RESUMO
Although the existence of central responses to inflammatory injuries was already reported, the existence of hypothalamic amino acid responses has been less explored. The present study was designed to characterize the 24-h changes in mediobasal hypothalamic excitatory and inhibitory amino acid neurotransmitter contents and to analyze the effect of Freund's complete adjuvant administration on these patterns. Also the effects of the immunosuppressant drug Cyclosporine was studied. The content of aspartate, glutamate, glutamine, GABA and taurine was measured by HPLC with fluorimetric detection. The results show the existence of specific daily rhythms of aspartate, glutamate, glutamine, GABA and taurine contents in the mediobasal hypothalamus of control rats. Maxima for these amino acids was found at midnight, although another peak of lesser magnitude, occurred during the light phase of the photoperiod, except for TAU in which both peaks were of similar magnitude. Freund's complete adjuvant administration did not modify the 24-h pattern of any amino acid studied. It reduced the midnight peak of glutamate, glutamine and GABA and increased that of taurine. Moreover, it increased and extended the midday peak of glutamate. Besides, Freund's adjuvant did not modify aspartate content at any time point studied. Cyclosporine pretreatment did not prevent the inhibitory effects of Freund's complete adjuvant on glutamate, glutamine and GABA midnight peaks. However, the drug blocked the increase in the content of taurine at midnight and increased its midday peak. Moreover, cyclosporine administration abolished the variations of ASP during the scotophase, as compared to control animals and shift delayed both peaks of glutamate. The results indicate the existence of a significant effect of immune-mediated inflammatory response of the mediobasal hypothalamic amino acids studied, at an early phase after Freund's adjuvant administration, and that these changes were partially sensitive to the immunosuppression induced by cyclosporine.
Assuntos
Aminoácidos/metabolismo , Ritmo Circadiano/fisiologia , Ciclosporina/farmacologia , Adjuvante de Freund/farmacologia , Hipotálamo Médio/metabolismo , Animais , Injeções , Masculino , Concentração Osmolar , Ratos , Ratos WistarRESUMO
Substance P (SP) may participate as a paracrine and/or autocrine factor in the regulation of anterior pituitary function. This project studied the effect of TRH on SP content and release from anterior pituitary and the role of SP in TRH-induced prolactin release. TRH (10(-7) M), but not vasoactive intestinal polypeptide (VIP), increased immunoreactive-SP (ir-SP) content and release from male rat anterior pituitary in vitro. An anti-prolactin serum also increased ir-SP release and content. In order to determine whether intrapituitary SP participates in TRH-induced prolactin release, anterior pituitaries were incubated with TRH (10(-7) M) and either WIN 62,577, a specific antagonist of the NK1 receptor, or a specific anti-SP serum. Both WIN 62,577 (10(-8) and 10(-7) M) and the anti-SP serum (1:250) blocked TRH-induced prolactin release. In order to study the interaction between TRH and SP on prolactin release, anterior pituitaries were incubated with either TRH (10(-7) M) or SP, or with both peptides. SP (10(-7) and 10(-6) M) by itself stimulated prolactin release. While 10(-7) M SP did not modify the TRH effect, 10(-6) M SP reduced TRH-stimulated prolactin release. SP (10(-5) M) alone failed to stimulate prolactin release and markedly decreased TRH-induced prolactin release. The present study shows that TRH stimulates ir-SP release and increases ir-SP content in the anterior pituitary. Our data also suggest that SP may act as a modulator of TRH effect on prolactin secretion by a paracrine mechanism.
Assuntos
Comunicação Parácrina , Adeno-Hipófise/metabolismo , Prolactina/metabolismo , Substância P/farmacologia , Hormônio Liberador de Tireotropina/farmacologia , Análise de Variância , Androstenos/farmacologia , Animais , Benzimidazóis/farmacologia , Relação Dose-Resposta a Droga , Sinergismo Farmacológico , Soros Imunes/farmacologia , Antagonistas dos Receptores de Neurocinina-1 , Técnicas de Cultura de Órgãos , Adeno-Hipófise/química , Adeno-Hipófise/efeitos dos fármacos , Prolactina/análise , Ratos , Ratos Wistar , Estimulação Química , Substância P/análise , Substância P/metabolismo , Hormônio Liberador de Tireotropina/metabolismo , Peptídeo Intestinal Vasoativo/farmacologiaRESUMO
Nitric oxide (NO) affects cyclooxygenase (COX) and lipooxygenase (LOX) activities in several tissues. The aim of this study was to investigate the effect of NO on the AA metabolism in the anterior pituitary. LOX and COX products from anterior pituitaries of Wistar male rats were determined by [14C]-AA radioconversion method. Sodium nitroprusside (NP, 0.5 mM) and DETA NONOate (1 mM), NO donors, decreased 5-hydroxy-5,8,11,14-eicosatetraenoic acid (5-HETE) synthesis (P<0.05), effects that were reversed by hemoglobin. L-arginine also inhibited LOX activity. To the contrary, the inhibition of NO synthase by L-NAME (0.5 mM) or aminoguanidine (0.5 mM) increased 5-HETE production (P<0.05). COX activity was slightly stimulated by NP and L-arginine. However, DETA NONOate induced a stimulation of the synthesis of all prostanoids (P<0.05), this effect being reversed by hemoglobin. Neither NOS inhibitors nor hemoglobin modified basal prostanoids synthesis. These results indicate that NO inhibits LOX activity and stimulates COX activity in the anterior pituitary gland. The inhibition of LOX by NO may be another mechanism involved in the effects of NO on hormone release in the anterior pituitary.
Assuntos
Ácido Araquidônico/metabolismo , Óxido Nítrico/fisiologia , Adeno-Hipófise/metabolismo , Animais , Relação Dose-Resposta a Droga , Sequestradores de Radicais Livres/farmacologia , Ácidos Hidroxieicosatetraenoicos/biossíntese , Lipoxigenase/efeitos dos fármacos , Lipoxigenase/metabolismo , Masculino , Óxido Nítrico/farmacologia , Prostaglandina-Endoperóxido Sintases/efeitos dos fármacos , Prostaglandina-Endoperóxido Sintases/metabolismo , Ratos , Ratos WistarRESUMO
The effect of nitric oxide donors on intracellular calcium concentration [Ca2+]i was studied in anterior pituitary cells using ratiometric FURA 2 fluorescence measurements. Sodium nitroprusside (NP) induced a transient decrease in [Ca2+]i, after which [Ca2+]i returned to, or even increased over basal values. S-Nitroso glutathione (GSNO) induced a similar decrease. NP also inhibited high [Ca2+]i achieved by depolarization with 25 mM K+. The inhibitory effect of NP was partially blunted by pretreatment with methoxy-verapamil, and in calcium free buffer, and was not altered by thapsigargin. Interestingly, in calcium free buffer there was a significant stimulatory effect of NP, which was partially blunted by thapsigargin. We conclude that NO donors modify [Ca2+]i in anterior pituitary cells. The action is biphasic, with an initial decrease in [Ca2+]i probably related to a decrease of Ca2+ influx through VDCC, and an increase evidenced in calcium free buffer in which the inhibitory component is absent, and partially depends on thapsigargin sensitive calcium stores.
Assuntos
Cálcio/metabolismo , Doadores de Óxido Nítrico/farmacologia , Adeno-Hipófise/metabolismo , Animais , Soluções Tampão , Cálcio/administração & dosagem , Bloqueadores dos Canais de Cálcio/farmacologia , Galopamil/farmacologia , Glutationa/análogos & derivados , Glutationa/farmacologia , Líquido Intracelular/metabolismo , Cinética , Masculino , Nitroprussiato/farmacologia , Compostos Nitrosos/farmacologia , Potássio/farmacologia , Ratos , Ratos Wistar , S-Nitrosoglutationa , Tapsigargina/farmacologiaRESUMO
OBJECTIVE: In order to determine the mechanism by which nitric oxide (NO) inhibits prolactin release, we investigated the participation of cGMP-dependent cAMP-phosphodiesterases (PDEs) and protein kinase G (PKG) in this effect of NO. METHODS: Anterior pituitary glands of male rats were incubated with inhibitors of PDE and PKG with or without sodium nitroprusside (NP). Prolactin release, and cAMP and cGMP concentrations were determined by RIA. RESULTS AND CONCLUSIONS: The inhibitory effect of NP (0.5 mmol/l) on prolactin release and cAMP concentration was blocked by EHNA (10(-4)mol/l) and HL-725 (10(-4)mol/l), inhibitors of cGMP-stimulated cAMP-PDE (PDE2). 8-Br-cGMP (10(-4) and 10(-3)mol/l), which mimics cGMP as a mediator of NP effects on prolactin release, also decreased cAMP concentration. Zaprinast (10(-4)mol/l), a selective inhibitor of specific cGMP-PDE (PDE5), potentiated the NP effect on cAMP concentration. Rp-8-[(4-chlorophenyl)thio]-cGMP triethylamine (Rp-8-cGMP, 10(-7)-10(-6)mol/l), an inhibitor of PKG, reversed the effect of NP on prolactin release. The present study suggests that several mechanisms are involved in the inhibitory effect of NO on prolactin release. The activation of PDE2 by cGMP may mediate the inhibitory effect of NO on cAMP concentration and therefore on prolactin release. NO-activated PKG may also be participating in the inhibitory effect of NO on prolactin release.
Assuntos
Óxido Nítrico/farmacologia , Diester Fosfórico Hidrolases/metabolismo , Adeno-Hipófise/efeitos dos fármacos , Adeno-Hipófise/metabolismo , Prolactina/metabolismo , Proteínas Quinases/metabolismo , 3',5'-AMP Cíclico Fosfodiesterases/metabolismo , 3',5'-GMP Cíclico Fosfodiesterases/antagonistas & inibidores , 3',5'-GMP Cíclico Fosfodiesterases/metabolismo , Animais , AMP Cíclico/metabolismo , GMP Cíclico/análogos & derivados , GMP Cíclico/metabolismo , GMP Cíclico/farmacologia , Proteínas Quinases Dependentes de GMP Cíclico , Sinergismo Farmacológico , Ativação Enzimática/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Masculino , Nitroprussiato/farmacologia , Inibidores de Fosfodiesterase/farmacologia , Purinonas/farmacologia , Ratos , Ratos WistarRESUMO
The effect of prolactin on the activity of GABA-related enzymes and GABA concentrations were studied in hypothalamic slices incubated in vitro. After short periods of incubation (up to 40 min), prolactin (0.25 micrograms/ml) added to the incubation medium produced a significant increase (21% at 20 min of incubation) in glutamic acid decarboxylase (GAD) activity in the hypothalamic slices. A higher concentration of prolactin (1.0 micrograms/ml) produced a slight but significant decrease (8% at 20 min of incubation) in hypothalamic GAD activity. However, after longer periods of incubation (over 8 hr), both doses of prolactin induced a sustained increase in hypothalmic GAD activity, a response which depends upon protein synthesis. No changes were observed in GABA-transaminase (GABA-T) activity of hypothalamic slices incubated in the presence of prolactin. Prolactin decreased GABA concentration in the hypothalami incubated for 10 hr and, at the same time, increased GABA release into the medium. These results indicate that prolactin modifies the synthesis and release of hypothalmic GABA and suggest the existence of a feedback mechanism that prolactin may exert directly at the hypothalamic level.
Assuntos
Glutamato Descarboxilase/metabolismo , Hipotálamo/efeitos dos fármacos , Prolactina/farmacologia , Ácido gama-Aminobutírico/metabolismo , Animais , Técnicas de Cultura , Relação Dose-Resposta a Droga , Masculino , Ratos , Ratos EndogâmicosRESUMO
The anterior pituitary gland is a site of nitric oxide (NO) production and action, suggesting a local regulatory function. We recently reported that NO inhibits in vitro prolactin release. The aim of the present study was to establish the mechanism of action of NO on prolactin release and to determine whether NO is involved in the inhibitory effect of GABA on prolactin release. Since NO exerts its action through cGMP by activating guanylate cyclase in different tissues, we examined the effect of sodium nitroprusside (NP), a NO releaser, on intrapituitary cGMP levels. Incubation of anterior pituitary glands with 0.5 mM NP 4-fold increased intrapituitary cGMP content, but decreased intrapituitary cAMP levels. In addition, we studied the effect of NP on prolactin release in the presence of LY 83583, an inhibitor of guanylate cyclase activity and 3-isobutyl-1-methylxanthine (IBMX), an inhibitor of phosphodiesterase activity. 10 microM LY 83583 and 0.5 mM IBMX blocked the inhibitory effect of NP on prolactin release. (10(-3) M) 8Br-cGMP, an analogue of cGMP, mimicked the effect of NP on prolactin release. On the other hand, NO seems to be involved in the inhibitory effect of GABA on prolactin release since hemoglobin, a scavenger of NO, and Nw-nitro-L-arginine methyl ester, an inhibitor of NO synthase (NOS), blocked the pituitary response to GABA. Moreover, GABA (10(-6) M) stimulated NOS activity by almost 50%. GABA increased intrapituitary cGMP levels and decreased cAMP. Dopamine stimulated NOS activity weakly. These observations suggest that NO, acting through the guanylate cyclase-cGMP pathway, inhibits prolactin secretion. In addition, NO may be involved in the inhibitory effect of GABA and dopamine on prolactin release.
Assuntos
GMP Cíclico/metabolismo , Dopamina/farmacologia , Óxido Nítrico/metabolismo , Prolactina/metabolismo , Ácido gama-Aminobutírico/farmacologia , 1-Metil-3-Isobutilxantina/farmacologia , Aminoquinolinas/farmacologia , Animais , AMP Cíclico/metabolismo , GMP Cíclico/análogos & derivados , GMP Cíclico/farmacologia , Inibidores Enzimáticos/farmacologia , Hemoglobinas/farmacologia , Masculino , NG-Nitroarginina Metil Éster/farmacologia , Neurotransmissores/metabolismo , Inibidores de Fosfodiesterase/farmacologia , Hipófise/efeitos dos fármacos , Hipófise/metabolismo , Prolactina/efeitos dos fármacos , Ratos , Ratos WistarRESUMO
Abstract The effects of chronic administration of sex steroids on the content of vasoactive intestinal peptide (VIP) in the mediobasal hypothalamus and anterior pituitary were studied in adult rats. Gonadectomy had no effect on VIP concentration in the mediobasal hypothalamus or anterior pituitary gland. Estradiol benzoate (1 mug/100 g body wt/day) administered for 10 days decreased mediobasal hypothalamus VIP concentration of ovariectomized rats whereas it produced no change in mediobasal hypothalamus VIP content of orchidectomized rats. Testosterone propionate (100 mug/100 g body wt/day) administration decreased mediobasal hypothalamus VIP content in both sexes. Estradiol administration caused an increase whereas testosterone treatment resulted in a decrease in anterior pituitary VIP levels in both sexes. The effect of chronic administration of the sex steroids on VIP release from the mediobasal hypothalamus and anterior pituitary was also investigated. Estradiol increased evoked VIP release from the mediobasal hypothalamus and decreased mediobasal hypothalamus VIP content whereas testosterone decreased both mediobasal hypothalamus release and content. Chronic treatment with estradiol enhanced anterior pituitary VIP release and content while testosterone decreased both parameters studied. The data indicate that anterior pituitary VIP content is under the control of gonadal hormones and that the increased anterior pituitary VIP found after estradiol administration may be due to an augmented release from the mediobasal hypothalamus and probably an increase in anterior pituitary VIP synthesis.
RESUMO
A study was performed on the effect of ethanol on the basal and K+-evoked efflux of endogenous GABA from rat hypothalamic fragments. The amount of GABA present in the medium and in the tissue was measured by radioreceptor assay. In vitro addition of ethanol (50 and 100 mM) enhanced the K+-evoked efflux of GABA in a Ca++-dependent manner, and increased tissue GABA content. Since K+-evoked outflow induced by ethanol was not affected by the presence of nipecotic acid, ethanol appears to alter the uptake of endogenous GABA. An inhibitory effect of ethanol on 3H-GABA uptake was observed under K+ depolarization. On the other hand, acute ethanol administration produced a decrease in basal and K+-evoked efflux from hypothalamic fragments and in tissue GABA concentration. Changes in GABA efflux may lie behind some of the neuropharmacological effects of ethanol.
Assuntos
Etanol/farmacologia , Hipotálamo/metabolismo , Prolina/análogos & derivados , Ácido gama-Aminobutírico/metabolismo , Animais , Cálcio/fisiologia , Glutamato Descarboxilase/metabolismo , Hipotálamo/efeitos dos fármacos , Masculino , Ácidos Nipecóticos/farmacologia , Potássio/farmacologia , Ensaio Radioligante , Ratos , Ratos EndogâmicosRESUMO
In order to examine the role of hypothalamic SP in the feedback regulation of prolactin, we studied the effect of prolactin and dopamine on SP concentration and release, and the effect of SP on dopamine release. Hypothalamic fragments from male Wistar rats were incubated in the presence of prolactin, dopamine or SP under basal and K(+)-stimulated conditions. SP (10(-7) M) stimulated dopamine release, while dopamine (10(-7) M) decreased SP content and release. Prolactin (100 ng ml-1) increased SP content and release. An increase in hypothalamic SP content was also found during suckling. In addition, a specific antagonist for SP, Win 62,577, blocked the effect of prolactin and dopamine release. These results show an interaction between SP and dopamine at the hypothalamic level and suggest that SP could mediate the feedback action of prolactin on dopamine release.
Assuntos
Dopamina/metabolismo , Hipotálamo/fisiologia , Prolactina/farmacologia , Substância P/fisiologia , Androstenos/farmacologia , Animais , Benzimidazóis/farmacologia , Hipotálamo/efeitos dos fármacos , Hipotálamo/metabolismo , Masculino , Antagonistas dos Receptores de Neurocinina-1 , Ratos , Ratos WistarRESUMO
The effects of the acute injection of a rabbit anti-substance P serum (ASPS) were studied in normal rats and rats with hyperprolactinemia induced by 5-hydroxytryptophan and estradiol given as a short or chronic treatment. The anti-substance P serum decreased the release of prolactin induced by 5-hydroxytryptophan when this serotonin precursor was injected 24 h, but not 1 h, after the administration of the antiserum. ASPS reduced the hyperprolactinemia induced by short and chronic treatment with estradiol in castrated rats. This effect was observed 24 h after the injection of the antiserum. On the other hand, the injection of ASPS induced a significant decrease in LH levels in serum of intact male rats injected with 5-hydroxytryptophan 24 h after ASPS, and in castrated rats treated with short-term and chronic administration of estradiol, 24 h after the injection of the antiserum. These results suggest that substance P may have a role in the control of prolactin secretion and could play a part in the hyperprolactinemic effects of estradiol. On the other hand, substance P, under certain circumstances, may stimulate LH release.
Assuntos
Hormônio Foliculoestimulante/metabolismo , Hiperprolactinemia/fisiopatologia , Soros Imunes , Hormônio Luteinizante/metabolismo , Prolactina/metabolismo , Substância P/fisiologia , 5-Hidroxitriptofano/farmacologia , Animais , Estradiol , Feminino , Hormônio Foliculoestimulante/sangue , Hiperprolactinemia/induzido quimicamente , Hormônio Luteinizante/sangue , Masculino , Ovariectomia , Adeno-Hipófise/efeitos dos fármacos , Adeno-Hipófise/metabolismo , Prolactina/sangue , Ratos , Ratos Endogâmicos , Serotonina , Substância P/imunologiaRESUMO
The effect of the administration of a rabbit anti-substance P serum (ASPS) was studied in rats receiving an acute injection of ethanol. ASPS lowered serum prolactin levels and reduced the hyperprolactinemia induced by ethanol. ASPS also decreased LH serum levels in both saline- and ethanol-treated rats. The effect of ethanol on the concentration of substance P-like immunoreactivity (SP-LI) in the mediobasal hypothalamus and the anterior pituitary gland was also investigated. Ethanol reduced SP-LI in the mediobasal hypothalamus but increased it in the anterior pituitary gland. The presence of ethanol (50 mM) did not affect the K(+)-evoked release of SP-LI from either mediobasal hypothalamus or anterior pituitary gland, though it increased the SP-LI concentration remaining in this gland. These results indicate that ethanol increases the content of SP-LI in the anterior pituitary gland and suggest that substance P may be involved in the prolactin release induced by the acute administration of ethanol.
Assuntos
Etanol/farmacologia , Hipotálamo/efeitos dos fármacos , Adeno-Hipófise/efeitos dos fármacos , Substância P/metabolismo , Animais , Hipotálamo/química , Hormônio Luteinizante/sangue , Hormônio Luteinizante/metabolismo , Masculino , Adeno-Hipófise/química , Prolactina/sangue , Prolactina/metabolismo , Ratos , Ratos EndogâmicosRESUMO
Our data indicate that sex steroids modify the number of GABA receptors, as detected by a [3H]muscimol binding assay, in the tuberoinfundibular GABAergic system. GABA binding was affected by chronic hormonal treatments in different ways depending on the sex of the rats and the steroids administered. Estradiol increased GABA binding in ovariectomized female rats while testosterone decreased the number of GABA binding sites in gonadectomized male rats. These results suggest a sex difference in the regulation of hypothalamic GABA receptors.
Assuntos
Hormônios Esteroides Gonadais/farmacologia , Hipotálamo/metabolismo , Adeno-Hipófise/metabolismo , Receptores de GABA-A/metabolismo , Animais , Feminino , Hipotálamo/efeitos dos fármacos , Técnicas In Vitro , Masculino , Muscimol/metabolismo , Muscimol/farmacologia , Orquiectomia , Ovariectomia , Adeno-Hipófise/efeitos dos fármacos , Ratos , Ratos EndogâmicosRESUMO
A study was made of the effect of GABA-transaminase (GABA-T) inhibitors on the in vitro release of prolactin by pituitaries of male and female rats. Aminooxyacetic acid (AOAA) and gamma-acetylenic GABA (GAG) added to the incubation medium decreased prolactin release from both male and female rat pituitaries. Additive effects on prolactin release were only observed when male rat pituitaries were incubated with AOAA plus GABA. Prolactin concentration in the pituitary gland was also decreased by AOAA in both sexes. The present results lend support to the idea of an inhibitory action of GABA on prolactin release by the pituitary gland.
Assuntos
4-Aminobutirato Transaminase/antagonistas & inibidores , Prolactina/metabolismo , Alcinos , Aminocaproatos/farmacologia , Ácido Amino-Oxiacético/farmacologia , Animais , Feminino , Técnicas In Vitro , Masculino , Ovariectomia , Hipófise/metabolismo , Ratos , Ratos EndogâmicosRESUMO
The effects of Wallerian degeneration of the peripheral sympathetic neurons projecting to the hypothalamus on the mechanism of interaction between prolactin and substance P (SP) were examined. The effects of superior cervical ganglionectomy (SCGx) on SP content in various hypothalamic regions and in the hypophysis were evaluated in control and hyperprolactinemic rats. Male rats that received pituitary transplants at the age of 5 days and age-matched sham-operated controls were used. Pituitary grafting significantly increased circulating values of prolactin, as did SCGx. In hyperprolactinemic rats, SCGx partially decreased plasma prolactin levels. Neonatal hyperprolactinemia decreased SP content in the anterior (AH) and posterior (PH) hypothalamus and in the median eminence (ME), but increased it in the mediobasal hypothalamus (MBH). Acute SCGx significantly increased SP in the MBH, PH, and ME. SCGx in hyperprolactinemic animals further increased SP content in MBH. In the ME and Ah, SCGx in pituitary grafted rats decreased SP content as compared with the controls. In the pituitary gland (PG), SCGx only decreased SP content in hyperprolactinemic, but not in control rats. An interaction between peripheral nor-adrenergic neurons and prolactin to regulate SP within the hypothalamus was positive in the MBH, AH, ME, and PG, but not in the PH. These data indicate the existence of interactive mechanisms between prolactin and the peripheral sympathetic neurons to regulate SP content at the hypothalamic-pituitary axis. Interrelationships between prolactin and SP were also observed.
Assuntos
Hiperprolactinemia/metabolismo , Hipotálamo/metabolismo , Hipófise/metabolismo , Substância P/metabolismo , Sistema Nervoso Simpático/metabolismo , Degeneração Walleriana/metabolismo , Animais , Ganglionectomia , Hipotálamo Anterior/metabolismo , Hipotálamo Posterior/metabolismo , Masculino , Neurônios/metabolismo , Prolactina/sangue , Ratos , Ratos Wistar , Gânglio Cervical SuperiorRESUMO
The effect of a specific antiserum against vasoactive intestinal peptide (VIP) on GABA in the hypothalamic-pituitary axis was studied. The administration of anti-VIP serum (A-VIP) increased anterior pituitary GABA concentration in control rats, but decreased this neurotransmitter in rats with hyperprolactinemia induced by acute or chronic treatments with estrogens, or by the implanting of anterior pituitary glands under the kidney capsule. Besides, the injection of the A-VIP serum in the morning in proestrous rats causes a decrease in anterior pituitary GABA concentration, measured in the afternoon of the same day. The in vitro effect of A-VIP and VIP on endogenous GABA release from hypothalamic fragments and on anterior pituitary GABA concentration was studied. A-VIP increased both basal and high K(+)-evoked GABA effluxes whereas VIP produced a decrease in evoked GABA efflux from hypothalamic fragments. Furthermore, A-VIP inhibited the normal degradation of GABA that occurs in the isolated gland whereas VIP increased it. These results suggest that VIP modifies hypothalamic GABA release and anterior pituitary GABA concentration.
Assuntos
Sistema Hipotálamo-Hipofisário/fisiologia , Peptídeo Intestinal Vasoativo/fisiologia , Ácido gama-Aminobutírico/fisiologia , Animais , Implantes de Medicamento , Estradiol/farmacologia , Feminino , Sistema Hipotálamo-Hipofisário/efeitos dos fármacos , Hipotálamo/efeitos dos fármacos , Hipotálamo/fisiologia , Soros Imunes/administração & dosagem , Técnicas In Vitro , Masculino , Ovariectomia , Adeno-Hipófise/efeitos dos fármacos , Adeno-Hipófise/fisiologia , Proestro , Ratos , Ratos Endogâmicos , Peptídeo Intestinal Vasoativo/imunologia , Peptídeo Intestinal Vasoativo/farmacologia , Ácido gama-Aminobutírico/metabolismoRESUMO
In order to investigate the action of ethanol on the anterior pituitary gland, the effect of ethanol on prolactin secretion in vitro was studied. Ethanol significantly increased the in vitro incorporation of 3H-leucine into both prolactin contained within the pituitary gland and that released into the medium. The enhancement of 3H labelled-prolactin synthesis induced by ethanol was suppressed by cycloheximide. These results support the hypothesis that ethanol stimulates the in vitro synthesis and release of prolactin by the pituitary gland.
Assuntos
Etanol/farmacologia , Adeno-Hipófise/metabolismo , Prolactina/metabolismo , Acetaldeído/farmacologia , Animais , Cicloeximida/farmacologia , Técnicas In Vitro , Masculino , Adeno-Hipófise/efeitos dos fármacos , Radioimunoensaio , Ratos , Ratos EndogâmicosRESUMO
The 24h changes of glutamate (GLU) and aspartate (ASP) were studied in the median eminence (ME) and hypothalamic areas. It was analyzed whether prolactin may change their daily patterns. The hypothalamic concentration of these amino acids was measured by high-performance liquid chromatography (HPLC) with fluorometric detection. Plasma prolactin levels increased over the 24h light-dark cycle after pituitary grafting compared to controls, and its circadian rhythm was disrupted. In controls, aspartate and glutamate in the hypothalamic areas studied followed a specific daily variation or showed no rhythmicity. In the median eminence, hyperprolactinemia seem to phase advance the aspartate or glutamate peaks from 16:00 to 12:00. In the mediobasal hypothalamus, hyperprolactinemia altered daily changes of aspartate and significantly decreased its concentration. Also, it seems to delay the nocturnal glutamate peak compared to controls. In the posterior hypothalamus, hyperprolactinemia did not change aspartate and glutamate concentrations and their daily changes, although it increased the glutamine concentration. These data show the existence of 24h changes of amino acid concentration in three of the hypothalamic regions studied. Increased plasma prolactin levels differentially affected these patterns depending on the hypothalamic area analyzed.
Assuntos
Ácido Aspártico/metabolismo , Ritmo Circadiano/fisiologia , Ácido Glutâmico/metabolismo , Hiperprolactinemia/metabolismo , Hipotálamo/metabolismo , Eminência Mediana/metabolismo , Animais , Doença Crônica , Glutamina/metabolismo , Hipotálamo/anatomia & histologia , Masculino , Ratos , Ratos Wistar , Distribuição TecidualRESUMO
We studied the internalization and intracellular distribution of [3H] GABA in rat anterior pituitary cells. Electron microscopic autoradiography of anterior pituitary fragments or dispersed pituitary cells incubated with [3H] GABA showed that lactotrophs and, to a lesser extent, somatotrophs were the only cells that contained radioactive grains. Grain density analysis performed on dispersed pituitary cells after a pulse-chase experiment (10 min pulse and then change to a medium without radioactive GABA for various periods up to 2 h) revealed that GABA internalized by lactotrophs was distributed in various intracellular membranous organelles. Of the cell compartments examined, plasma membrane, Golgi apparatus, mitochondria and secretory granules had different time-dependent labeling patterns. The highest grain density values were associated with plasma membrane (at the first chase time) and the Golgi apparatus. Mitochondria and secretory granules also showed significant grain density values. A similar pattern of distribution was observed when fragments of prolactin-secreting pituitary adenomas were incubated with [3H] GABA. These results provide morphological data on the cellular specificity and intracellular distribution of GABA in anterior pituitary cells.