RESUMO
It is known that noradrenergic sympathetic nerve fibers connect the ovary and the spleen from the celiac ganglion. The modulation of the ovarian steroidogenesis in rats with polycystic ovary (PCO) by secretions of culture splenocytes from control (non PCO), PCO and PCO rats with superior ovarian nerve transection (PCO+SON-t) is investigated. Splenocytes from PCO rats increased progesterone (P) and decreasing estradiol (E) and androstenedione (A) release, a steroidogenic response different from that obtained with splenocytes of control rats. PCO also decreased the number of splenocyte beta-adrenergic receptors (betaR). SON transection reverted the effect of PCO on splenocytes betaR numbers and secretions of these splenocytes also reverted the stimulatory effect of PCO on P release, while norepinephrine (NE) treatment to PCO+SON-t splenocytes decreased their betaR number and their secretions restored the stimulation on progesterone release. Inversely, PCO+SON-t splenocyte secretions intensified the inhibition in estradiol with no effect on A. Treatment of PCO+SON-t splenocytes with NE or neuropeptide Y partially reverted the effects of PCO and SON-t The P and E-A response of PCO ovary might be differentially regulated by the splenocyte secretions through the neural connection involving ovary, SON, celiac ganglion and spleen and the neurotransmitter NE.
Assuntos
Neuroimunomodulação/fisiologia , Síndrome do Ovário Policístico/metabolismo , Esteroides/metabolismo , Animais , Feminino , Ovário/inervação , Ovário/metabolismo , Ratos , Receptores Adrenérgicos beta/metabolismo , Baço/metabolismoRESUMO
Most of the fibres that constitute the superior ovarian nerve (SON) originate at the neuronal bodies of the coeliac ganglion and innervate rat ovarian stroma cells. The purpose of this work was to study the part played by innervation on ovarian release of progesterone on day 15 and at the end of pregnancy in an integrated in vitro system known as the coeliac ganglion-SON-ovary system. We also investigated, in the same system, whether there is some kind of inter-relationship between the effect of adrenergic agents and LH on progesterone release on day 15 of pregnancy. The coeliac ganglion and the ovary were incubated in separate compartments, linked by the SON. The ovary was immersed in 2 ml buffer solution (ovarian compartment) and the coeliac ganglion was immersed in 2 ml of a different buffer solution (ganglion compartment). Under these conditions, the accumulation of progesterone in the ovarian compartment medium was used as an endpoint. Conditions were standardised on day 15 of pregnancy, when the decrease in the release of ovarian progesterone caused by non-specific stimulation on the ganglion with KCl (56 mM) demonstrated the functional integrity of the system. Neural influence was evaluated by the addition of adrenergic agents at a concentration of 10(-6)M to the coeliac ganglion. On day 15 of pregnancy, noradrenaline and propranolol increased progesterone release while phentolamine diminished it. The existence of ganglionic tone was assessed by analysing progesterone basal levels at different stages of pregnancy. The highest secretion of progesterone was found to take place on day 15, diminishing as pregnancy advanced. In addition, adrenergic neural participation was studied during the physiological luteolysis occurring at the end of pregnancy. Major findings were that noradrenaline increased ovarian accumulation of progesterone on day 19 and decreased it on day 20, while propranolol and phentolamine diminished progesterone release on both days. In additional studies, some neuroendocrine aspects were investigated at a peripheral level. The addition of LH only to the ovarian compartment did not affect progesterone secretion. However, when LH in the ovarian compartment was accompanied by noradrenaline, propranolol or phentolamine in the ganglion compartment, the release of progesterone decreased. It can be concluded that modifications of the neural state of the coeliac ganglion affect ovarian progesterone secretion and the physiology of pregnancy via the SON. The results may confirm that the coeliac ganglion-SON-ovary system provides a direct link between the autonomic nervous system and physiological events during pregnancy.
Assuntos
Adrenérgicos/farmacologia , Gânglios Simpáticos/metabolismo , Ovário/metabolismo , Prenhez/metabolismo , Progesterona/metabolismo , Agonistas Adrenérgicos beta/farmacologia , Animais , Técnicas de Cultura de Células , Feminino , Isoproterenol/farmacologia , Células Lúteas/efeitos dos fármacos , Células Lúteas/metabolismo , Hormônio Luteinizante/farmacologia , Ovário/efeitos dos fármacos , Ovário/inervação , Cloreto de Potássio/farmacologia , Gravidez , RatosRESUMO
This study investigates the interaction between the effect of epinephrine intracerebroventricular (icv) injection and LH on the progesterone concentration in ovarian vein blood (Po) in vivo, and also, on the release of ovarian progesterone and androstenedione in vitro, in rats on dioestrus day 2. When 2 mg ovine LH were injected in vein (i.v.), Po increased reaching 120+/-12.2 and 151+/-17.5 ng ml(-1) at 22 and 25 min, respectively. Another group of rats was injected intracerebroventricular with 5 microgram epinephrine at time zero, and with 2 mg ovine LH i.v. 3 min later. This time Po decreased during the first 3 min, then increased, reaching 64+/-7.1 ng ml(-1) at 25 min, lower than the Po obtained 22 min after LH i.v. injection only (P<0.01). Moreover, rats were injected i.v. with 2 mg ovine LH at time zero, and 7 min later with epinephrine intracerebroventricular. Po increased during the first 7 min, decreased until the 13th minute and reached 70+/-8.9 ng ml(-1) at 25 min, lower than the Po obtained 25 min after LH i.v. injection only (P<0.01). In other experience, rats with one (either right or left) superior ovarian nerve transected (SON-t), were injected intracerebroventricular with epinephrine. Five minutes later, the ovaries were removed and incubated in vitro with LH. Both ovaries (right or left) in which the SON was intact at time of epinephrine i. c.v. injection, showed a reduction of progesterone and androstenedione released in vitro (P<0.05). These results suggest that, on dioestrus day 2, the central adrenergic stimulus competes with LH in the release of ovarian progesterone. Also, the neural input that arrives at the ovary through the SON would antagonize the ovarian progesterone and androstenedione response to LH.
Assuntos
Androstenodiona/metabolismo , Epinefrina/administração & dosagem , Epinefrina/farmacologia , Hormônio Luteinizante/farmacologia , Ovário/irrigação sanguínea , Ovário/efeitos dos fármacos , Progesterona/metabolismo , Animais , Feminino , Injeções Intraventriculares , Ovário/inervação , Ovário/metabolismo , Ratos , Ratos Sprague-Dawley , Ovinos , Fatores de Tempo , Veias/efeitos dos fármacos , Veias/metabolismoRESUMO
The present study investigates the acute consequences of central adrenergic stimulation on the release of steroids from the ovary. The influence of the superior ovarian nerve (SON) and the relationship between the neural effect and peripheral LH levels were also examined. The intracerebroventricular (i.c.v.) injection of 5 microg epinephrine in SON-intact rats on day 1 of dioestrus (D1) increased progesterone levels in ovarian vein blood from 7 to 21 min after injection but the same injection in SON-intact rats on day 2 of dioestrus (D2) decreased progesterone levels in ovarian vein blood from 1 to 25 min. A smaller dose (0.5 microg) of epinephrine injected i.c.v. in SON-intact rats produced a decrease in progesterone levels in ovarian vein blood of shorter duration. In SON-transected (SONt) animals, 0.5 microg epinephrine i.c.v. caused a smaller decrease in progesterone levels compared with SON-intact rats (P<0.05). On the other hand, in SON-intact rats on D2, the i.c. v. injection of 0.5 microg epinephrine did not modify the peripheral LH levels during 25 min, but 5 microg epinephrine injected i.c.v. raised the peripheral LH level from the third minute after injection (P<0.05). Oestradiol levels in the ovarian vein blood did not change after epinephrine i.c.v. injection in rats on D2. To avoid any humoral influence, SONt and SON-intact rats on D2 were injected i.c.v. with 5 microg epinephrine or with vehicle, and 5 min later the ovaries were incubated in vitro with or without LH. Under these conditions, it was demonstrated that the previous injection of epinephrine in SON-intact rats resulted in a diminished release of progesterone from ovaries incubated with or without LH. These results suggest that a central adrenergic stimulus increases progesterone release from the ovary on D1 and decreases it on D2. Also, this neural input would arrive at the ovary through the SON, and would condition the ovarian response to LH on D2. Ovarian progesterone changes could be attributed to signals coming from ganglionar neurons, which are affected by the central adrenergic stimulation.
Assuntos
Glândulas Suprarrenais/efeitos dos fármacos , Epinefrina/farmacologia , Ovário/inervação , Progesterona/sangue , Animais , Ventrículos Cerebrais , Diestro/sangue , Estradiol/sangue , Feminino , Injeções Intraventriculares , Hormônio Luteinizante/sangue , Técnicas de Cultura de Órgãos , Ovário/irrigação sanguínea , Ovário/metabolismo , Progesterona/metabolismo , Ratos , Ratos Endogâmicos , Fluxo Sanguíneo Regional/efeitos dos fármacos , Estimulação Química , Fatores de TempoRESUMO
The aim of this study was to characterize the methylation system of phosphatidylethanolamine (PE) in the ovarian membranes and identify their possible role on the ovarian physiology. The presence of two methylating activities was found, by using S-adenosyl-L-[methyl-3H] methionine (SAM) as methyl donor. One of them (Met I) uses PE as substrate, and the other one, (Met II), catalyses the two successive methylations of phosphatidyl-N-monomethylethanolamine (PME) to phosphatidylcholine (PC). Met I shows a Km value of 5.71 microM and a Vm of 1.53 pmol/mg protein x min, working at pH 6.5, which seems to be the optimum. Met II exhibits low affinity for SAM, a Km of 50 microM, a Vm of 1.2 pmol/mg protein x min and it works an optimum pH of 9.5. Those enzymatic properties are in agreement with the amount of monomethylated and trimethylated products found working at pH 6.5 and 9.5, respectively. In order to investigate whether this enzyme system is affected or not by the hormonal environment influencing the ovary during the estrous cycle, the methylating activity was measured at its different stages. Both methylating activities were induced on proestrus but only Met I on diestrus. The total methylating activity increases when the ovarian membranes were obtained from rats injected with pregnant mare serum gonadotrophin (PMSG). Those results suggest that phospholipid methylation could be regulated by the hormonal environment during the estrous cycle.
Assuntos
Estro/metabolismo , Ovário/metabolismo , Fosfatidiletanolaminas/metabolismo , Animais , Estro/fisiologia , Feminino , Metilação , Fosfolipídeos/metabolismo , RatosRESUMO
The effect of androstenedione on luteal progesterone production was studied during luteolysis preceding parturition as well as that induced by the antiprogestin RU486 in late pregnant rats. Luteal cells from animals on days 19, 20 or 21 of pregnancy and incubated with 10 microM androstenedione increased progesterone production by 99, 136, and 277%, respectively. The animals receiving androstenedione (10 mg/rat s.c.) on day 19 of pregnancy showed an increase in serum progesterone levels, a decline in luteal 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) activity and an increase in corpus luteum weight without modifying 20 alpha-hydroxysteroid dehydrogenase (20 alpha-HSD) activity on day 21 of pregnancy. Androstenedione and testosterone but not dihydrotestosterone were able to prevent the decrease in serum progesterone concentration and corpus luteum weight observed 58 h after treatment with RU486 (2 mg/kg) on day 18 of pregnancy. However, the three androgens studied inhibited the luteal 3 beta-HSD activity but 20 alpha-HSD activity was not affected, when compared with animals receiving RU486 alone. The co-administration of androstenedione with the aromatase inhibitor 4-hydroxyandrostenedione or with the specific antioestrogen ICI 164,384 did not modify the effects induced by androstenedione in RU486-treated rats, indicating that the action of androstenedione on progesterone production and secretion at the time of luteolysis seems to occur through an androgenic mechanism and is not mediated by previous conversion of the androgens to oestrogens. In all experiments the high luteal 20 alpha-HSD activity, that characterizes a luteolytic process, was not modified by androgens. Androstenedione administered to adrenalectomized rats was also able to prevent the decrease in serum progesterone concentration observed in spontaneous or RU486-induced luteolysis. The administration of androstenedione to RU486-treated rats induced a decrease in luteal progesterone content concomitant with an increase in serum progesterone levels. These studies demonstrate that androgens during luteolysis, are able to stimulate luteal progesterone secretion, prevent the loss in corpora lutea weight and enhance the decrease in 3 beta-HSD activity, without affecting the increase in 20 alpha-HSD activity.
Assuntos
Androstenodiona/farmacologia , Corpo Lúteo/metabolismo , Luteólise/metabolismo , Mifepristona/farmacologia , Prenhez , Progesterona/biossíntese , 17-Hidroxiesteroide Desidrogenases/metabolismo , 20-Hidroxiesteroide Desidrogenases/metabolismo , 20-alfa-Hidroxiesteroide Desidrogenase , Adrenalectomia , Androstenodiona/análogos & derivados , Androstenodiona/metabolismo , Animais , Inibidores da Aromatase , Células Cultivadas , Corpo Lúteo/citologia , Corpo Lúteo/efeitos dos fármacos , Di-Hidrotestosterona/farmacologia , Estradiol/análogos & derivados , Estradiol/farmacologia , Feminino , Antagonistas de Hormônios/farmacologia , Luteolíticos/farmacologia , Indutores da Menstruação/farmacologia , Tamanho do Órgão/efeitos dos fármacos , Alcamidas Poli-Insaturadas , Gravidez , Progesterona/sangue , Ratos , Ratos Wistar , Testosterona/farmacologiaRESUMO
To determine if androstenedione, an aromatizable androgen, has a direct effect on luteal progesterone secretion, collagenase-dispersed luteal cells or whole corpora lutea from pregnant rats were incubated in the presence of the androgen. Luteal cells from 15-day pregnant rats responded to androstenedione in a dose-dependent manner, with an increase in progesterone output at doses of 1 and 10 microM, but with no effect at minor doses of the androgen. Luteal cells obtained from animals on day 4, 9, 15 or 19 of pregnancy and incubated with 10 microM of androstenedione, increased progesterone production by 243, 39, 84 and 146%, respectively. Androgens (androstenedione, testosterone or dihydrotestosterone) but no oestrogens (oestradiol or diethylstilboestrol) at a dose of 10 microM, stimulated progesterone production in incubated luteal cells obtained from 15-day pregnant rats. The time-course pattern of androstenedione-induced progesterone production was studied by superfusion experiments using corpora lutea from rats on day 15 of pregnancy. A significant progesterone output was observed when androstenedione, but not oestradiol, was perfused through the luteal tissue. Intrabursal ovarian administration of androstenedione (10 microM) to 19-day pregnant rats induced a significative increase in serum progesterone levels 8 and 24 h after treatment. These in vivo results confirm the stimulatory effect of androstendione on progesterone production obtained in incubated luteal cells from pregnant rats. This study reports a direct luteotrophic effect of androstenedione in rat corpus luteum, not mediated by previous conversion to oestrogens.