RESUMO
Pineal gland secretes the hormone melatonin at night with a circadian rhythm. The synthesis and secretion of melatonin are stimulated at night by norepinephrine released by sympathetic postganglionic neurons projecting from the superior cervical ganglia. Norepinephrine simultaneously activates α- and ß-adrenoceptors, triggering melatonin synthesis.To study the regulation of melatonin production and secretion, it is very convenient to use an ex vivo preparation. Thus, it is possible to keep intact pineal glands in culture and to study the actions of agonists, antagonists, modulators, toxic agents, etc., in melatonin synthesis. Artificial melatonin synthesis stimulation in vitro is usually achieved by using a ß-adrenergic agonist alone or in association with an α-adrenergic agonist. In this chapter, the methodology of cultured pineal glands will be described. Several papers were published by our group using this methodology, approaching the role played in melatonin synthesis control by angiotensin II and IV, insulin, glutamate, voltage-gated calcium channels, anhydroecgonine methyl ester (AEME, crack-cocaine product), monosodium glutamate (MSG), signaling pathways like NFkB, pathophysiological conditions like diabetes, etc.
Assuntos
Cocaína , Insulinas , Melatonina , Glândula Pineal , Agonistas alfa-Adrenérgicos/metabolismo , Agonistas alfa-Adrenérgicos/farmacologia , Agonistas Adrenérgicos beta/farmacologia , Angiotensina II/metabolismo , Canais de Cálcio/metabolismo , Ritmo Circadiano/fisiologia , Melatonina/metabolismo , Norepinefrina , Glândula Pineal/metabolismo , Receptores Adrenérgicos beta/metabolismo , Glutamato de SódioRESUMO
Classical actions of the neurotrophin family are related to cellular survival and differentiation. Moreover, acute effects of neurotrophins have been reported. Although neurotrophins effects on synaptic transmission at central nervous system level have been largely studied, acute effects of neurotrophins on hypothalamic noradrenergic transmission are still poorly understood. Thus, we have studied the effects of the neurotrophin family members nerve growth factor (NGF), brain derived neurotrophic factor (BDNF) and neurotrophin-4 (NT-4) on norepinephrine (NE) neuronal uptake and its evoked release, as well as the receptor and the intracellular pathways involved in these processes in rat hypothalamus. Present results indicate that BDNF increased NE uptake and decreased its evoked release through a mechanism that involve Trk B receptor and phospholipase C. Moreover, NT-4, also through the Trk B receptor, decreased NE uptake and its evoked release by activating phosphatidylinositol 3-OH-kinase. These effects were observed in whole hypothalamus as well as in the anterior hypothalamic zone. On the other hand, NGF did not modify noradrenergic transmission. In conclusion, we showed for the first time that BDNF and NT-4 activate two different intracellular signalling pathways through a Trk B receptor dependent mechanism. Furthermore, present findings support the hypothesis that BDNF and NT-4 acutely applied, could be considered as modulators of noradrenergic transmission and thus may regulate hypothalamic physiological as well as pathophysiological responses.
Assuntos
Agonistas alfa-Adrenérgicos/metabolismo , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Hipotálamo/metabolismo , Fatores de Crescimento Neural/metabolismo , Norepinefrina/metabolismo , Transdução de Sinais/fisiologia , Animais , Inibidores Enzimáticos/metabolismo , Feminino , Hipotálamo/anatomia & histologia , Masculino , Fator de Crescimento Neural/metabolismo , Neurônios/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Inibidores de Fosfoinositídeo-3 Quinase , Ratos , Ratos Sprague-Dawley , Receptor trkB/metabolismo , Transmissão Sináptica/fisiologia , Fosfolipases Tipo C/antagonistas & inibidores , Fosfolipases Tipo C/metabolismoRESUMO
The biological effects of catecholamines in mammalian pigment cells are poorly understood. Our previous results showed the presence of alpha(1)-adrenoceptors in SK-Mel 23 human melanoma cells. The aims of this work were to (1) characterize catecholamine effects on proliferation, tyrosinase activity and expression, (2) identify the alpha(1)-adrenoceptor subtypes, and (3) verify whether chronic norepinephrine (NE) treatment modified the types and/or pharmacological characteristics of adrenoceptors present in SK-Mel 23 human melanoma cells. Cells treated with the alpha(1)-adrenergic agonist, phenylephrine (PHE, 10(-5) or 10(-4) M), for 24-72 h, exhibited decreased cell proliferation and enhanced tyrosinase activity, but unaltered tyrosinase expression as compared with the control. The proliferation and tyrosinase activity responses were inhibited by the alpha(1)-adrenergic antagonist prazosin, suggesting they were evoked by alpha(1)-adrenoceptors. The presence of actinomycin D, a transcription inhibitor, did not diminish PHE-induced effects. RT-PCR assays, followed by cloning and sequencing, demonstrated the presence of alpha(1A)- and alpha(1B)-adrenoceptor subtypes. NE-treated cells (24 or 72 h) were used in competition assays, and showed no significant change in the competition curves of alpha(1)-adrenoceptors as compared with control curves. Other adrenoceptor subtypes were not identified in these cells, and NE pretreatment did not induce their expression. In conclusion, the activation of SK-Mel 23 human melanoma alpha(1)-adrenoceptors elicit biological effects, such as proliferation decrease and tyrosinase activity increase. Desensitization or expression of other adrenoceptor subtypes after chronic NE treatment were not observed.
Assuntos
Agonistas alfa-Adrenérgicos/farmacologia , Melanoma , Norepinefrina/farmacologia , Receptores Adrenérgicos alfa 1/metabolismo , Neoplasias Cutâneas , Agonistas de Receptores Adrenérgicos alfa 1 , Agonistas alfa-Adrenérgicos/metabolismo , Ligação Competitiva , Divisão Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Expressão Gênica , Humanos , Monofenol Mono-Oxigenase/metabolismo , Norepinefrina/metabolismo , Fenilefrina/metabolismo , Fenilefrina/farmacologia , Receptores Adrenérgicos alfa 1/genéticaRESUMO
1. Age-related changes in the reactivity of postsynaptic alpha-adrenoceptors of isolated portions (epididymal and prostatic) and in whole vas deferens have been studied using 4, 12 and 20 month-old rats. 2. The pD2 values for adrenaline-induced contractions were reduced in the epididymal portion and whole vas deferens of middle-aged and old animals, but not in the prostatic portion. No age related change to phenylephrine or clonidine sensitivity was observed. 3. pA2 values of prazosin and yohimbine were not changed by aging in any preparation. Phentolamine pA2 values were reduced in the epididymal portion and in the whole vas deferens when adrenaline, but not when phenylephrine concentration-response curves were displaced by the antagonist. The mean pA2 value of yohimbine (6.78) indicates that this antagonist blocks alpha(1)-adrenoceptors in the rat vas deferens. 4. The data presented here suggest that age-related decreases in the sensitivity to adrenaline and phentolamine (when measured by displacing adrenaline concentration-effect curves) in the whole vas deferens are probably due to a variation in the proportion of alpha(1)-adrenoceptor subtypes in the epididymal portion of the rat vas deferens.
Assuntos
Envelhecimento/fisiologia , Epididimo/ultraestrutura , Próstata/ultraestrutura , Receptores Adrenérgicos alfa/fisiologia , Ducto Deferente/ultraestrutura , Agonistas alfa-Adrenérgicos/metabolismo , Agonistas alfa-Adrenérgicos/farmacologia , Antagonistas Adrenérgicos alfa/metabolismo , Antagonistas Adrenérgicos alfa/farmacologia , Envelhecimento/metabolismo , Animais , Epididimo/efeitos dos fármacos , Epinefrina/farmacologia , Cinética , Masculino , Contração Muscular/efeitos dos fármacos , Contração Muscular/fisiologia , Músculo Liso/efeitos dos fármacos , Músculo Liso/ultraestrutura , Fenilefrina/farmacologia , Próstata/efeitos dos fármacos , Ratos , Ratos Wistar , Receptores Adrenérgicos alfa/metabolismo , Ducto Deferente/efeitos dos fármacosAssuntos
Sacarose Alimentar/farmacologia , Hipotálamo/química , Receptores Adrenérgicos alfa 2/metabolismo , Agonistas de Receptores Adrenérgicos alfa 2 , Agonistas alfa-Adrenérgicos/metabolismo , Agonistas alfa-Adrenérgicos/farmacologia , Animais , Ligação Competitiva/fisiologia , Peso Corporal , Clonidina/metabolismo , Clonidina/farmacologia , Hipotálamo/efeitos dos fármacos , Masculino , Ratos , Ratos Wistar , TrítioRESUMO
Se estudió el efecto de agonistas alfa y beta sobre los mecanismos de transporte de electrolitos a nivel de las células de túbulo proximal en rebanadas de corteza renal de cobayo, a saber: el mecanismo que intercambia Na+ por K+ y el mecanismo que expulsa Na+ acompañado de Cl- y agua. Los resultados fueron los siguientes: 1) Todas las drogas simpaticomiméticas, indistintamente de su potencia relativa para estimular un tipo de receptor, estimularon la expulsión de Cl- y agua, mientras que inhibieron parcialmente la ganancia de K+. 2) Los effectos anteriormente descritos, fueron mimetizados por un aumento en el gradiente de Ca2+ externo y por la teofilina, la cual actuó en ausencia de Ca2+ externo. 3) En ausencia de Ca2+ externo, la norepinefrina (agonista alfa) fue incapaz de producir efecto alguno; por el contrario, el isoproterenol (agonista beta) no necesitó Ca2+ externo para producir su efecto. 4) Con los resultados presentados, se puede sugerir que los agonistas alfa pudieran afectar los movimientos activos de iones en este tejido a través de Ca2+ extracelular mientras que los agonistas beta lo harían mediante la liberación de Ca2+ endógeno