RESUMO
Synaptic transmission is an essential process for neuron physiology. Such process is enabled in part due to modulation of neurotransmitter release. Adenosine is a synaptic modulator of neurotransmitter release in the Central Nervous System, including neurons of medulla oblongata, where several nuclei are involved with neurovegetative reflexes. Adenosine modulates different neurotransmitter systems in medulla oblongata, specially glutamate and noradrenaline in the nucleus tractussolitarii, which are involved in hypotensive responses. However, the intracellular mechanisms involved in this modulation remain unknown. The adenosine A2a receptor modulates neurotransmitter release by activating two cAMP protein effectors, the protein kinase A and the exchange protein activated by cAMP. Therefore, an in vitro approach (cultured cells) was carried out to evaluate modulation of neurotransmission by adenosine A2a receptor and the signaling intracellular pathway involved. Results show that the adenosine A2a receptor agonist, CGS 21680, increases neurotransmitter release, in particular, glutamate and noradrenaline and such response is mediated by protein kinase A activation, which in turn increased synapsin I phosphorylation. This suggests a mechanism of A2aR modulation of neurotransmitter release in cultured cells from medulla oblongata of Wistar rats and suggest that protein kinase A mediates this modulation of neurotransmitter release via synapsin I phosphorylation.
Assuntos
Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Neurônios/metabolismo , Neurotransmissores/metabolismo , Receptor A2A de Adenosina/metabolismo , Sinapsinas/metabolismo , Animais , Western Blotting , Células Cultivadas , Cromatografia Líquida de Alta Pressão , Humanos , Bulbo/metabolismo , Fosforilação , Ratos , Ratos Wistar , Transmissão Sináptica/fisiologiaRESUMO
Hypertension can result from neuronal network imbalance in areas of central nervous system that control blood pressure, such as the nucleus tractus solitarius (NTS). There are several neurotransmitters and neuromodulatory substances within the NTS, such as adenosine, which acts on purinoreceptors A(2a) (A(2a)R). The A(2a)R modulates neurotransmission in the NTS where its activation may induce decrease in blood pressure by different mechanisms. Nicotine is a molecule that crosses the hematoencephalic barrier and acts in several areas of central nervous system including the NTS, where it may interact with some neurotransmitter systems and contributes to the development of hypertension in subjects with genetic predisposition to this disease. In this study we first determined A(2a)R binding, protein, and mRNA expression in dorsomedial medulla oblongata of neonate normotensive (WKY) and spontaneously hypertensive rats (SHR). Subsequently, we analyzed the modulatory effects of nicotine on A(2a)R in cell culture in order to evaluate its possible involvement in the development of hypertension. Data showed a decreased A(2a)R binding and increased protein and mRNA expression in tissue sample and culture of dorsal brainstem from SHR compared with those from WKY rats at basal conditions. Moreover, nicotine modulated A(2a)R binding, protein, and mRNA expression in cells from both strains. Interestingly, nicotine decreased A(2a)R binding and increased protein levels, as well as, induced a differential modulation in A(2a)R mRNA expression. Results give us a clue about the mechanisms involved in the modulatory effects of nicotine on A(2a)R as well as hypothesize its possible contribution to the development of hypertension. In conclusion, we demonstrated that A(2a)R of SHR cells which differ from WKY and nicotine differentially modulates A(2a)R in dorsal brainstem cells of SHR and WKY.
Assuntos
Tronco Encefálico/efeitos dos fármacos , Tronco Encefálico/metabolismo , Nicotina/farmacologia , Agonistas Nicotínicos/farmacologia , Receptor A2A de Adenosina/metabolismo , Animais , Western Blotting , Células Cultivadas , Relação Dose-Resposta a Droga , Hipertensão/metabolismo , Bulbo/efeitos dos fármacos , Bulbo/metabolismo , Nicotina/administração & dosagem , Agonistas Nicotínicos/administração & dosagem , Reação em Cadeia da Polimerase , RNA Mensageiro/metabolismo , Ratos , Ratos Endogâmicos SHR , Ratos Endogâmicos WKY , Especificidade da Espécie , Fatores de TempoRESUMO
Adenosine acts in the nucleus tractus solitarii (NTS), one of the main brain sites related to cardiovascular control. In the present study we show that A(1) adenosine receptor (A(1R)) activation promotes an increase on alpha(2)-adrenoceptor (Alpha(2R)) binding in brainstem cell culture from newborn rats. We investigated the intracellular cascade involved in such modulatory process using different intracellular signaling molecule inhibitors as well as calcium chelators. Phospholipase C, protein kinase Ca(2+)-dependent, IP(3) receptor and intracellular calcium were shown to participate in A(1R)/Alpha(2R) interaction. In conclusion, this result might be important to understand the role of adenosine within the NTS regarding autonomic cardiovascular control.
Assuntos
Adenosina/metabolismo , Tronco Encefálico/metabolismo , Receptores Adrenérgicos alfa 2/metabolismo , Transdução de Sinais/fisiologia , Núcleo Solitário/metabolismo , Fosfolipases Tipo C/metabolismo , Adenosina/farmacologia , Animais , Vias Autônomas/efeitos dos fármacos , Vias Autônomas/metabolismo , Tronco Encefálico/efeitos dos fármacos , Cálcio/metabolismo , Sinalização do Cálcio/efeitos dos fármacos , Sinalização do Cálcio/fisiologia , Fenômenos Fisiológicos Cardiovasculares/efeitos dos fármacos , Células Cultivadas , Quelantes/farmacologia , Coração/inervação , Coração/fisiologia , Receptores de Inositol 1,4,5-Trifosfato/efeitos dos fármacos , Receptores de Inositol 1,4,5-Trifosfato/metabolismo , Ratos , Ratos Wistar , Receptores Adrenérgicos alfa 2/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Núcleo Solitário/efeitos dos fármacos , Fosfolipases Tipo C/efeitos dos fármacosRESUMO
The actions of neurotrophic factors i.e. basic fibroblast growth factor (bFGF, FGF-2) to neurons are related not only to neuronal development and maintenance but also to synaptic plasticity regarding neurotransmission. We analyzed here the levels of FGF-2 immunoreactivity in the nucleus tractus solitarii (NTS) of Wistar Kyoto rats in response to alterations of neuronal activity promoted by the stimulation of the baroreceptor reflex following an aortic coarctation-induced-hypertension. The FGF-2 immunoreactivity (IR) was found in the cytoplasm of the neurons and in the nuclei of the glial cells in the NTS. A large number of NTS neurons expressed FOS immunoreactivity 4 h after coarctation, as an indication of neuronal activity. Stereological methods showed an increased number of FGF-2 immunoreactive (ir) neuronal profiles (90%) and glial profiles (149%) in the NTS of the 72 h aortic coarctated rats. 1-week later, FGF-2 ir neurons were still increased (54%) but no change was found in the number of FGF-2 ir glial profiles. The double immunoperoxidase method revealed that the majority of the FGF-2 ir glial cells was glial fibrillary acidic protein (GFAP) positive astrocytes. GFAP immunohistochemistry showed an astroglial reaction at 72 h time-interval (55%) but not 1 week after stimulation. The number of the cresyl violet positive neurons and OX42 ir profiles (marker of activated microglia) in the NTS of coarctated rats were not different from control by 1 week and 1 month after the surgery, indicating a lack of NTS injury in this period following coarctation hypertension. FGF-2 may be an important neurotrophic factor in areas involved in the control of blood pressure. The increased FGF-2 IR in the NTS cells following neuronal stimulation may represent trophic and plastic adaptive responses in this nucleus in an autocrine/paracrine fashion.
Assuntos
Coartação Aórtica/patologia , Fator 2 de Crescimento de Fibroblastos/imunologia , Hipertensão/patologia , Núcleo Solitário/metabolismo , Animais , Pressão Sanguínea , Proteína Glial Fibrilar Ácida/metabolismo , Masculino , Peroxidase/metabolismo , Proteínas Proto-Oncogênicas c-fos/metabolismo , Ratos , Ratos WistarRESUMO
OBJECTIVE: Sexual dimorphism has been observed in arterial hypertension. Blood pressure levels are lower in female than in male spontaneously hypertensive rats (SHR). Angiotensin II (Ang II) plays a major role in the regulation of blood pressure. The aim of this study was to compare Ang II vascular reactivity and AT(1) and AT(2) receptor gene expression in female and male SHR. METHODS: SHR animals were divided into four groups: (I) male, (II) female in physiological estrus, (III) ovariectomized and (IV) ovariectomized treated with estrogen. Arterial blood pressure, AT(1) and AT(2) mRNA expression were determined. Ang II responses in aorta and mesenteric vessels were also evaluated. RESULTS: In female SHR, aorta and mesenteric microvessels were hyporeactive to Ang II in comparison to male SHR. In ovariectomized females, Ang II vasoconstriction was similar to that of males. Estrogen treatment abolished this difference. The mRNA expression for AT(1) was higher in aorta and mesenteric vessels from males than in females. In ovariectomized SHR, mRNA expression for AT(1) was comparable to that of males. Treatment with estrogen reversed the over expression observed. Whereas AT(2) gene expression did not differ, a lower ratio AT(1)/AT(2) was found in female than in male vessels. A higher mRNA expression for AT(1) was observed in kidney from male than in female. Ovariectomy resulted in up-regulation of this subtype receptor. Treatment with estrogen reversed the overexpression. AT(2) gene expression was higher in kidney from female than male SHR. Ovariectomy reduced AT(2) gene expression and estrogen treatment reversed the alteration observed in kidney. CONCLUSION: There is sexual dimorphism in vascular reactivity and in receptor gene expression to Ang II in SHR. We conclude that estrogen modulates AT(1) and AT(2) receptor gene expression and that this might explain at least partially the lower blood pressure observed in female SHR.