RESUMO
Tanycyte is a subtype of ependymal cells which extend long radial processes to brain parenchyma. The present study showed that tanycyte-like ependymal cells in the organum vasculosum of the lamina terminalis, subfornical organ and central canal (CC) expressed neural stem cell (NSC) marker nestin, glial fibrillar acidic protein and sex determining region Y. Proliferation of these tanycyte-like ependymal cells was promoted by continuous intracerebroventricular infusion of fibroblast growth factor-2 and epidermal growth factor. Tanycytes-like ependymal cells in the CC are able to form self-renewing neurospheres and give rise mostly to new astrocytes and oligodendrocytes. Collagenase-induced small medullary hemorrhage increased proliferation of tanycyte-like ependymal cells in the CC. These results demonstrate that these tanycyte-like ependymal cells of the adult mouse brain are NSCs and suggest that they serve as a source for providing new neuronal lineage cells upon brain damage in the medulla oblongata.
Assuntos
Órgãos Circunventriculares/metabolismo , Células Ependimogliais/metabolismo , Células-Tronco Neurais/metabolismo , Neurônios/metabolismo , Animais , Encéfalo/crescimento & desenvolvimento , Encéfalo/metabolismo , Linhagem da Célula/genética , Proliferação de Células/genética , Órgãos Circunventriculares/crescimento & desenvolvimento , Epêndima/crescimento & desenvolvimento , Epêndima/metabolismo , Células Ependimogliais/citologia , Fator de Crescimento Epidérmico/genética , Fator 2 de Crescimento de Fibroblastos/genética , Regulação da Expressão Gênica/genética , Humanos , Hipotálamo/crescimento & desenvolvimento , Hipotálamo/metabolismo , Camundongos , Nestina/genética , Células-Tronco Neurais/citologia , Organum Vasculosum/crescimento & desenvolvimento , Organum Vasculosum/metabolismo , Órgão Subfornical/crescimento & desenvolvimento , Órgão Subfornical/metabolismoRESUMO
We previously showed that 2 weeks of a severe food restricted (sFR) diet (40% of the caloric intake of the control (CT) diet) up-regulated the circulating renin angiotensin (Ang) system (RAS) in female Fischer rats, most likely as a result of the fall in plasma volume. In this study, we investigated the role of the central RAS in the mean arterial pressure (MAP) and heart rate (HR) dysregulation associated with sFR. Although sFR reduced basal mean MAP and HR, the magnitude of the pressor response to intracerebroventricular (icv) microinjection of Ang-[1-8] was not affected; however, HR was 57 ± 13 bpm lower 26 min after Ang-[1-8] microinjection in the sFR rats and a similar response was observed after losartan was microinjected. The major catabolic pathway of Ang-[1-8] in the hypothalamus was via Ang-[1-7]; however, no differences were detected in the rate of Ang-[1-8] synthesis or degradation between CT and sFR animals. While sFR had no effect on the AT1 R binding in the subfornical organ (SFO), the organum vasculosum laminae terminalis (OVLT) and median preoptic nucleus (MnPO) of the paraventricular anteroventral third ventricle, ligand binding increased 1.4-fold in the paraventricular nucleus (PVN) of the hypothalamus. These findings suggest that sFR stimulates the central RAS by increasing AT1 R expression in the PVN as a compensatory response to the reduction in basal MAP and HR. These findings have implications for people experiencing a period of sFR since an activated central RAS could increase their risk of disorders involving over activation of the RAS including renal and cardiovascular diseases.
Assuntos
Angiotensina I/metabolismo , Pressão Arterial/fisiologia , Restrição Calórica , Frequência Cardíaca/fisiologia , Hipotálamo/metabolismo , Fragmentos de Peptídeos/metabolismo , Receptor Tipo 1 de Angiotensina/metabolismo , Sistema Renina-Angiotensina/fisiologia , Inanição/metabolismo , Angiotensina II/farmacologia , Bloqueadores do Receptor Tipo 1 de Angiotensina II/farmacologia , Enzima de Conversão de Angiotensina 2/metabolismo , Animais , Pressão Arterial/efeitos dos fármacos , Autorradiografia , Feminino , Frequência Cardíaca/efeitos dos fármacos , Injeções Intraventriculares , Losartan/farmacologia , Organum Vasculosum/metabolismo , Núcleo Hipotalâmico Paraventricular/metabolismo , Fragmentos de Peptídeos/farmacologia , Peptidil Dipeptidase A/metabolismo , Área Pré-Óptica/metabolismo , Ratos , Ratos Endogâmicos F344 , Sistema Renina-Angiotensina/efeitos dos fármacos , Órgão Subfornical/metabolismoRESUMO
Angiotensin II (AngII) elicits the production of superoxide (O2â¢-) from mitochondria in numerous cell types within peripheral organs and in the brain suggesting a role for mitochondrial-produced O2â¢- in the pathogenesis of hypertension. However, it remains unclear if mitochondrial O2â¢- is causal in the development of AngII-induced hypertension, or if mitochondrial O2â¢- in the absence of elevated AngII is sufficient to increase blood pressure. Further, the tissue specific (i.e. central versus peripheral) redox regulation of AngII hypertension remains elusive. Herein, we hypothesized that increased mitochondrial O2â¢- in the absence of pro-hypertensive stimuli, such as AngII, elevates baseline systemic mean arterial pressure (MAP), and that AngII-mediated hypertension is exacerbated in animals with increased mitochondrial O2â¢- levels. To address this hypothesis, we generated novel inducible knock-down mouse models of manganese superoxide dismutase (MnSOD), the O2â¢- scavenging antioxidant enzyme specifically localized to mitochondria, targeted to either the brain subfornical organ (SFO) or peripheral tissues. Contrary to our hypothesis, knock-down of MnSOD either in the SFO or in peripheral tissues was not sufficient to alter baseline systemic MAP. Interestingly, when mice were challenged with chronic, peripheral infusion of AngII, only the MnSOD knock-down confined to the SFO, and not the periphery, demonstrated an increased sensitization and potentiated hypertension. In complementary experiments, over-expressing MnSOD in the SFO significantly decreased blood pressure in response to chronic AngII. Overall, these studies indicate that mitochondrial O2â¢- in the brain SFO works in concert with other AngII-dependent factors to drive an increase in MAP, as elevated mitochondrial O2â¢- alone, either in the SFO or peripheral tissues, failed to raise baseline blood pressure.
Assuntos
Angiotensina II/metabolismo , Hipertensão/genética , Superóxido Dismutase/genética , Superóxidos/metabolismo , Angiotensina II/genética , Animais , Antioxidantes/metabolismo , Pressão Sanguínea , Encéfalo/metabolismo , Encéfalo/fisiopatologia , Regulação da Expressão Gênica , Técnicas de Silenciamento de Genes , Humanos , Hipertensão/metabolismo , Hipertensão/fisiopatologia , Camundongos , Mitocôndrias/metabolismo , Mitocôndrias/patologia , Especificidade de Órgãos , Oxirredução , Órgão Subfornical/metabolismo , Órgão Subfornical/patologiaRESUMO
Body fluid conditions are continuously monitored in the brain to regulate thirst and salt-appetite sensations. Angiotensin II drives both thirst and salt appetite; however, the neural mechanisms underlying selective water- and/or salt-intake behaviors remain unknown. Using optogenetics, we show that thirst and salt appetite are driven by distinct groups of angiotensin II receptor type 1a-positive excitatory neurons in the subfornical organ. Neurons projecting to the organum vasculosum lamina terminalis control water intake, while those projecting to the ventral part of the bed nucleus of the stria terminalis control salt intake. Thirst-driving neurons are suppressed under sodium-depleted conditions through cholecystokinin-mediated activation of GABAergic neurons. In contrast, the salt appetite-driving neurons were suppressed under dehydrated conditions through activation of another population of GABAergic neurons by Nax signals. These distinct mechanisms in the subfornical organ may underlie the selective intakes of water and/or salt and may contribute to body fluid homeostasis.
Assuntos
Apetite , Ingestão de Líquidos/fisiologia , Hipotálamo/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Sensação/efeitos dos fármacos , Cloreto de Sódio/farmacologia , Sede/fisiologia , Animais , Apetite/efeitos dos fármacos , Encéfalo/efeitos dos fármacos , Ingestão de Líquidos/efeitos dos fármacos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Neurônios/fisiologia , Órgão Subfornical/metabolismoRESUMO
Water deprivation (WD) induces changes in plasma volume and osmolality, which in turn activate several responses, including thirst, the activation of the renin-angiotensin system (RAS) and vasopressin (AVP) and oxytocin (OT) secretion. These systems seem to be influenced by oestradiol, as evidenced by the expression of its receptor in brain areas that control fluid balance. Thus, we investigated the effects of oestradiol treatment on behavioural and neuroendocrine changes of ovariectomized rats in response to WD. We observed that in response to WD, oestradiol treatment attenuated water intake, plasma osmolality and haematocrit but did not change urinary volume or osmolality. Moreover, oestradiol potentiated WD-induced AVP secretion, but did not alter the plasma OT or angiotensin II (Ang II) concentrations. Immunohistochemical data showed that oestradiol potentiated vasopressinergic neuronal activation in the lateral magnocellular PVN (PaLM) and supraoptic (SON) nuclei but did not induce further changes in Fos expression in the median preoptic nucleus (MnPO) or subfornical organ (SFO) or in oxytocinergic neuronal activation in the SON and PVN of WD rats. Regarding mRNA expression, oestradiol increased OT mRNA expression in the SON and PVN under basal conditions and after WD, but did not induce additional changes in the mRNA expression for AVP in the SON or PVN. It also did not affect the mRNA expression of RAS components in the PVN. In conclusion, our results show that oestradiol acts mainly on the vasopressinergic system in response to WD, potentiating vasopressinergic neuronal activation and AVP secretion without altering AVP mRNA expression.
Assuntos
Desidratação/fisiopatologia , Estradiol/uso terapêutico , Estrogênios/uso terapêutico , Neurônios/efeitos dos fármacos , Núcleo Hipotalâmico Paraventricular/efeitos dos fármacos , Núcleo Supraóptico/efeitos dos fármacos , Desequilíbrio Hidroeletrolítico/prevenção & controle , Animais , Arginina Vasopressina/agonistas , Arginina Vasopressina/análise , Arginina Vasopressina/metabolismo , Comportamento Animal/efeitos dos fármacos , Desidratação/terapia , Ingestão de Líquidos/efeitos dos fármacos , Terapia de Reposição de Estrogênios , Feminino , Hidratação , Regulação da Expressão Gênica/efeitos dos fármacos , Proteínas do Tecido Nervoso/genética , Proteínas do Tecido Nervoso/metabolismo , Neurônios/metabolismo , Neurônios/patologia , Ovariectomia/efeitos adversos , Núcleo Hipotalâmico Paraventricular/metabolismo , Núcleo Hipotalâmico Paraventricular/patologia , Área Pré-Óptica/efeitos dos fármacos , Área Pré-Óptica/metabolismo , Área Pré-Óptica/patologia , Ratos Wistar , Órgão Subfornical/efeitos dos fármacos , Órgão Subfornical/metabolismo , Órgão Subfornical/patologia , Núcleo Supraóptico/metabolismo , Núcleo Supraóptico/patologia , Núcleo Vestibular Lateral/efeitos dos fármacos , Núcleo Vestibular Lateral/metabolismo , Núcleo Vestibular Lateral/patologia , Desequilíbrio Hidroeletrolítico/sangue , Desequilíbrio Hidroeletrolítico/etiologia , Desequilíbrio Hidroeletrolítico/fisiopatologiaRESUMO
Activation of angiotensinergic pathways by central aldosterone (Aldo)-mineralocorticoid receptor (MR) pathway plays a critical role in angiotensin II (Ang II)-induced hypertension. The subfornical organ (SFO) contains both MR and angiotensin II type 1 receptors (AT1R) and can relay the signals of circulating Ang II to downstream nuclei such as the paraventricular nucleus (PVN), supraoptic nucleus (SON) and rostral ventrolateral medulla (RVLM). In Wistar rats, subcutaneous (sc) infusion of Ang II at 500ng/min/kg for 1 or 2weeks increased reactive oxygen species (ROS) as measured by dihydroethidium (DHE) staining in a nucleus - specific pattern. Intra-SFO infusion of AAV-MR- or AT1aR-siRNA prevented the Ang II-induced increase in AT1R mRNA expression in the SFO and decreased MR mRNA. Both MR- and AT1aR-siRNA prevented increases in ROS in the PVN and RVLM. MR- but not AT1aR-siRNA in the SFO prevented the Ang II-induced ROS in the SON. Both MR- and AT1aR-siRNA in the SFO prevented most of the Ang II-induced hypertension as assessed by telemetry. These results indicate that Aldo-MR signaling in the SFO is needed for the activation of Ang II-AT1R-ROS signaling from the SFO to the PVN and RVLM. Activation of Aldo-MR signaling from the SFO to the SON may enhance AT1R dependent activation of pre-sympathetic neurons in the PVN.
Assuntos
Angiotensina II/metabolismo , Hipotálamo/metabolismo , Receptor Tipo 1 de Angiotensina/metabolismo , Receptores de Mineralocorticoides/metabolismo , Órgão Subfornical/metabolismo , Aldosterona/metabolismo , Angiotensina II/administração & dosagem , Animais , Pressão Sanguínea/fisiologia , Dependovirus , Técnicas de Silenciamento de Genes , Vetores Genéticos , Frequência Cardíaca/fisiologia , Hipertensão/metabolismo , Masculino , Bulbo/metabolismo , RNA Mensageiro/metabolismo , RNA Interferente Pequeno , Distribuição Aleatória , Ratos Wistar , Espécies Reativas de Oxigênio/metabolismo , Receptor Tipo 1 de Angiotensina/genética , Receptores de Mineralocorticoides/genéticaRESUMO
In the central nervous system the extracellular matrix has important roles, e.g. supporting the extracellular space, controlling the tissue hydration, binding soluble factors and influencing their diffusion. The distribution of the extracellular matrix components in the brain has been mapped but data on the circumventricular organs (CVOs) is not available yet. The CVOs lack the blood-brain barrier and have relatively large perivascular spaces. The present study investigates tenascin-R and the lecticans: aggrecan, brevican, neurocan, and versican in the median eminence, the area postrema, the vascular organ of the lamina terminalis, the subfornical organ, the pineal body and the subcommissural organ of the rat applying immunohistochemical methods, and lectin histochemistry, using Wisteria floribunda agglutinin (WFA). The extracellular matrix components were found intensely expressed in the CVOs with two exceptions: aggrecan immunoreactivity visualized only neurons in the arcuate nucleus, and the subcommissural organ was not labeled with either WFA, or lecticans, or tenascin-R. The different labelings usually overlapped each other. The distribution of the extracellular matrix components marked the territories of the CVOs. Considering these we suppose that the extracellular matrix is essential in the maintenance of CVO functions providing the large extracellular space which is required for diffusion and other processes important in their chemosensitive and neurosecretory activities. The decrease of extracellular matrix beyond the border of the organs may contribute to the control of the diffusion of molecules from the CVOs into the surrounding brain substance.
Assuntos
Área Postrema/metabolismo , Proteínas da Matriz Extracelular/metabolismo , Hipotálamo/metabolismo , Eminência Mediana/metabolismo , Sistemas Neurossecretores/metabolismo , Agrecanas/metabolismo , Animais , Brevicam/metabolismo , Feminino , Masculino , Neurocam/metabolismo , Glândula Pineal/metabolismo , Ratos Wistar , Órgão Subcomissural/metabolismo , Órgão Subfornical/metabolismo , Tenascina/metabolismo , Versicanas/metabolismoRESUMO
The blood-brain barrier (BBB) is a barrier that prevents free access of blood-derived substances to the brain through the tight junctions and maintains a specialized brain environment. Circumventricular organs (CVOs) lack the typical BBB. The fenestrated vasculature of the sensory CVOs, including the organum vasculosum of the lamina terminalis (OVLT), subfornical organ (SFO) and area postrema (AP), allows parenchyma cells to sense a variety of blood-derived information, including osmotic ones. In the present study, we utilized immunohistochemistry to examine changes in the expression of NG2 and platelet-derived growth factor receptor beta (PDGFRB) in the OVLT, SFO and AP of adult mice during chronic osmotic stimulation. The expression of NG2 and PDGFRB was remarkably prominent in pericytes, although these angiogenesis-associated proteins are highly expressed at pericytes of developing immature vasculature. The chronic salt loading prominently increased the expression of NG2 in the OVLT and SFO and that of PDGFRB in the OVLT, SFO and AP. The vascular permeability of low-molecular-mass tracer fluorescein isothiocyanate was increased significantly by chronic salt loading in the OVLT and SFO but not AP. In conclusion, the present study demonstrates changes in pericyte expression of NG2 and PDGFRB and vascular permeability in the sensory CVOs by chronic osmotic stimulation, indicating active participation of the vascular system in osmotic homeostasis.
Assuntos
Antígenos/metabolismo , Área Postrema/metabolismo , Permeabilidade Capilar , Hipotálamo/metabolismo , Pericitos/metabolismo , Proteoglicanas/metabolismo , Receptor beta de Fator de Crescimento Derivado de Plaquetas/metabolismo , Órgão Subfornical/metabolismo , Animais , Antígenos/genética , Área Postrema/irrigação sanguínea , Área Postrema/citologia , Células Endoteliais/citologia , Células Endoteliais/metabolismo , Hipotálamo/irrigação sanguínea , Hipotálamo/citologia , Camundongos , Camundongos Endogâmicos C57BL , Osmorregulação , Pericitos/citologia , Proteoglicanas/genética , Receptor beta de Fator de Crescimento Derivado de Plaquetas/genética , Cloreto de Sódio/farmacologia , Órgão Subfornical/irrigação sanguínea , Órgão Subfornical/citologiaRESUMO
Calorie restriction (CR) has been shown to increase longevity and elicit many health promoting benefits including delaying immunosenescence and attenuating neurodegeneration in animal models of Alzheimer's disease and Parkinson's disease. CR also suppresses microglial activation following cortical injury and aging. We previously demonstrated that CR attenuates lipopolysaccharide (LPS)-induced fever and shifts hypothalamic signaling pathways to an anti-inflammatory bias; however, the effects of CR on LPS-induced microglial activation remain largely unexplored. The current study investigated regional changes in LPS-induced microglial activation in mice exposed to 50% CR for 28days. Immunohistochemistry was conducted to examine changes in ionized calcium-binding adapter molecule-1 (Iba1), a protein constitutively expressed by microglia, in a total of 27 brain regions involved in immunity, stress, and/or thermoregulation. Exposure to CR attenuated LPS-induced fever, and LPS-induced microglial activation in a subset of regions: the arcuate nucleus (ARC) and ventromedial nucleus of the hypothalamus (VMH) and the subfornical organ (SFO). Microglial activation in the ARC and VMH was positively correlated with body temperature. These data suggest that CR exerts effects on regionally specific populations of microglia; particularly, in appetite-sensing regions of the hypothalamus, and/or regions lacking a complete blood brain barrier, possibly through altered pro- and anti-inflammatory signaling in these regions.
Assuntos
Restrição Calórica , Hipotálamo/metabolismo , Microglia/metabolismo , Órgão Subfornical/metabolismo , Animais , Proteínas de Ligação ao Cálcio/metabolismo , Hipotálamo/efeitos dos fármacos , Lipopolissacarídeos/farmacologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Proteínas dos Microfilamentos/metabolismo , Microglia/efeitos dos fármacos , Órgão Subfornical/efeitos dos fármacosRESUMO
The circumventricular organs (CVOs), including the organum vasculosum of the lamina terminalis (OVLT), subfornical organ (SFO), and area postrema (AP) sense a variety of blood-borne molecules because they lack typical blood-brain barrier. Though a few signaling pathways are known, it is not known how endogenous ligands for transient receptor potential vanilloid receptor 1 ion channel (TRPV1) are sensed in the CVOs. In this study, we aimed to examine whether or not astrocytic TRPV1 senses directly blood-borne molecules in the OVLT, SFO, and AP of adult mice. The reverse transcription-polymerase chain reaction and Western analysis revealed the expression of TRPV1 in the CVOs. Confocal microscopic immunohistochemistry further showed that TRPV1 was localized prominently at thick cellular processes of astrocytes rather than fine cellular processes and cell bodies. TRPV1-expressing cellular processes of astrocytes surrounded the vasculature to constitute dense networks. The expression of TRPV1 was also found at neuronal dendrites but not somata in the CVOs. The intravenous administration of a TRPV1 agonist resiniferatoxin (RTX) prominently induced Fos expression at astrocytes in the OVLT, SFO, and AP and neurons in adjacent related nuclei of the median preoptic nuclei (MnPO) and nucleus of the solitary tract (Sol) of wild-type but not TRPV1-knockout mice. The intracerebroventricular infusion of RTX induced Fos expression at both astrocytes and neurons in the CVOs, MnPO, and Sol. Thus, this study demonstrates that blood-borne molecules are sensed directly by astrocytic TRPV1 of the CVOs in adult mammalians.
Assuntos
Astrócitos/metabolismo , Barreira Hematoencefálica/metabolismo , Encéfalo/metabolismo , Neurônios/metabolismo , Canais de Cátion TRPV/metabolismo , Animais , Encéfalo/irrigação sanguínea , Hipotálamo/irrigação sanguínea , Hipotálamo/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Órgão Subfornical/irrigação sanguínea , Órgão Subfornical/metabolismo , Canais de Cátion TRPV/genéticaRESUMO
Chronic intermittent hypoxia (CIH) models repetitive bouts of arterial hypoxemia that occur in humans suffering from obstructive sleep apnea. CIH has been linked to persistent activation of arterial chemoreceptors and the renin-angiotensin system, which have been linked to chronic elevations of sympathetic nerve activity (SNA) and mean arterial pressure (MAP). Because Fos and FosB are transcription factors involved in activator protein (AP)-1 driven central nervous system neuronal adaptations, this study determined if CIH causes increased Fos or FosB staining in brain regions that regulate SNA and autonomic function. Male Sprague Dawley rats were instrumented with telemetry transmitters for continuous recording of MAP and heart rate (HR). Rats were exposed to continuous normoxia (CON) or to CIH for 8 h/day for 7 days. CIH increased MAP by 7-10 mmHg without persistently affecting HR. A separate group of rats was killed 1 day after 7 days of CIH for immunohistochemistry. CIH did not increase Fos staining in any brain region examined. Staining for FosB/ΔFosB was increased in the organum vasculosum of the lamina terminalis (CON: 9 ± 1; CIH: 34 ± 3 cells/section), subfornical organ (CON: 7 ± 2; CIH: 31 ± 3), median preoptic nucleus (CON 15 ± 1; CIH: 38 ± 3), nucleus of the solitary tract (CON: 9 ± 2; CIH: 28 ± 4), A5 (CON: 3 ± 1; CIH: 10 ± 1), and rostral ventrolateral medulla (CON: 5 ± 1; CIH: 17 ± 2). In the paraventricular nucleus, FosB/ΔFosB staining was located mainly in the dorsal and medial parvocellular subnuclei. CIH did not increase FosB/ΔFosB staining in caudal ventrolateral medulla or supraoptic nucleus. These data indicate that CIH induces an increase in FosB/ΔFosB in autonomic nuclei and suggest that AP-1 transcriptional regulation may contribute to stable adaptive changes that support chronically elevated SNA.
Assuntos
Sistema Nervoso Autônomo/metabolismo , Pressão Sanguínea/fisiologia , Encéfalo/metabolismo , Hipertensão/fisiopatologia , Hipóxia/fisiopatologia , Proteínas Proto-Oncogênicas c-fos/metabolismo , Animais , Modelos Animais de Doenças , Frequência Cardíaca/fisiologia , Hipertensão/metabolismo , Hipotálamo/metabolismo , Masculino , Neurônios Aferentes/fisiologia , Núcleo Hipotalâmico Paraventricular/metabolismo , Área Pré-Óptica/metabolismo , Ratos , Ratos Sprague-Dawley , Núcleo Solitário/metabolismo , Órgão Subfornical/metabolismo , Sistema Nervoso Simpático/metabolismo , Sinapses/fisiologiaRESUMO
An increase in plasma ANG II causes neuronal activation in hypothalamic nuclei and a slow pressor response, presumably by increasing sympathetic drive. We evaluated whether the activation of a neuromodulatory pathway, involving aldosterone and "ouabain," is involved in these responses. In Wistar rats, the subcutaneous infusion of ANG II at 150 and 500 ng x kg(-1) x min(-1) gradually increased blood pressure up to 60 mmHg at the highest dose. ANG II at 500 ng x kg(-1) x min(-1) increased plasma ANG II by 4-fold, plasma aldosterone by 25-fold, and hypothalamic aldosterone by 3-fold. The intracerebroventricular infusion of an aldosterone synthase (AS) inhibitor prevented the ANG II-induced increase in hypothalamic aldosterone without affecting the increase in plasma aldosterone. Neuronal activity, as assessed by Fra-like immunoreactivity, increased transiently in the subfornical organ (SFO) but progressively in the paraventricular nucleus (PVN) and supraoptic nucleus (SON). The central infusion of the AS inhibitor or a mineralocorticoid receptor blocker markedly attenuated the ANG II-induced neuronal activation in the PVN but not in the SON. Pressor responses to ANG II at 150 ng x kg(-1) x min(-1) were abolished by an intracerebroventricular infusion of the AS inhibitor. Pressor responses to ANG II at 500 ng x kg(-1) x min(-1) were attenuated by the central infusion of the AS inhibitor or the mineralocorticoid receptor blocker by 70-80% and by Digibind (to bind "ouabain") by 50%. These results suggest a novel central nervous system mechanism for the ANG II-induced slow pressor response, i.e., circulating ANG II activates the SFO, leading to the direct activation of the PVN and SON, and, in addition, via aldosterone-dependent amplifying mechanisms, causes sustained activation of the PVN and thereby hypertension.
Assuntos
Aldosterona/metabolismo , Angiotensina II/sangue , Pressão Sanguínea , Encéfalo/metabolismo , Cardenolídeos/metabolismo , Neurônios/metabolismo , Sistema Renina-Angiotensina , Saponinas/metabolismo , Angiotensina II/administração & dosagem , Animais , Pressão Sanguínea/efeitos dos fármacos , Monitorização Ambulatorial da Pressão Arterial , Encéfalo/citologia , Encéfalo/efeitos dos fármacos , Citocromo P-450 CYP11B2/antagonistas & inibidores , Citocromo P-450 CYP11B2/metabolismo , Inibidores Enzimáticos/administração & dosagem , Fadrozol , Frequência Cardíaca , Hipotálamo/metabolismo , Imidazóis/administração & dosagem , Fragmentos Fab das Imunoglobulinas/administração & dosagem , Imuno-Histoquímica , Bombas de Infusão Implantáveis , Infusões Parenterais , Infusões Subcutâneas , Masculino , Antagonistas de Receptores de Mineralocorticoides/administração & dosagem , Vias Neurais/metabolismo , Neurônios/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-fos/metabolismo , Piridinas/administração & dosagem , Ratos , Ratos Wistar , Sistema Renina-Angiotensina/efeitos dos fármacos , Espironolactona/administração & dosagem , Órgão Subfornical/metabolismo , Telemetria , Fatores de TempoRESUMO
The spontaneously hypertensive rat (SHR) has an intense consumption of NaCl solution. Water deprivation (WD) followed by water intake to satiety induces partial rehydration (PR)-the WD-PR protocol-and sodium appetite. In the present work, WD produced similar water intake and no alterations in arterial pressure among spontaneously hypertensive rat (SHR), Wistar-Kyoto, and Holtzman strains. It also increased the number of cells with positive c-Fos immunoreactivity (Fos-IR) in the lamina terminalis and in the hypothalamic supraoptic (SON) and paraventricular (parvocellular, PVNp) nucleus in these strains. The WD and WD-PR produced similar alterations in all strains in serum osmolality and protein, plasma renin activity, and sodium balance. The SHR ingested about 10 times more 0.3 M NaCl than normotensives strains in the sodium appetite test that follows WD-PR. After WD-PR, the Fos-IR persisted, elevated in the lamina terminalis of all strains but notably in the subfornical organ of the SHR. The WD-PR reversed Fos-IR in the SON of all strains and in the PVNp of SHR. It induced Fos-IR in the area postrema and in the nucleus of the solitary tract (NTS), dorsal raphe, parabrachial (PBN), pre-locus coeruleus (pre-LC), suprachiasmatic, and central amygdalar nucleus of all strains. This effect was bigger in the caudal-NTS, pre-LC, and medial-PBN of SHRs. The results indicate that WD-PR increases cell activity in the forebrain and hindbrain areas that control sodium appetite in the rat. They also suggest that increased cell activity in facilitatory brain areas precedes the intense 0.3 M NaCl intake of the SHR in the sodium appetite test.
Assuntos
Apetite/fisiologia , Hipertensão/metabolismo , Hipotálamo/metabolismo , Proteínas Proto-Oncogênicas c-fos/metabolismo , Cloreto de Sódio na Dieta/farmacologia , Privação de Água/fisiologia , Tonsila do Cerebelo/metabolismo , Animais , Pressão Sanguínea/fisiologia , Ingestão de Líquidos/fisiologia , Ingestão de Alimentos/fisiologia , Eletrólitos/sangue , Eletrólitos/urina , Frequência Cardíaca/fisiologia , Hipertensão/fisiopatologia , Imuno-Histoquímica , Masculino , Área Pré-Óptica/metabolismo , Ratos , Ratos Endogâmicos SHR , Ratos Endogâmicos WKY , Ratos Sprague-Dawley , Órgão Subfornical/metabolismo , Equilíbrio Hidroeletrolítico/fisiologiaRESUMO
The up-regulation in the expression of mRNA or protein encoded by the c-fos gene is widely used as a marker of neuronal activation elicited by various stimuli. To facilitate the detection of activated neurons, we generated transgenic rats expressing a fusion gene consisting of c-fos coding sequences in frame with monomeric red fluorescent protein 1 (mRFP1) under the control of c-fos gene regulatory sequences (c-fos-mRFP1 rats). In c-fos-mRFP1 transgenic rats, 90 min after hypertonic saline ip administration, nuclear mRFP1 fluorescence was observed abundantly in brain regions known to be osmosensitive, namely the median preoptic nucleus, organum vasculosum lamina terminalis, supraoptic nucleus, paraventricular nucleus, and subfornical organ. Immunohistochemistry for Fos protein confirmed that the distribution of Fos-like immunoreactivity in nontransgenic rats was similar to those of mRFP1 fluorescence after ip administration of hypertonic saline in the transgenic rats. Several double-transgenic rats were obtained from matings between transgenic rats expressing an arginine vasopressin-enhanced green fluorescent protein fusion gene (AVP-eGFP rats) and c-fos-mRFP1 rats. In these double-transgenic rats, almost all eGFP neurons in the supraoptic nucleus and PVN expressed nuclear mRFP1 fluorescence 90 min after hypertonic saline administration. The c-fos-mRFP1 rats are a powerful tool that enables the facile identification of activated neurons in the nervous system. Furthermore, when combined with transgenes expressing another fluorophore under the control of cell-specific regulatory sequences, activation of specific neuronal cell types in response to physiological cues can be readily detected.
Assuntos
Arginina Vasopressina/metabolismo , Encéfalo/metabolismo , Neurônios/metabolismo , Proteínas Proto-Oncogênicas c-fos/metabolismo , Animais , Arginina Vasopressina/genética , Encéfalo/citologia , Feminino , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Hipotálamo/citologia , Hipotálamo/metabolismo , Imuno-Histoquímica , Proteínas Luminescentes/genética , Proteínas Luminescentes/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Microscopia Confocal , Microscopia de Fluorescência , Núcleo Hipotalâmico Paraventricular/citologia , Núcleo Hipotalâmico Paraventricular/metabolismo , Proteínas Proto-Oncogênicas c-fos/genética , Ratos , Ratos Transgênicos , Ratos Wistar , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Sequências Reguladoras de Ácido Nucleico/genética , Solução Salina Hipertônica/administração & dosagem , Solução Salina Hipertônica/farmacologia , Órgão Subfornical/citologia , Órgão Subfornical/metabolismo , Núcleo Supraóptico/citologia , Núcleo Supraóptico/metabolismo , Regulação para Cima/efeitos dos fármacos , Proteína Vermelha FluorescenteRESUMO
Previous studies have indicated that the renin-angiotensin-aldosterone system (RAAS) is implicated in the induction of sodium appetite in rats and that different dietary sodium intakes influence the mRNA expression of central and peripheral RAAS components. To determine whether dietary sodium deprivation activates regional brain neurons related to sodium appetite, and changes their gene expression of RAAS components of rats, the present study examined the c-Fos expression after chronic exposure to low sodium diet, and determined the relationship between plasma and brain angiotensin I (ANG I), angiotensin II (ANG II) and aldosterone (ALD) levels and the sodium ingestive behavior variations, as well as the effects of prolonged dietary sodium deprivation on ANG II type 1 (AT1) and ANG II type 2 (AT2) receptors and angiotensin-convertion enzyme (ACE) mRNA levels in the involved brain regions using the method of real-time polymerase chain reaction (PCR). Results showed that the Fos immunoreactivity (Fos-ir) expression in forebrain areas such as subfornical organ (SFO), paraventricular hypothalamic nuclei (PVN), supraoptic nucleus (SON) and organum vasculosum laminae terminalis (OVLT) all increased significantly and that the levels of ANG I, ANG II and ALD also increased in plasma and forebrain in rats fed with low sodium diet. In contrast, AT1, ACE mRNA in PVN, SON and OVLT decreased significantly in dietary sodium depleted rats, while AT2 mRNA expression did not change in the examined areas. These results suggest that many brain areas are activated by increased levels of plasma and/or brain ANG II and ALD, which underlies the elevated preference for hypertonic salt solution after prolonged exposure to low sodium diet, and that the regional AT1 and ACE mRNA are down-regulated after dietary sodium deprivation, which may be mediated by increased ANG II in plasma and/or brain tissue.
Assuntos
Encéfalo/metabolismo , Neurônios/metabolismo , Peptidil Dipeptidase A/genética , Receptor Tipo 1 de Angiotensina/genética , Sódio na Dieta/metabolismo , Sódio/deficiência , Aldosterona , Angiotensina I/metabolismo , Angiotensina II/metabolismo , Animais , Encéfalo/citologia , Regulação para Baixo/fisiologia , Regulação da Expressão Gênica/fisiologia , Hipotálamo/citologia , Hipotálamo/metabolismo , Hipotálamo Anterior/citologia , Hipotálamo Anterior/metabolismo , Masculino , Núcleo Hipotalâmico Paraventricular/citologia , Núcleo Hipotalâmico Paraventricular/metabolismo , Proteínas Proto-Oncogênicas c-fos/metabolismo , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Órgão Subfornical/citologia , Órgão Subfornical/metabolismoRESUMO
Circulating angiotensin II (ANGII) elicits water intake and activates the hypothalamic-pituitary-adrenal (HPA) axis by stimulating angiotensin type 1 receptors (AT1Rs) within circumventricular organs. The subfornical organ (SFO) and the organum vasculosum of the lamina terminalis (OVLT) are circumventricular organs that express AT1Rs that bind blood-borne ANGII and stimulate integrative and effector regions of the brain. The goal of these studies was to determine the contribution of AT1Rs within the SFO and OVLT to the water intake and HPA response to increased circulating ANGII. Antisense oligonucleotides directed against the AT1R [AT1R antisense (AT1R AS)] were administered into the OVLT or SFO. Quantitative receptor autoradiography confirmed that AT1R AS decreased ANGII binding in the SFO and OVLT compared with the scrambled sequence control but did not affect AT1R binding in other nuclei. Subsequently, water intake, ACTH, and corticosterone (CORT) were assessed after administration of isoproterenol, a beta-adrenergic agonist that decreases blood pressure and elevates circulating ANGII. Delivery of AT1R AS into the SFO attenuated water intake, ACTH, and CORT after isoproterenol, whereas similar treatment in the OVLT had no effect. To determine the specificity of this blunted drinking and HPA response, the same parameters were measured after treatment with hypertonic saline, a stimulus that induces drinking independently of ANGII. Delivery of AT1R AS into the SFO or OVLT had no effect on water intake, ACTH, or CORT after hypertonic saline. The results imply that AT1R within the SFO mediate drinking and HPA responses to stimuli that increase circulating ANGII.
Assuntos
Ingestão de Líquidos/efeitos dos fármacos , Sistema Hipotálamo-Hipofisário/efeitos dos fármacos , Isoproterenol/farmacologia , Sistema Hipófise-Suprarrenal/efeitos dos fármacos , Receptor Tipo 1 de Angiotensina/metabolismo , Órgão Subfornical/efeitos dos fármacos , Agonistas Adrenérgicos beta/farmacologia , Hormônio Adrenocorticotrópico/sangue , Angiotensina II/metabolismo , Bloqueadores do Receptor Tipo 1 de Angiotensina II/farmacologia , Animais , Corticosterona/sangue , Sistema Hipotálamo-Hipofisário/metabolismo , Sistema Hipotálamo-Hipofisário/fisiologia , Hipotálamo/efeitos dos fármacos , Hipotálamo/metabolismo , Hipotálamo/fisiologia , Masculino , Oligodesoxirribonucleotídeos Antissenso/farmacologia , Sistema Hipófise-Suprarrenal/metabolismo , Sistema Hipófise-Suprarrenal/fisiologia , Radioimunoensaio , Ratos , Ratos Sprague-Dawley , Receptor Tipo 1 de Angiotensina/fisiologia , Cloreto de Sódio/farmacologia , Órgão Subfornical/metabolismo , Órgão Subfornical/fisiologiaRESUMO
Stanniocalcin (STC-1), a 50 kDa glycoprotein hormone that regulates calcium/phosphate homeostasis in bony fish and mammals, has been shown to be expressed in central neurons and choroid plexus, and to exert a protective effect against hypercalcemic and hypoxic damage to neurons. Circumventricular organs are known to function in the regulation of ion and body fluid balance. Therefore, the possibility exists that STC-1 may be involved in the regulation of calcium/phosphate and fluid homeostasis through its actions on these central sites. In the present study, the distribution of STC-1 binding sites in forebrain circumventricular organs of the rat were investigated by in situ ligand binding using a stanniocalcin-alkaline phosphatase (STC-AP) fusion protein. Cells exhibiting STC-1 binding sites were found throughout the lamina terminalis. Dense cytoplasmic staining was observed predominantly within ependymal cells lining the anterior third ventricle region (AV3V), as well as cells of the choroid plexus. Additionally, neurons of the organum vasculosum of the lamina terminalis, the dorsal and ventral components of the median preoptic nucleus and the rostral aspects of the subfornical organ exhibited dense STC-1 cytoplasmic staining. STC-1 binding sites were also found in cells of the supraoptic nucleus, suprachiasmatic nucleus and anteroventral preoptic nucleus. These data suggest that STC-1 binding sites localized on the ependymal cells of the AV3V region and neurons of circumventricular organs may be associated with neuronal pathways involved in calcium/phosphate and fluid homeostasis.
Assuntos
Glicoproteínas/metabolismo , Hipotálamo/metabolismo , Animais , Autorradiografia , Sítios de Ligação/efeitos dos fármacos , Sítios de Ligação/fisiologia , Feminino , Masculino , Ratos , Ratos Wistar , Órgão Subfornical/metabolismo , Distribuição TecidualRESUMO
These studies examined the effects of increased dietary sodium on expression of Fos, the protein product of c-fos, in forebrain structures in the rat following intravenous infusion with angiotensin II (AngII). Animals were provided with either tap water (Tap) or isotonic saline solution (Iso) as their sole drinking fluid for 3-5 weeks prior to testing. Rats were then implanted with catheters in a femoral artery and vein. The following day, the conscious, unrestrained animals received iv infusion of either isotonic saline (Veh), AngII, or phenylephrine (Phen) for 2 h. Blood pressure and heart rate were monitored continuously throughout the procedure. Brains were subsequently processed for evaluation of Fos-like immunoreactivity (Fos-Li IR) in the organum vasculosum of the lamina terminalis (OVLT), the subfornical organ (SFO), and the median preoptic nucleus (MnPO). Fos-Li IR was significantly increased in the SFO and OVLT of animals consuming both Tap and Iso following AngII, but not Phen, compared to Veh infusions. Furthermore, Fos-Li IR in the MnPO was increased following AngII infusion in rats consuming a high sodium diet, but not in animals drinking Tap. These data suggest that increased dietary sodium sensitizes the MnPO neurons to excitatory input from brain areas responding to circulating AngII.
Assuntos
Angiotensina II/administração & dosagem , Hipotálamo/efeitos dos fármacos , Hipotálamo/metabolismo , Proteínas Proto-Oncogênicas c-fos/metabolismo , Sódio na Dieta/farmacologia , Angiotensina II/farmacologia , Animais , Infusões Intravenosas , Masculino , Fenilefrina/farmacologia , Área Pré-Óptica/efeitos dos fármacos , Área Pré-Óptica/metabolismo , Ratos , Ratos Sprague-Dawley , Cloreto de Sódio/farmacologia , Órgão Subfornical/efeitos dos fármacos , Órgão Subfornical/metabolismoRESUMO
The median preoptic nucleus (MnPO) in the lamina terminalis receives a prominent catecholaminergic innervation from the dorsomedial and ventrolateral medulla. The present investigation used whole cell patch-clamp recordings in rat brain slice preparations to evaluate the hypothesis that presynaptic adrenoceptors could modulate GABAergic inputs to MnPO neurons. Bath applications of norepinephrine (NE; 20-50 microM) induced a prolonged and reversible suppression of inhibitory postsynaptic currents (IPSCs) and reduced paired-pulse depression evoked by stimulation in the subfornical organ and organum vasculosum lamina terminalis. These events were not correlated with any observed changes in membrane conductance arising from NE activity at postsynaptic alpha(1)- or alpha(2)-adrenoceptors. Consistent with a role for presynaptic alpha(2)-adrenoceptors, responses were selectively mimicked by an alpha(2)-adrenoceptor agonist (UK-14304) and blockable with an alpha(2)-adrenoceptor antagonist (idazoxan). Although the alpha(1)-adrenoceptor agonist cirazoline and the alpha(1)-adrenoceptor antagonist prazosin were without effect on these evoked IPSCs, NE was noted to increase (via alpha(1)-adrenoceptors) or decrease (via alpha(2)-adrenoceptors) the frequency of spontaneous and tetrodotoxin-resistant miniature IPSCs. Collectively, these observations imply that both presynaptic and postsynaptic alpha(1)- and alpha(2)-adrenoceptors in MnPO are capable of selective modulation of rapid GABA(A) receptor-mediated inhibitory synaptic transmission along the lamina terminalis and therefore likely to exert a prominent influence in regulating cell excitability within the MnPO.
Assuntos
Hipotálamo/metabolismo , Área Pré-Óptica/metabolismo , Receptores Adrenérgicos alfa/metabolismo , Transdução de Sinais/fisiologia , Órgão Subfornical/metabolismo , Agonistas alfa-Adrenérgicos/farmacologia , Antagonistas Adrenérgicos alfa/farmacologia , Animais , Tartarato de Brimonidina , Células Cultivadas , Idazoxano/farmacologia , Imidazóis/farmacologia , Isoproterenol/farmacologia , Norepinefrina/metabolismo , Norepinefrina/farmacologia , Prazosina/farmacologia , Quinoxalinas/farmacologia , Ratos , Ratos WistarRESUMO
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.