RESUMO
After fasting, satiety is observed within 2 h after reintroducing food, accompanied by activation of anorexigenic, pro-opiomelanocortin (POMC)-synthesising neurones in the arcuate nucleus (ARC), indicative of the critical role that α-melanocyte-stimulating hormone has in the regulation of meal size during refeeding. To determine whether refeeding-induced activation of POMC neurones in the arcuate is dependent upon the vagus nerve and/or ascending brainstem pathways, bilateral subdiaphragmatic vagotomy or transection of the afferent brainstem input to one side of the ARC was performed. One day after vagotomy or 2 weeks after brain surgery, animals were fasted and then refed for 2 h. Sections containing the ARC from vagotomised animals or animals with effective transection were immunostained for c-Fos and POMC to detect refeeding-induced activation of POMC neurones. Quantitative analyses of double-labelled preparations demonstrated that sham-operated and vagotomised animals markedly increased the number of c-Fos-immunoreactive (-IR) POMC neurones with refeeding. Furthermore, transection of the ascending brainstem pathway had no effect on diminishing c-Fos-immunoreactivity in POMC neurones on either side of the ARC, although it did diminish activation in a separate, subpopulation of neurones in the dorsomedial posterior ARC (dmpARC) on the transected side. We conclude that inputs mediated via the vagus nerve and/or arising from the brainstem do not have a primary role in refeeding-induced activation of POMC neurones in the ARC, and propose that these neurones may be activated solely by direct effects of circulating hormones/metabolites during refeeding. Activation of the dmpARC by refeeding indicates a previously unrecognised role for these neurones in appetite regulation in the rat.
Assuntos
Tronco Encefálico/fisiologia , Ingestão de Alimentos/fisiologia , Neurônios/metabolismo , Neurônios/fisiologia , Pró-Opiomelanocortina/metabolismo , Nervo Vago/fisiologia , Animais , Anorexia/metabolismo , Depressores do Apetite/metabolismo , Regulação do Apetite/fisiologia , Núcleo Arqueado do Hipotálamo/metabolismo , Núcleo Arqueado do Hipotálamo/fisiologia , Tronco Encefálico/metabolismo , Ingestão de Líquidos/fisiologia , Jejum , Masculino , Ratos , Ratos Sprague-Dawley , Transmissão Sináptica/fisiologia , Nervo Vago/metabolismoRESUMO
Infectious diseases and the administration of bacterial lipopolysaccharide (LPS) result in decreased food intake and increased energy expenditure. Because the hypothalamic paraventricular nucleus (PVN) has pivotal roles in the regulation of energy homeostasis and expresses an anorexic peptide, cocaine- and amphetamine-regulated transcript (CART), we hypothesised that increased CART synthesis in this nucleus may contribute to LPS-induced changes in energy homeostasis. Therefore, we studied the effects of intraperitoneal administration of LPS on CART gene expression in the PVN by semiquantitative in situ hybridisation. LPS caused a rapid increase in CART mRNA levels in the PVN. One hour after treatment, the density of silver grains was increased by three-fold in the PVN, and remained elevated 3 h after treatment. Because the dorsal vagal complex, an important vegetative centre in the brainstem, is heavily innervated by CART-containing axons, we determined whether the retrograde tracer, cholera toxin B subunit (CTB), accumulates in CART neurons in the PVN following stereotaxic injection of the tracer into the dorsal vagal complex. One week after injection, CTB accumulated in CART neurons in the ventral, medial, and lateral parvocellular subdivisions of the PVN. In addition, LPS administration induced c-fos expression in a population of CART neurons in the PVN that project to the dorsal vagal complex. These data indicate that increased CART gene expression in neurons of PVN may contribute to LPS-induced anorexia, and suggest that this action may be mediated, at least in part, through a PVN-dorsal vagal complex pathway.
Assuntos
Metabolismo Energético/efeitos dos fármacos , Homeostase/efeitos dos fármacos , Lipopolissacarídeos/farmacologia , Proteínas do Tecido Nervoso/genética , Neurônios/efeitos dos fármacos , Núcleo Hipotalâmico Paraventricular/efeitos dos fármacos , Animais , Anorexia/induzido quimicamente , Anorexia/genética , Endotoxinas/farmacologia , Metabolismo Energético/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Homeostase/genética , Masculino , Proteínas do Tecido Nervoso/metabolismo , Neurônios/metabolismo , Núcleo Hipotalâmico Paraventricular/metabolismo , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Ratos WistarAssuntos
Arte/história , Bócio/história , Bizâncio , Feminino , História Antiga , Humanos , Iodo/deficiência , Itália , Abastecimento de ÁguaRESUMO
Regulation of the hypothalamic-pituitary-thyroid (HPT) axis is dependent upon the secretion of thyrotropin-releasing hormone (TRH), a tripeptide originating in the hypothalamic paraventricular nucleus (PVN). These so-called hypophysiotropic neurons are under feedback inhibition by circulating levels of thyroid hormone, mediated through interactions with the beta2 thyroid hormone receptor (TRbeta2) and competition with the phosphorylated form of cyclic adenosine 5'-monophosphate response element binding protein (CREB) for a multifunctional binding site in the TRH gene. The non-thyroidal illness syndrome, characterized by low circulating thyroid hormone levels yet suppression of TRH gene expression in hypophysiotropic neurons, is due to alteration in the regulatory factors that modulate TRH gene expression to result in central hypothyroidism. These factors include alpha melanocyte-stimulating hormone (alphaMSH) and cocaine- and amphetamine-regulated transcript (CART), and agouti-related protein (AGRP) and neuropeptide Y (NPY), substances co-produced by distinct populations of leptin-responsive neurons in the hypothalamic arcuate nucleus. Through monosynaptic projections from arcuate nucleus neurons to hypophysiotropic TRH neurons, these factors contribute to suppression of HPT axis during fasting and starvation by exerting opposing actions on the TRH gene, altering the sensitivity for feedback inhibition by thyroid hormone. In contrast, central hypothyroidism associated with infection may be due to upregulation of type 2 deiodinase activity in tanycytes, specialized glial cells that line the infralateral walls and floor of the third ventricle. Through tanycyte-cerebrospinal fluid, -vascular or -neuronal associations, these cells may lead to inhibition of TRH gene expression in hypophysiotropic neurons by increasing local triiodothyronine production.
Assuntos
Glândula Tireoide/fisiopatologia , Hormônio Liberador de Tireotropina/fisiologia , Animais , Retroalimentação , Humanos , Hipotálamo/anatomia & histologia , Hipotálamo/fisiopatologia , Infecções/fisiopatologia , Neurônios/fisiologia , Neuro-Hipófise/fisiopatologia , Receptores dos Hormônios Tireóideos/fisiologia , Glândula Tireoide/anatomia & histologia , Hormônio Liberador de Tireotropina/metabolismoRESUMO
Obesity may be an independent risk factor for coronary artery disease and contribute to a chronic state of systemic inflammation leading to atherosclerosis and metabolic abnormalities, such as diabetes, insulin resistance, dyslipidemia and hypertension. Visceral fat, in fact, may act as an endocrine organ, synthesizing and releasing atherogenic inflammatory cytokines, whose circulating levels depend on the individual's nutritional state, and the extent and anatomical location of fat stores. Unsuspected viral infections might also be involved in enhancing autocrine/paracrine mechanisms of cytokine release from omental fat. Elevated levels of blood cytokines may interact with the neuroendocrine system, autonomic nerves and peripheral lymphatic organs. This may lead to local inflammatory reactions in many body compartments, in particular in the heart tissue, possibly affecting the process of circulatory recovery in obese subjects, and predisposing these patients to a greater risk of myocardial inflammatory disease than individuals with normal body mass index. Circulating levels of inflammatory cytokines might be considered to determine risk categories for development of cardiovascular complications in obese subjects. In addition, their reduction with pharmacological antagonists might prevent and/or control acute cardiovascular events and increase energy expenditure in obese patients, especially after surgical treatment, through reduction of cytokine inhibition of the hypothalamic-pituitary-thyroid axis.
Assuntos
Adipócitos/metabolismo , Inflamação/etiologia , Sistemas Neurossecretores/fisiopatologia , Obesidade/complicações , Obesidade/fisiopatologia , Animais , Arteriosclerose/complicações , Arteriosclerose/etiologia , Doença das Coronárias/complicações , Doença das Coronárias/etiologia , Citocinas/efeitos adversos , Citocinas/sangue , Complicações do Diabetes , Diabetes Mellitus/etiologia , Humanos , Hiperlipidemias/complicações , Hiperlipidemias/etiologia , Hipertensão/complicações , Hipertensão/etiologia , Inflamação/complicações , Inflamação/metabolismo , Inflamação/fisiopatologia , Resistência à Insulina , Obesidade/metabolismo , Fatores de RiscoRESUMO
Our previous study has shown that prothyrotropin-releasing hormone (proTRH) gene expression is increased in the ventrolateral periaqueductal gray (PAG) neurons following precipitated morphine withdrawal and continues to be activated even 24 h after withdrawal. We have hypothesized that peptide products of proTRH may participate in the recovery from morphine withdrawal. To identify neuroanatomical substrates of the proposed action of proTRH-derived peptides originating from the ventrolateral PAG proTRH neurons, projections of these neurons were investigated by a series of anterograde and retrograde tract-tracing experiments. First, Phaseolus vulgaris-leucoagglutinin (PHA-L) was injected in the ventrolateral PAG in Sprague-Dawley rats. Following transport of the tracer, simultaneous immunolabeling for PHA-L and proTRH peptides was performed and mapped in discrete brain regions. PHA-L-immunoreactive (IR) fibers showing preterminal and terminal-like arborization that contained proTRH were identified in the dorsolateral and lateral PAG, deep layer of superior colliculus (CS), parafascicular nucleus (PF), ventromedial zona incerta (ZI) and at the border of the locus coeruleus (LC) and Barrington's nucleus. Scattered double-labeled fibers were present in the lateral septal nucleus, ventromedial preoptic nucleus, lateral hypothalamus, perifornical area and in the periventricular region at the diencephalon/midbrain junction. The retrogradely transported marker, cholera toxin beta-subunit (CTb) was then injected in the dorsolateral PAG, CS, PF, ZI and medial to the LC. Double-labeled perikarya for both CTb and proTRH in the ventrolateral PAG were found for each region injected with CTb, corroborating the findings by the anterograde tracing experiment. These studies demonstrate that proTRH neurons in the ventrolateral PAG project to several regions of the brain that are involved in autonomic and behavioral regulation and thereby, may function as an integrating center to coordinate responses to opiate withdrawal.
Assuntos
Vias Eferentes/citologia , Neurônios/citologia , Transtornos Relacionados ao Uso de Opioides/metabolismo , Substância Cinzenta Periaquedutal/citologia , Precursores de Proteínas/metabolismo , Síndrome de Abstinência a Substâncias/metabolismo , Hormônio Liberador de Tireotropina/metabolismo , Animais , Sistema Nervoso Autônomo/citologia , Sistema Nervoso Autônomo/metabolismo , Transporte Axonal/fisiologia , Encéfalo/citologia , Encéfalo/metabolismo , Vias Eferentes/metabolismo , Imunofluorescência , Masculino , Microinjeções , Neurônios/metabolismo , Transtornos Relacionados ao Uso de Opioides/patologia , Transtornos Relacionados ao Uso de Opioides/fisiopatologia , Substância Cinzenta Periaquedutal/metabolismo , Fito-Hemaglutininas , Terminações Pré-Sinápticas/metabolismo , Terminações Pré-Sinápticas/ultraestrutura , Ácido Pirrolidonocarboxílico/análogos & derivados , Ratos , Ratos Sprague-Dawley , Síndrome de Abstinência a Substâncias/patologia , Síndrome de Abstinência a Substâncias/fisiopatologiaRESUMO
Brain-derived neurotrophic factor has been associated previously with the regulation of food intake. To help elucidate the role of this neurotrophin in weight regulation, we have generated conditional mutants in which brain-derived neurotrophic factor has been eliminated from the brain after birth through the use of the cre-loxP recombination system. Brain-derived neurotrophic factor conditional mutants were hyperactive after exposure to stressors and had higher levels of anxiety when evaluated in the light/dark exploration test. They also had mature onset obesity characterized by a dramatic 80-150% increase in body weight, increased linear growth, and elevated serum levels of leptin, insulin, glucose, and cholesterol. In addition, the mutants had an abnormal starvation response and elevated basal levels of POMC, an anorexigenic factor and the precursor for alpha-MSH. Our results demonstrate that brain derived neurotrophic factor has an essential maintenance function in the regulation of anxiety-related behavior and in food intake through central mediators in both the basal and fasted state.
Assuntos
Fator Neurotrófico Derivado do Encéfalo/genética , Encéfalo/metabolismo , Deleção de Genes , Hipercinese/genética , Obesidade/genética , Animais , Ansiedade/genética , Peso Corporal/genética , Jejum , Fluoxetina/farmacologia , Expressão Gênica , Hiperglicemia/genética , Hiperinsulinismo/genética , Hipotálamo/química , Hipotálamo/metabolismo , Integrases/genética , Leptina/análise , Camundongos , Neuropeptídeo Y/genética , Pró-Opiomelanocortina/genética , RNA Mensageiro/análise , Serotonina/metabolismo , Inibidores Seletivos de Recaptação de Serotonina/farmacologia , Transfecção , Proteínas Virais/genéticaRESUMO
To investigate a possible neurotropic role for thyrotropin-releasing hormone (TRH) in the central nervous system, we used recombinant antisense TRH adenovirus (TRHav) to "knock out" TRH in cultured 17-d fetal rat diencephalon. The morphology along with beta-galactosidase (beta-gal) enzyme histochemistry (X-gal staining) and TRH content (femtomoles/well) were used to measure the effect of antisense TRH virus. Control adenovirus mediated beta-gal transfection efficiency was nearly 85%, as shown by positive X-gal staining, and was without effect on cell morphology, TRH content, or the normal response to glucocorticoid (dexamethasone) exposure with enhanced TRH expression. A significant 90% decline in TRH content as well as changes in neuronal morphology (shrunken cell bodies and short dendrites) were observed after 14 but not 7 d following TRHav treatment. The addition of synthetic TRH peptide at 2.5 microM along with TRHav, but not dexamethasone, partly prevented the morphologic changes. No morphologic changes were seen in wild-type AtT20 cells, a pituitary cell line that does not produce TRH. To investigate whether neuronal death from loss of proTRH was owing to apoptosis, neuronal DNA change by means of fluorescent dye H-33342 staining, TUNEL staining, and DNA laddering analysis was examined. Eighty to 90% positive H-33342 and TUNEL staining as well as a 180- to 200-bp DNA fragment on DNA laddering analysis were found as compared to control. These results indicate that proTRH gene expression prevents neuronal apoptosis and may play a role in neuronal development and function.
Assuntos
Sobrevivência Celular , DNA Antissenso/farmacologia , Diencéfalo/citologia , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Precursores de Proteínas/genética , Hormônio Liberador de Tireotropina/genética , Adenoviridae/genética , Animais , Apoptose , Células Cultivadas , Fragmentação do DNA , Embrião de Mamíferos , Hipotálamo/citologia , Marcação In Situ das Extremidades Cortadas , Ratos , Hormônio Liberador de Tireotropina/biossíntese , Transfecção , beta-Galactosidase/genéticaRESUMO
The goal of the present investigation was to analyze the types 2 (D2) and 3 (D3) iodothyronine deiodinases in various structures within the central nervous system (CNS) in response to iodine deficiency. After 5-6 wk of low-iodine diet (LID) or LID + 2 microg potassium iodide/ml (LID + KI; control), rats' brains were processed for in situ hybridization histochemistry for D2 and D3 mRNA or dissected, frozen in liquid nitrogen, and processed for D2 and D3 activities. LID did not affect weight gain or serum triiodothyronine, but plasma thyroxine (T4) was undetectable. In the LID + KI animals, D3 activities were highest in the cerebral cortex (CO) and hippocampus (HI), followed by the olfactory bulb and was lowest in cerebellum (CE). Iodine deficiency decreased D3 mRNA expression in all CNS regions, and these changes were accompanied by three- to eightfold decreases in D3 activity. In control animals, D2 activity in the medial basal hypothalamus (MBH) was similar to that in pituitary gland. Of the CNS D2-expressing regions analyzed, the two most responsive to iodine deficiency were the CO and HI, in which an approximately 20-fold increase in D2 activity occurred. Other regions, i.e., CE, lateral hypothalamus, MBH, and pituitary gland, showed smaller increases. The distribution of and changes in D2 mRNA were similar to those of D2 activity. Our results indicate that decreases in the expression of D3 and increases in D2 are an integral peripheral component of the physiological response of the CNS to iodine deficiency.
Assuntos
Adaptação Fisiológica/fisiologia , Sistema Nervoso Central/fisiologia , Iodeto Peroxidase/metabolismo , Iodo/deficiência , Animais , Dieta , Hibridização In Situ , Iodeto Peroxidase/biossíntese , Isoenzimas/metabolismo , Masculino , Tamanho do Órgão/fisiologia , RNA Mensageiro/biossíntese , Ratos , Ratos Sprague-Dawley , Glândula Tireoide/patologia , Hormônios Tireóideos/sangueRESUMO
Recent evidence suggests that neuropeptide Y (NPY), originating in neurons in the hypothalamic arcuate nucleus, is an important mediator of the effects of leptin on the central nervous system. As these NPY neurons innervate hypophysiotropic neurons in the hypothalamic paraventricular nucleus (PVN) that produce the tripeptide, TRH, we raised the possibility that NPY may be responsible for resetting of the hypothalamic-pituitary-thyroid (HPT) axis during fasting. To test this hypothesis, the effects of intracerebroventricularly administered NPY on circulating thyroid hormone levels and proTRH messenger RNA in the PVN were studied by RIA and in situ hybridization histochemistry, respectively. NPY administration suppressed circulating levels of thyroid hormone (T(3) and T(4)) and resulted in an inappropriately normal or low TSH. These alterations were associated with a significant suppression of proTRH messenger RNA in the PVN, indicating that NPY infusion had resulted in a state of central hypothyroidism. Similar observations were made in NPY-infused animals pair fed to the vehicle-treated controls. These data are reminiscent of the effect of fasting on the thyroid axis and indicate that NPY may play a major role in the inhibition of HPT axis during fasting.
Assuntos
Hipotálamo/efeitos dos fármacos , Neuropeptídeo Y/farmacologia , Hipófise/efeitos dos fármacos , Glândula Tireoide/efeitos dos fármacos , Tecido Adiposo/efeitos dos fármacos , Tecido Adiposo Marrom/efeitos dos fármacos , Animais , Núcleo Arqueado do Hipotálamo/química , Autorradiografia , Peso Corporal/efeitos dos fármacos , Encéfalo/efeitos dos fármacos , Ingestão de Alimentos/efeitos dos fármacos , Epididimo , Hipotálamo/fisiologia , Hibridização In Situ , Leptina/análise , Masculino , Neuropeptídeo Y/administração & dosagem , Tamanho do Órgão/efeitos dos fármacos , Hipófise/fisiologia , Precursores de Proteínas/genética , RNA Mensageiro/análise , Ratos , Ratos Sprague-Dawley , Glândula Tireoide/fisiologia , Tireotropina/sangue , Tireotropina/genética , Tiroxina/sangue , Tri-Iodotironina/sangueRESUMO
To determine whether the hypothalamic dorsomedial nucleus (DMN) may serve as a relay center for the central actions of leptin on thyrotropin-releasing hormone (TRH)-synthesizing neurons in the paraventricular nucleus (PVN), axonal projections from the DMN to TRH-containing neurons in the PVN were studied using the anterogradely transported marker substance, Phaseolus vulgaris-leucoagglutinin (PHA-L). Stereotaxic injections of PHA-L were targeted to the mid-dorsal and mid-ventral portions of the DMN. After 10-14-day survival, the brains were prepared for immunohistochemistry and immunostained with an antibody directed against PHA-L. Focal injections confined to the DMN were identified in 14 animals and gave rise to a fiber bundle that entered the PVN at the caudal pole of the nucleus, densely innervating all parvocellular subdivisions of the PVN. In double-labeled preparations using antisera to PHA-L and preproTRH 178-199, the latter as a marker for TRH-containing neurons in the PVN, proTRH-IR neurons were observed to be enmeshed in a network of PHA-L-containing fibers. When the injection site covered the entire DMN or the mid-dorsal part of the DMN, PHA-L-containing axon varicosities were juxtaposed to approximately 97 and 90% of proTRH neurons, respectively, in all parvocellular subdivisions of the PVN, and by ultrastructural analysis were shown to be synaptic. In contrast, when the injection site was centered primarily in the mid-ventral part of the DMN, only approximately 52% of proTRH-synthesizing neurons appeared to be innervated by PHA-L-containing axons. These data demonstrate that a major projection pathway exists from the DMN, specifically to TRH-producing neurons in the PVN, and suggest that the DMN is anatomically situated to exert a regulatory effect on TRH-synthesizing neurons in the PVN.
Assuntos
Núcleo Hipotalâmico Dorsomedial/ultraestrutura , Vias Neurais/ultraestrutura , Neurônios/ultraestrutura , Núcleo Hipotalâmico Paraventricular/ultraestrutura , Hormônio Liberador de Tireotropina/metabolismo , Animais , Núcleo Hipotalâmico Dorsomedial/metabolismo , Imuno-Histoquímica , Leptina/metabolismo , Masculino , Microscopia Eletrônica , Vias Neurais/metabolismo , Neurônios/citologia , Núcleo Hipotalâmico Paraventricular/metabolismo , Fito-Hemaglutininas/farmacologia , Terminações Pré-Sinápticas/metabolismo , Terminações Pré-Sinápticas/ultraestrutura , Ratos , Ratos Sprague-Dawley , Sinapses/metabolismo , Sinapses/ultraestrutura , alfa-MSH/metabolismoRESUMO
We recently demonstrated that three arcuate nucleus-derived peptides, neuropeptide Y (NPY), agouti-related protein (AGRP), and alphaMSH, are contained in axon terminals that heavily innervate hypophysiotropic TRH neurons in the rat brain and may contribute to the altered set-point of the hypothalamo-pituitary-thyroid axis during fasting. To determine whether a similar regulatory system exists in human brain, we performed a series of immunohistochemical studies using antisera against NPY, AGRP, alphaMSH, and TRH in adult hypothalami obtained within 15 h of death. Numerous small to medium-sized, fusiform and multipolar NPY-, AGRP-, and alphaMSH-immunoreactive (-IR) cells were widely distributed throughout the rostro-caudal extent of the infundibular (arcuate) nucleus. A similar distribution pattern was found for NPY- and AGRP-IR neurons in the arcuate nucleus, whereas alphaMSH-IR cells appeared to form a separate cell population. By double labeling fluorescent immunohistochemistry, 82% of NPY neurons cocontained AGRP, and 87% of AGRP neurons coexpressed NPY. No colocalization was found between alphaMSH- and AGRP-IR neurons. NPY-, AGRP-, and alphaMSH-containing axons densely innervated the hypothalamic paraventricular nucleus and were found in close juxtaposition to TRH-synthesizing cell bodies and dendrites. These studies demonstrate that in man, the NPY-, AGRP-, and alphaMSH-IR neuronal systems in the infundibular and paraventricular nuclei are highly reminiscent of that observed in the rat and may similarly be involved in regulating the hypothalamo-pituitary-thyroid axis in the human brain.
Assuntos
Hipotálamo/metabolismo , Neurônios/metabolismo , Neuropeptídeo Y/fisiologia , Proteínas/fisiologia , Hormônio Liberador de Tireotropina/biossíntese , alfa-MSH/fisiologia , Adulto , Proteína Relacionada com Agouti , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Hipotálamo/citologia , Imuno-Histoquímica , Peptídeos e Proteínas de Sinalização Intercelular , Masculino , Pessoa de Meia-Idade , Fibras Nervosas/metabolismo , Núcleo Hipotalâmico Paraventricular/citologia , Núcleo Hipotalâmico Paraventricular/metabolismo , Núcleo Hipotalâmico Paraventricular/fisiologiaRESUMO
During fasting, corticotropin-releasing hormone (CRH) mRNA decreases in the hypothalamic paraventricular nucleus (PVN), but the mechanism by which this takes place is not well understood. To test the hypothesis that the melanocortin system may be involved in the regulation of CRH mRNA in the PVN during fasting, the effect of intracerebroventricularly administered alpha-melanocyte stimulating hormone (MSH) on CRH mRNA in the PVN was studied in fasted animals by in situ hybridization histochemistry. Whereas fasting suppressed CRH mRNA levels in the PVN, alpha-MSH at doses of 150 and 300 ng every 6 h for 64 h prevented the fasting-induced suppression of CRH gene expression in the PVN. These data indicate that the suppression of alpha-MSH synthesis may be responsible for the decreased CRH gene expression in the PVN during fasting.
Assuntos
Hormônio Liberador da Corticotropina/antagonistas & inibidores , Hormônio Liberador da Corticotropina/genética , Jejum/fisiologia , Regulação da Expressão Gênica/fisiologia , Núcleo Hipotalâmico Paraventricular/metabolismo , alfa-MSH/fisiologia , Animais , Hormônio Liberador da Corticotropina/fisiologia , Hibridização In Situ , Masculino , RNA Mensageiro/análise , Ratos , Ratos Sprague-DawleyRESUMO
Type 2 iodothyronine deiodinase, an enzyme involved in the conversion of thyroxin to the biologically active 3,5, 3'-triiodothyronine, is highly concentrated in a group of specialized ependymal cells, tanycytes, lining the wall and floor of the third ventricle. As this distribution is highly reminiscent of the distribution of cells containing the phosphatase inhibitor, DARPP-32, we raised the possibility that these two proteins may coexist in tanycytes and that DARPP-32 may modulate type 2 deiodinase activity by regulating the phosphorylation state of the cAMP regulatory factor, CREB. To address this question, double-labeling histochemical studies were performed for type 2 deiodinase mRNA and DARPP-32 immunoreactivity (IR), or DARPP-32- and CREB-IR in the same tissue sections. Type 2 deiodinase mRNA was found in the cell bodies of all DARPP-32-immunolabeled tanycytes. Both type 2 deiodinase mRNA and DARPP-32-IR also extended into tanycyte processes that ramified in the arcuate nucleus and median eminence, in close association with blood vessels and portal capillaries. In contrast, type 2 deiodinase mRNA was not present in the same cells that contained DARPP-32-IR in the pituitary gland. All tanycytes containing DARPP-32-IR also contained CREB-IR in their nucleus. Since type 2 deiodinase activity can be induced by substances that increase cAMP, we hypothesize that DARPP-32 may regulate the activity of type 2 deiodinase by prolonging the activation of CREB. Selectivity for the colocalization of these factors to tanycytes but not the pituitary gland, may explain the heterogeneous response of type 2 deiodinase activity in these two loci in response to specific stimuli such as fasting.
Assuntos
Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/análise , Epêndima/química , Epêndima/enzimologia , Iodeto Peroxidase/genética , Fosfoproteínas/análise , Animais , Fosfoproteína 32 Regulada por cAMP e Dopamina , Ativação Enzimática/genética , Epêndima/citologia , Regulação Enzimológica da Expressão Gênica , Imuno-Histoquímica , Hibridização In Situ , Iodeto Peroxidase/análise , Masculino , Proteínas do Tecido Nervoso/análise , RNA Mensageiro/análise , Ratos , Ratos Sprague-Dawley , Radioisótopos de Enxofre , Iodotironina Desiodinase Tipo IIRESUMO
The hypothalamic arcuate nucleus has an essential role in mediating the homeostatic responses of the thyroid axis to fasting by altering the sensitivity of prothyrotropin-releasing hormone (pro-TRH) gene expression in the paraventricular nucleus (PVN) to feedback regulation by thyroid hormone. Because agouti-related protein (AGRP), a leptin-regulated, arcuate nucleus-derived peptide with alpha-MSH antagonist activity, is contained in axon terminals that terminate on TRH neurons in the PVN, we raised the possibility that alpha-MSH may also participate in the mechanism by which leptin influences pro-TRH gene expression. By double-labeling immunocytochemistry, alpha-MSH-IR axon varicosities were juxtaposed to approximately 70% of pro-TRH neurons in the anterior and periventricular parvocellular subdivisions of the PVN and to 34% of pro-TRH neurons in the medial parvocellular subdivision, establishing synaptic contacts both on the cell soma and dendrites. All pro-TRH neurons receiving contacts by alpha-MSH-containing fibers also were innervated by axons containing AGRP. The intracerebroventricular infusion of 300 ng of alpha-MSH every 6 hr for 3 d prevented fasting-induced suppression of pro-TRH in the PVN but had no effect on AGRP mRNA in the arcuate nucleus. alpha-MSH also increased circulating levels of free thyroxine (T4) 2.5-fold over the levels in fasted controls, but free T4 did not reach the levels in fed controls. These data suggest that alpha-MSH has an important role in the activation of pro-TRH gene expression in hypophysiotropic neurons via either a mono- and/or multisynaptic pathway to the PVN, but factors in addition to alpha-MSH also contribute to the mechanism by which leptin administration restores thyroid hormone levels to normal in fasted animals.
Assuntos
Jejum/fisiologia , Regulação da Expressão Gênica/fisiologia , Terminações Nervosas/fisiologia , Neurônios/fisiologia , Núcleo Hipotalâmico Paraventricular/fisiologia , Precursores de Proteínas/genética , Hormônio Liberador de Tireotropina/genética , alfa-MSH/fisiologia , Animais , Peso Corporal/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Imuno-Histoquímica , Hibridização In Situ , Masculino , Microscopia Imunoeletrônica , Terminações Nervosas/ultraestrutura , Neurônios/citologia , Neurônios/ultraestrutura , Núcleo Hipotalâmico Paraventricular/citologia , Núcleo Hipotalâmico Paraventricular/ultraestrutura , Precursores de Proteínas/análise , Ácido Pirrolidonocarboxílico/análogos & derivados , Ratos , Ratos Sprague-Dawley , Tireotropina/sangue , Hormônio Liberador de Tireotropina/análise , Tiroxina/sangue , alfa-MSH/análise , alfa-MSH/farmacologiaRESUMO
Because cocaine- and amphetamine-regulated transcript (CART) coexists with alpha-melanocyte stimulating hormone (alpha-MSH) in the arcuate nucleus neurons and we have recently demonstrated that alpha-MSH innervates TRH-synthesizing neurons in the hypothalamic paraventricular nucleus (PVN), we raised the possibility that CART may also be contained in fibers that innervate hypophysiotropic thyrotropin-releasing hormone (TRH) neurons and modulate TRH gene expression. Triple-labeling fluorescent in situ hybridization and immunofluorescence were performed to reveal the morphological relationships between pro-TRH mRNA-containing neurons and CART- and alpha-MSH-immunoreactive (IR) axons. CART-IR axons densely innervated the majority of pro-TRH mRNA-containing neurons in all parvocellular subdivisions of the PVN and established asymmetric synaptic specializations with pro-TRH neurons. However, whereas all alpha-MSH-IR axons in the PVN contained CART-IR, only a portion of CART-IR axons in contact with pro-TRH neurons were immunoreactive for alpha-MSH. In the medial and periventricular parvocellular subdivisions of the PVN, CART was co-contained in approximately 80% of pro-TRH neuronal perikarya, whereas colocalization with pro-TRH was found in <10% of the anterior parvocellular subdivision neurons. In addition, >80% of TRH/CART neurons in the periventricular and medial parvocellular subdivisions accumulated Fluoro-Gold after systemic administration, suggesting that CART may serve as a marker for hypophysiotropic TRH neurons. CART prevented fasting-induced suppression of pro-TRH in the PVN when administered intracerebroventricularly and increased the content of TRH in hypothalamic cell cultures. These studies establish an anatomical association between CART and pro-TRH-producing neurons in the PVN and demonstrate that CART has a stimulatory effect on hypophysiotropic TRH neurons by increasing pro-TRH gene expression and the biosynthesis of TRH.
Assuntos
Jejum/metabolismo , Hipotálamo/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Neurônios/metabolismo , Precursores de Proteínas/biossíntese , Estilbamidinas , Hormônio Liberador de Tireotropina/biossíntese , Animais , Comportamento Animal/efeitos dos fármacos , Peso Corporal/efeitos dos fármacos , Contagem de Células , Células Cultivadas , Colchicina/administração & dosagem , Corantes Fluorescentes , Sistema Hipotálamo-Hipofisário/metabolismo , Hipotálamo/citologia , Hipotálamo/efeitos dos fármacos , Injeções Intraventriculares , Masculino , Proteínas do Tecido Nervoso/administração & dosagem , Neurônios/efeitos dos fármacos , Neurônios/ultraestrutura , Núcleo Hipotalâmico Paraventricular/diagnóstico por imagem , Núcleo Hipotalâmico Paraventricular/efeitos dos fármacos , Núcleo Hipotalâmico Paraventricular/metabolismo , Núcleo Hipotalâmico Paraventricular/ultraestrutura , Terminações Pré-Sinápticas/metabolismo , Terminações Pré-Sinápticas/ultraestrutura , Precursores de Proteínas/genética , Ácido Pirrolidonocarboxílico/análogos & derivados , RNA Mensageiro/metabolismo , Radiografia , Ratos , Ratos Sprague-Dawley , Glândula Tireoide/metabolismo , Tireotropina/sangue , Hormônio Liberador de Tireotropina/genética , Tiroxina/sangue , alfa-MSH/metabolismoRESUMO
Gene expression for agouti-related protein (AGRP), an endogenous antagonist of melanocortin receptors, has been localized to the hypothalamic arcuate nucleus, where it colocalizes with neuropeptide Y (NPY). Having reported that the NPY innervation of hypophysiotropic TRH neurons in the hypothalamic paraventricular nucleus (PVN) originates primarily from NPY-producing neurons in the arcuate nucleus, here we examined the possibility that TRH neurons in the PVN are similarly innervated by AGRP nerve terminals. Using immunohistochemistry, AGRP-containing cell bodies were found almost exclusively in the arcuate nucleus, but their projections were distributed widely in the hypothalamus, most conspicuously in the paraventricular (PVN), arcuate and dorsomedial nuclei, and the posterior hypothalamic area. Ablation of the arcuate nucleus by the neonatal administration of monosodium glutamate obliterated nearly all AGRP-immunoreactivity in the hypothalamus. In the PVN, double-labeling light and electron microscopic immunohistochemistry revealed that TRH neurons receive dense innervation by AGRP nerve terminals, with the frequent occurrence of axosomatic and axodendritic synapses (mainly of the symmetrical type). These findings provide morphological basis to hypothesize a role for AGRP in the arcuato-paraventricular pathway, in the down-regulation of the hypothalamic-pituitary-thyroid axis, which occurs as an adaptive response to starvation.
Assuntos
Terminações Nervosas/fisiologia , Fibras Nervosas/fisiologia , Neurônios/fisiologia , Núcleo Hipotalâmico Paraventricular/fisiologia , Proteínas/fisiologia , Hormônio Liberador de Tireotropina/fisiologia , Proteína Relacionada com Agouti , Animais , Núcleo Arqueado do Hipotálamo/citologia , Núcleo Arqueado do Hipotálamo/efeitos dos fármacos , Núcleo Arqueado do Hipotálamo/fisiologia , Imuno-Histoquímica , Peptídeos e Proteínas de Sinalização Intercelular , Masculino , Microscopia Imunoeletrônica , Terminações Nervosas/ultraestrutura , Fibras Nervosas/ultraestrutura , Neurônios/citologia , Neurônios/efeitos dos fármacos , Núcleo Hipotalâmico Paraventricular/citologia , Núcleo Hipotalâmico Paraventricular/efeitos dos fármacos , Proteínas/análise , Ratos , Ratos Sprague-Dawley , Glutamato de Sódio/farmacologia , Hormônio Liberador de Tireotropina/análiseRESUMO
Type 3 iodothyronine deiodinase (D3) is a selenoenzyme that inactivates thyroid hormone. It is necessary for T3 homeostasis in the central nervous system. D3 activity has been identified in many regions of the brain and parallels thyroid status, but the level at which it is regulated and its specific cellular locations are not known. We evaluated the effect of thyroid status on the expression of the D3 gene within the central nervous system using in situ hybridization histochemistry. D3 messenger RNA (mRNA) was identified throughout, but with high focal expression in the hippocampal pyramidal neurons, granule cells of the dentate nucleus, and layers II-VI of the cerebral cortex. In every region, D3 mRNA abundance was correlated with thyroid status. Four different D3 transcripts were identified by Northern analyses, with evidence for region-specific processing, and D3 mRNA increased 4- to 50-fold from the euthyroid to the hyperthyroid state. D3 mRNA was not detectable in hypothyroid brain. In the central nervous system, the D3 gene is highly T3 responsive, and its focal localization within the hippocampus and cerebral cortex suggests an important role for T3 homeostasis in memory and cognitive functions.