterior lobe and in cells of the anterior lobe. Double-labeling experiments showed that irINSL5 is expressed in vasopressin-, but not oxytocin-containing neurons. INSL5 (100 nm) administered to dissociated and cultured mouse hypothalamic neurons elevated cytosolic calcium concentrations [Ca(2+)](i), as assessed by the microfluorimetric fura-2 method. In a Ca(2+)-free medium, INSL5 induced in dissociated neurons an increase of [Ca(2+)](i), which was sensitive to the endoplasmic reticulum calcium pump inhibitor thapsigargin (1 microm) and the IP(3) receptor blocker 2-aminoethoxydiphenyl borate (100 microm) or xestospongin C (5 microm). Our result provides the first evidence that INSL5 is expressed in a population of cells in the mouse hypothalamus and pituitary and that it elevates [Ca(2+)](i) by a mechanism involving both Ca(2+) influx and Ca(2+) release from intracellular stores. The concentration of irINSL5 in the hypothalamic-pituitary axis suggests a neuroendocrine function of this insulin superfamily member.
Assuntos
Encéfalo/metabolismo , Cálcio/metabolismo , Insulina/biossíntese , Animais , Células Cultivadas , Citosol/metabolismo , Hipotálamo/metabolismo , Inositol 1,4,5-Trifosfato/metabolismo , Camundongos , Camundongos Endogâmicos ICR , Microscopia Confocal , Neurônios/metabolismo , Hipófise/metabolismo , Proteínas , RNA Mensageiro/metabolismoRESUMO
Electroacupuncture (EA) has been reported to reduce body weight in overweight subjects in clinical practice, as well as in rats and mice with diet-induced obesity. In the present study, this effect of EA was tested in lean rats subjected to long-term food restriction (FR, food was offered only 1 h/day). Two hertz EA administered once every other day produced a further reduction in body weight in FR rats. Exploration of the mechanism involved revealed significant downregulation of the orexigenic peptides: ghrelin in the stomach, and neuropeptide Y (NPY) but not Agouti-related peptide (AgRP) in the hypothalamus, which was in line with the reduction in food intake in rats receiving EA stimulation as compared with those receiving restraint only. Uncoupling protein 3 (UCP3), involved in accelerating energy expenditure, was not significantly altered. These results suggest that the EA-induced body weight reduction was due mainly to a decrease in food intake rather than an increase in energy expenditure. A reduction in the orexigenic peptides ghrelin and NPY may be involved in the underlying mechanism.
Assuntos
Eletroacupuntura , Mucosa Gástrica/metabolismo , Hipotálamo/metabolismo , Neuropeptídeo Y/metabolismo , Hormônios Peptídicos/metabolismo , Proteína Relacionada com Agouti , Animais , Glicemia/análise , Peso Corporal/fisiologia , Ingestão de Alimentos/fisiologia , Privação de Alimentos , Grelina , Hipotálamo/citologia , Imuno-Histoquímica , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Canais Iônicos/metabolismo , Masculino , Proteínas Mitocondriais/metabolismo , Músculo Esquelético/citologia , Músculo Esquelético/metabolismo , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Estômago/citologia , Triglicerídeos/análise , Triglicerídeos/sangue , Proteína Desacopladora 3RESUMO
Beacon is a novel peptide isolated from the hypothalamus of Israeli sand rat. In the present study, we determined the distribution of beacon in the rat brain using immunohistochemical approach with a polyclonal antiserum directed against the synthetic C-terminal peptide fragment (47-73). The hypothalamus represented the major site of beacon-immunoreactive (IR) cell bodies that were concentrated in the paraventricular nucleus (PVN) and the supraoptic nucleus (SON). Additional immunostained cells were found in the septum, bed nucleus of the stria terminalis, subfornical organ and subcommissural organ. Beacon-IR fibers were seen with high density in the internal layer of the median eminence and low to moderate density in the external layer. Significant beacon-IR fibers were also seen in the nucleus of the solitary tract and lateral reticular formation. The beacon neurons found in the PVN were further characterized by double label immunohistochemistry. Several beacon-IR neurons that resided in the medial PVN were shown to coexpress corticotrophin-releasing hormone (CRH) and most labeled beacon fibers in the external layer of median eminence coexist with CRH. The topographical distribution of beacon-IR in the brain suggests multiple biological activities for beacon in addition to its proposed roles in modulating feeding behaviors and pituitary hormone release.
Assuntos
Encéfalo/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Ubiquitinas/metabolismo , Animais , Encéfalo/irrigação sanguínea , Encéfalo/imunologia , Tronco Encefálico/imunologia , Tronco Encefálico/metabolismo , Hormônio Liberador da Corticotropina/biossíntese , Hormônio Liberador da Corticotropina/imunologia , Hormônio Liberador da Corticotropina/metabolismo , Diencéfalo/imunologia , Diencéfalo/metabolismo , Feminino , Glicilglicina/química , Hipotálamo/imunologia , Hipotálamo/metabolismo , Imuno-Histoquímica , Masculino , Proteínas do Tecido Nervoso/biossíntese , Proteínas do Tecido Nervoso/imunologia , Neuropeptídeos/biossíntese , Neuropeptídeos/imunologia , Neuropeptídeos/metabolismo , Sistemas Neurossecretores/imunologia , Sistemas Neurossecretores/metabolismo , Sistemas Neurossecretores/fisiologia , Neurotransmissores/biossíntese , Neurotransmissores/imunologia , Neurotransmissores/metabolismo , Núcleo Hipotalâmico Paraventricular/imunologia , Núcleo Hipotalâmico Paraventricular/metabolismo , Ratos , Ratos Sprague-Dawley , Órgão Subfornical/imunologia , Órgão Subfornical/metabolismo , Telencéfalo/imunologia , Telencéfalo/metabolismo , Ubiquitinas/biossíntese , Ubiquitinas/imunologiaRESUMO
It was reported that acupuncture or electro-acupuncture (EA) is effective in reducing the body weight for obese patients, although the mechanisms remain obscure. In a previous study, we have found that rats fed with high-fat (HIF) diet developed diet-induced obesity (DIO) with a concomitant decrease in the hypothalamic content of the cocaine and amphetamine-regulated transcript (CART) peptide, a peptide with anorexiogenic effect. To assess the central effect of EA on DIO rat, we revealed that EA up-regulated the expression of CART peptide in the arcuate nucleus (ARC) of the DIO rats. After feeding with HIF diet for 14 weeks, the DIO rats received EA stimulation three times per week for 4 weeks. The expression of CART peptide in ARC was measured using immunohistochemistry. The plasma ACTH was measured with ELISA. EA caused a reduction of both body weight and energy intake in DIO rats and increased the expression of CART peptide in ARC. The plasma ACTH was increased in response to restraint stress, but EA produced no further increase in ACTH levels. The results suggest that EA can up-regulate the expression of CART peptide to approach normal level, resulting in an inhibition of food intake and a reduction of body weight in DIO rats.
Assuntos
Núcleo Arqueado do Hipotálamo/metabolismo , Eletroacupuntura/métodos , Regulação da Expressão Gênica/efeitos da radiação , Proteínas do Tecido Nervoso/metabolismo , Obesidade/terapia , Hormônio Adrenocorticotrópico/sangue , Análise de Variância , Animais , Núcleo Arqueado do Hipotálamo/citologia , Núcleo Arqueado do Hipotálamo/efeitos dos fármacos , Núcleo Arqueado do Hipotálamo/efeitos da radiação , Peso Corporal/efeitos dos fármacos , Peso Corporal/efeitos da radiação , Contagem de Células/métodos , Gorduras na Dieta/efeitos adversos , Relação Dose-Resposta à Radiação , Ensaio de Imunoadsorção Enzimática/métodos , Imuno-Histoquímica/métodos , Masculino , Obesidade/induzido quimicamente , Ratos , Ratos Sprague-Dawley , Restrição Física/métodos , Fatores de TempoRESUMO
Neuropeptide B (NPB) is a recently identified endogenous ligand for the orphan G protein-coupled receptors GPR7 and GPR8. NPB mRNA is expressed in the human, rat, and mouse brain. With the use of an antiserum directed against the rat NPB, immunoreactivity to NPB (irNPB) was detected in several discrete areas of the hypothalamus and midbrain. In the hypothalamus, irNPB cells were present in the medial preoptic area and nucleus, ventromedial preoptic nucleus, retrochiasmatic nucleus, paraventricular hypothalamic nucleus, supraoptic nucleus, accessory neurosecretory nuclei, periventricular hypothalamic nucleus, dorsomedial hypothalamic nucleus, supraoptic retrochiasmatic nucleus, lateral hypothalamic area, posterior hypothalamic area, dorsal hypothalamic area, and zona incerta. A few irNPB perikarya were noted in the arcuate nucleus, whereas a dense network of nerve fibers was present in the median eminence. In the midbrain, irNPB somata were noted in the substantia nigra (compact, reticular, medial, and lateral parts), paranigral nucleus, ventral tegmental area, interfascicular nucleus, and dorsal raphe nucleus. Neurons in the Edinger-Westphal were strongly labeled. Labeled cells were not detected in the cortex, medulla oblongata, and spinal cord; few lightly labeled cells were occasionally seen in the hippocampus. Double labeling the hypothalamic sections with NPB antiserum and vasopressin or oxytocin antibody revealed that a population of vasopressin- but not oxytocin-immunoreactive cells was irNPB. Tyrosine hydroxylase-positive neurons in the midbrain, presumably dopaminergic, were irNPB. The distribution of irNPB neurons in several areas of the hypothalamus and midbrain together with the colocalization with vasopressin or tyrosine hydroxylase suggests that the peptide may subserve neuroendocrine, autonomic, and motor functions.
Assuntos
Sistema Nervoso Central/metabolismo , Neurônios/metabolismo , Neuropeptídeos/biossíntese , Animais , Sistema Nervoso Autônomo/anatomia & histologia , Sistema Nervoso Autônomo/metabolismo , Mapeamento Encefálico , Sistema Nervoso Central/anatomia & histologia , Dopamina/metabolismo , Hipotálamo/anatomia & histologia , Hipotálamo/metabolismo , Imuno-Histoquímica , Masculino , Mesencéfalo/anatomia & histologia , Mesencéfalo/metabolismo , Ratos , Ratos Sprague-Dawley , Receptores de Neuropeptídeos/metabolismo , Tirosina 3-Mono-Oxigenase/metabolismo , Vasopressinas/metabolismoRESUMO
OBJECTIVE: To investigate the effect of electroacupuncture (EA) on rat with diet-induced obesity (DIO) and to explore the possible neurochemical mechanisms using the technique of immumohistochemistry. METHODS: To establish DIO rat model by feeding the animals with high fat diet for 14 weeks. DIO rats were randomly divided into 4 groups: (1) 2Hz EA group, (2) 100Hz EA group, (3) restrain control group, (4) diet resistance (DR) group, (5) DIO group and (6) normal control group. EA treatment: (1) The acupoints used were Zusanli and Sanyinjiao on both legs. (2) The intensities of stimulation were 0.5, 1.0 and 1.5mA for 10 mins each. EA treatment was administered 3 times per week. Food intake and body weight were measured daily for 4 weeks. (3) The changes of the expression of alpha-melanocyte-stimulating hormone (alpha-MSH) in the hypothalamic arcuate nucleus (ARC) were measured with immunohistochemical semiquantitative analysis. RESULTS: (1) The food intake and body weight of 2 Hz EA group and 100 Hz EA group were decreased significantly compared with the restrain control group and DIO group. (2) The number of alpha-MSH positive cells in hypothalamic ARC in 2 Hz EA and 100 Hz EA group was significantly higher than that in restrain control group and DIO group. The number of alpha-MSH positive cells in hypothalamic ARC in DIO group is significantly lower than those in DR group or normal control group. CONCLUSION: A decrease of alpha-MSH level in hypothalamus may be associated with diet-induced obesity. The therapeutic effect on obesity produced by EA may be accounted for by the stimulation of pro-opio-melanocortin neurons in hypothalamic ARC to release alpha-MSH, which inhibits food intake, resulting in a decrease of body weight.
Assuntos
Núcleo Arqueado do Hipotálamo/metabolismo , Eletroacupuntura , Obesidade , alfa-MSH/metabolismo , Animais , Peso Corporal , Dieta , Hipotálamo/metabolismo , Ratos , Regulação para CimaRESUMO
Beacon is a 73-amino acid peptide encoded by a novel gene in the hypothalamus of Israeli sand rat Psammomys obesus. Reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemical techniques were used to investigate the presence of beacon mRNA and the distribution of beacon-immunoreactivity (irBC) in the hypothalamus of ICR mice. RT-PCR experiments revealed beacon mRNA in the mouse hypothalamus. Using a rabbit polyclonal antiserum directed against the synthetic C-terminal peptide fragment (47-73), irBC was detected in the mouse hypothalamus and pituitary. In the hypothalamus, irBC was concentrated in perikarya of the supraoptic (SO), paraventricular (PVH) and accessory neurosecretory nuclei and in cell processes of the median eminence and pituitary stalk. In the pituitary, irBC was noted mainly in the posterior lobe. Double-labeling the hypothalamic sections with guinea-pig vasopressin-antiserum or mouse monoclonal oxytocin-antibody and beacon-antiserum revealed that <30% of vasopressin-immunoreactive neurons and nearly all oxytocin-immunoreactive neurons in the PVH and SO were irBC. The result shows the presence of beacon mRNA in the mouse hypothalamus, and the distribution of irBC is distinctively different from that reported in the hypothalamus of Psammomys obesus, but similar to that of the Sprague-Dawley rats described in our earlier study. More interestingly, Blast search uncovered a 73-amino acid peptide, human ubiquitin-like 5, which has the same exact sequence as beacon. Thus, irBC observed in the mouse brain could be that of ubiquitin-like 5.
Assuntos
Proteínas do Olho/análise , Hipotálamo/química , Proteínas do Tecido Nervoso/análise , Hipófise/química , Ubiquitinas/análise , Animais , Proteínas do Olho/biossíntese , Feminino , Humanos , Hipotálamo/metabolismo , Imuno-Histoquímica , Masculino , Camundongos , Camundongos Endogâmicos ICR , Proteínas do Tecido Nervoso/biossíntese , Hipófise/metabolismo , RNA Mensageiro/análise , RNA Mensageiro/biossíntese , Ubiquitinas/biossínteseRESUMO
Neuropeptide W-23 (NPW23) and neuropeptide W-30 (NPW30) are 23- and 30-amino acid peptides recently isolated from the porcine hypothalamus. Immunohistochemical studies using a rabbit polyclonal antiserum against the rat NPW23 peptide revealed a limited distribution in the rat brain. NPW23-immunoreactive (irNPW) cells were detected in the paraventricular nucleus (PVH), mainly in the parvocellular division, supraoptic nucleus (SO), accessory neurosecretory nuclei, dorsal and lateral hypothalamic areas, perifornical nucleus, arcuate nucleus, and anterior and posterior pituitary; whereas, irNPW fibers were noted in the PVH and SO, retrochiasmatic nucleus, dorsal and lateral hypothalamic areas, median eminence, amygdala, and posterior pituitary. The pattern of distribution of irNPW in the hypothalamus corroborates a possible role of NPW on prolactin release and feeding behavior reported by others.
Assuntos
Hipotálamo/metabolismo , Neuropeptídeos/metabolismo , Hipófise/metabolismo , Animais , Feminino , Imuno-Histoquímica , Masculino , Neurônios/metabolismo , Ratos , Ratos Sprague-DawleyRESUMO
With the use of an antiserum against human apelin-36, apelin-immunoreactivity (irAP) was detected in neurons and cell processes of the supraoptic nucleus (SO), paraventricular nucleus (PVH), accessory neurosecretory nuclei (Acc) and suprachiasmatic nucleus. Strongly labeled cells/processes were noted in the internal layer of the median eminence, infundibular stem, anterior and posterior pituitary. Double-labeling the sections with goat polyclonal neurophysin I-antiserum and rabbit polyclonal apelin-antiserum revealed a population of magnocellular neurons in the PVH, SO and Acc expressing both irAP and neurophysin I-immunoreactivity (irNP), the latter being a marker of oxytocin-containing neurons. By inference, the AP-positive but irNP-negative magnocellular neurons could be vasopressin-containing. The presence of irAP in certain hypothalamic nuclei and pituitary suggests that the peptide may be a signaling molecule released from the hypothalamic-hypophysial axis.
Assuntos
Proteínas de Transporte/análise , Hipotálamo/química , Hipófise/química , Animais , Apelina , Feminino , Imuno-Histoquímica , Peptídeos e Proteínas de Sinalização Intercelular , Masculino , Microscopia Confocal , Neurofisinas/análise , Ratos , Ratos Sprague-DawleyRESUMO
Distribution of the novel peptide beacon in the hypothalamus of Sprague-Dawley rats was examined by immunohistochemical methods. Beacon-immunoreactive (irBC) neurons were found in the paraventricular, supraoptic, and accessory neurosecretory nuclei, and intensely labeled fibers in the median eminence and infundibulo-pituitary stalk. Scattered cells and/or fibers were noted in the suprachiasmatic nucleus, arcuate nucleus, retrochiasmatic area, lateral and medial preoptic area, as well as anterior and lateral hypothalamic area. The wide distribution of irBC in the hypothalamus of Sprague-Dawley rats suggests that the peptide may influence, in addition to a proposed role in feeding, a multitude of biological activities associated with the hypothalamic-pituitary axis.