Regulation of luteinizing hormone receptor in hippocampal neurons following different long-lasting treatments of castrated adult rats.

The aim of this study was to investigate the effects of different Luteinizing hormone (LH) and steroid hormones levels on LH receptor (LHR) expression in the hippocampal cells. Rats (24 males and 24 females) were assigned to four groups: one control and three experimental [gonadectomy (GDX), gonadectomy + gonadotropin releasing hormone analogue (GDX+GnRHa) and GDX+GnRHa+estradiol (E2) or testosterone (T)] independently for each gender. All experimental rats were gonadectomized; then GnRHa was administrated to GDX+GnRHa group, and GnRHa plus steroid hormone to GDX+GnRHa+E2 or T group in both genders for four-month. LHR mRNA expression and its protein level in hippocampal cells were measured using QRT-PCR and Western blotting. Quantification of mRNA revealed a decrease in LHR transcripts level in GDX+GnRHa group of females. A significant change was observed between GDX groups and GDX+GnRHa+E2 or T versus GDX+GnRHa group in females. High levels of LH decreased significantly the immature isoform of LHR in GDX group compared to control group in both genders, but low LH concentrations in GDX+GnRHa group induced immature LHR isoform production only in females. Therefore increased LH concentration induces production of incomplete LHR transcripts in hippocampal cells and decreases immature LHR at the protein level. This implies that LH decreases the efficiency of translation through either producing non-functional LHR molecules or preventing their translation.

Lactation inhibits the potentiating effect of galanin upon the GnRH-induced LH release observed in diestrous-1 rat

Recent demonstrations of no changes in hypothalamic gonadotropin releasing hormone (GnRH) gene expression nd GnRH levels detected at the pituitary gland in diestrous and lactating rats, indicate that lactational hypogonadotropism in this species is not associated with inhibition of hypothalamic GnRH synthesis and secretion. Hypothalamic galanin potentiates GnRH effects on luteinizing hormone (LH) secretion in male and cycling rats. To explore the interaction between GnRH and galanin during lactation, we studied in vitro the effects of pulsatile stimulation with those peptides upon LH synthesis and secretion from rat pituitaries on diestrous 1 or day 10 of lactation. Hemipituitaries were separately incubated in 1 ml Dulbecco's Minimal Essential Medium supplemented with 1 per cent penicillin-streptomycin and fetal calf serum, at 37 degrees Celsius in 5 per cent CO2-air. The hemipituitaries were stimulated during 12 h with hourly pulses, 6 min each, of (a) gonadotropin releasing hormone (GnRH 25 ng/pulse), (b) rat galanin (600 ng/pulse), (c) GnRH plus galanin, or (d) saline. Medium was collected before each pulse to determine LH by radioimmunoassay. After the 12 h pulsatile regime total RNA was extracted and both actin and beta-LH mRNA were determined by reverse transcriptase polymerase chain reaction. There was a significant stimulation of LH secretion by GnRH (ANOVA, p<0.001) without significant differences between diestrous and lactation pituitaries. Galanin alone did not modify LH secretion but it potentiated the effect of GnRH upon pituitaries from diestrous (p=0.036) but not lactating rats. Neither peptide alone or its combination modified pituitary beta-LH mRNA levels. Results show that galanin regulates differently the secretion and synthesis of LH at the pituitary level. The disappearance of galanin-induced potentiation of GnRH effects upon LH secretion during lactation might contribute to the hypogonadotropism of lactation in the rat.

Interacción de los péptidos opiodes endógenos con la función del eje hipotálamo-hipófisis-testículo / Interaction of endogenous opoid peptides with hypothalamie-hypophisis-testis axis function

A partir del aislamiento e identificación de sustancias endógenas con actividad opoide (endorfinas y encefalinas) (1), y del descubrimiento de los receptores opoides en el cerebro de mamíferos (2), numerososo estudios se han realizado con el fin de conocer el papel que estos opoides endógenos tienen en la regulación neuroendocrina del eje hipotálamo-hipófisis-gónada. Tanto los opioides endógenos como los agentes químicos con actividad opoide (morfina, heroína, metadona, análogo sintéticos de encefalinas, etc.) interfieren con la producción de las hormonas secretadas por la hipófisis; el efecto de estos opoides es inhibitorio para algunas hormonas como la LH, FSH y la TSH, mientras que para otras como la PRL, GH y ACTH estimulan su producción (3), los opoides actúan a nivel hipotalámico y suprahipotalámico modulando la secreción de factores liberadores e inhibitorios. En varones adultos sanos los péptidos opioides endógenos (POE) produce una disminución de los niveles séricos de gonadotrofinas. La administración de antagonistas específicos de los receptores opiáceos aumentan la liberación de hormona luteinizante (LH) e incrementan la freuencia y amplitud de los pulsos de LH (6). Tanto los efectos de los POE como de los antagonistas específicos de sus receptores están alterados en la diferentes patologías del eje hipotálamo-hipófisis-testículo

Melatonin: the assumed latest cure- all drug

This study was carried out to evaluate the hormonal and some biochemical changes which might be caused by administration of melatonin [Mt] daily for five months to 36 albino rats of both sexes. Long-term administration of doses equivalent to human dose range of melatonin caused an increase in testosterone, a decrease in estradiol [E2] levels in males with no change in the levels of both hormones in females, an increase in follicular stimulating hormones [FSH] and luteinizing hormone [LH] and a decrease in prolactin [PRL] levels in both sexes. Serum glutamicoxalo-acetic transaminase [GOT], glutamic- pyruvic transaminase [GPT] and sorbitol dehydrogenase [SDH] showed a significant increase from the third month onwards, while serum urea and creatinine showed a significant increase only at the fifth month

Maintenance of hypothalamic GnRH release during lactation in the rat: a push-pull perfusion study

The activity of the gonadotropin-releasing hormone (GnRH) pulse generator during lactation was assessed by direct determination of GnRH levels impinging upon the pituitary gland. Sprague-Dawley rats were implanted on day 15 of pregnancy with a push-pull perfusion cannula directed to the anterior pituitary. All implanted animals showed normal parturition, maternal behavior and lactation. Push-pull perfusions were performed in 15 rats suckling 11.0 +/- 0.8 pups (range 4-15) on day 7-20 of lactation and repeated on diestrous 1 after weaning in some of the same animals. GnRH content of the samples was assayed by RIA. GnRH pulses were clearly detected during lactation. Mean GnRH secretion rate was 1.9 +/- 0.3 pg/10 min (chi +/- SE, range between 0.5 and 3 pg/10 min) and interpulse interval was 37.5 +/- 1.7 min (range between 27 and 50 min). There was a significant decrease of about 19 percent in the interpulse interval after weaning. There was no significant difference in GnRH pulse amplitude nor in GnRH secretion rate between lactation and diestrous. These results demonstrate that nursing does not suppress the GnRH pulse generator in the rat

Estudio cinético de la pulsatilidad de la secreción de gonadotrofinas y testosterona en varones euspérmicos fértiles e infértiles con oligoastenozoospermia idiopática / kinetic study of pulsatility of gonadotropins and testosterone in euspermic males, fertile and infertile with idiopathic oligoasthenozoospermia

La oligoastenozoospermia idiopática es una de las causas de infertilidad masculina más frecuente y de etiopatogenia menos conocida. Para obtener información valiosa sobre la participación de algunos factores endocrinos en la etiología de este tipo de infertilidad, información que no es fácilmente obtenible mediante la utilización de métodos tradicionales, decidimos aplicar algunos algoritmos matemáticos recientemente propuestos para analizar con mayor exactitud la importancia de los pulsos de secreción de tres hormonas, la hormona luteinizante (LH), la hormona estimulante del folículo (FSH) y la testosterona (T). Se tomaron muestras de suero sanguíneo, cada 10 min. durante 12 h de 15 varones euspérmicos fértiles y 14 infértiles con oligoastenozoospermia idiopática y se analizaron los perfiles de concentración de FSH y T obtenidos mediante IRMA y RIA, así como las concentraciones de LH inmuno- y bioactiva determinadas mediante IRMA y bioensayo (JCEM 42:958, 1976). Con la finalidad de evaluar la reseva hipofisiaria de LH y FSH, depués de 8 h de secreción espontánea, se administraron (con 2 horas de diferencias) 2 pulsos intravenosos de 10 µg de un análogo de GnRH y se continuó el muestreo en la forma descrita, la pulsatilidad hormonal se analizó mediante un método computarizado de análisis de grupos de datos (Am J Phys 250:E486, 1986) y por el método de desconvolución de parámetros múltiples (JCEM 66:1291, 1988). En los varones infértiles se encontró una disminución significativa en la duración y frecuencia de los pulsos de LH comparados con los euspérmicos. Sin embargo, la vida media de LH, el intervalo interpulso, la amplitud y la masa secretada por pulso aumentaron en los pacientes infértiles comparados con los controles. El incremento de la vida media de la LH sugiere la secreción de una isoforma más ácida de esta hormona en el subgrupo infértil. Después de la inyección de GnRH la masa secretada y la concentración media de LH aumentaron significativamente en ambos grupos; este efecto fue mayor en los oligoastenozoospérmicos infértiles. En este gtupo se encontró también una disminución en el intervalo interpulso de LH bioactiva y por lo tanto más pulsoso durante el tiempo de muestreo, lo que produjo una mayor concentración de esta hormona en estos pacientes. Los varones oligoasternozoospérmicos secretaron aproximadamente 70 por ciento más LH bioactiva en respuesta a la primera inyección de GnRH que los controles normales. La desensibilización observada con LH inmunoactiva (disminución en la masa secretada después del segundo estímulo comparado con el primero) se observó también en el caso de la LH bioactiva. En los varones infértiles se observó una reducción significativa en la vida media de la FSH, comparada con los controles euspérmicos, hecho que sugiere que, al revés de lo que sucede en el caso de la LH, una isoforma más básica es secretada en estos pacientes. Además, la masa secretada por pulso de FSH es mayor, así como la tasa de secreción que alcanza valores 3 veces más altos en los varones infértiles y a pesar de la vida media menor de la hormona, su concentración media fue también mayor, lo que indica que la oligo-astenozoopermia se acompaña de sobresecreción de FSH en el estado basal; sin embargo, no se encontraron diferencias en la frecuencia de los pulsos de FSH. Después de las inyecciones de GnRH se observó un incremento de 6-7 veces en la masa de FSH secretada en ambos grupos estudiados. En todos los casos se secretó más FSH en el primer pulso después del GnRH, comparados con el segundo, lo que indica la desensibilización o agotamiento de las reservas de FSH. La testosterona en los varones oligoasternozoospérmicos presentó una reducción significativa en la frecuencia de los pulsos, pero las otras mediciones relacionadas con la secreción de testosterona no fueron diferentes entre los gurpos estudiados. Las diferencias observadas en los pacientes infértiles tanto en el patrón de secreción de FSH como de LH, inmuno- y bioactiva, reflejan alteraciones en el funcionamiento del eje hipotálamo-hipófisis que pueden influir en la deficiencia en la producción de espermatozoides, característica de este tipo de patología

Neuroendocrine mechanisms of lactational infertility in women

The current knowledge on the mechanisms of lactational infertility, discussed during a symposium of investigators in this subject, is reviewed. Three periods of lactation are examined: the first weeks postpartum, the period of extended lactational amenorrhea and the recovery of ovarian function. In the first postpartum weeks the inhibition of ovarian function is accounted by diminished pituitary response to GnRH, since exogenous GnRH fails to elicit a LH increase. Suckling can extend the period of ovarian inhibition for weeks, months or years, although it does not fully suppress pulsatile secretion of LH beyond the first weeks. Extended lactational amenorrhea is associated with low LH plasma levels, a great PRL increase in response to suckling, low basal E2 levels and a suppression of estrogen positive feedback. Decreased immunoreactive LH levels may result from partial suppression of the LH pulse generator and a smaller mass of GnRH released in each burst. The role of neurotransmitters, PRL and ovarian factors is discussed. After the recovery of ovulatory cycles suckling still has a residual infertility effect, associated to inadequate luteal function. The sources of variation among women and populations were recognized. Areas in which research is needed to improve the understanding of the mechanisms that sustain lactational amenorrhea are suggested

Effect of adenylate cyclase inhibitor and protein kinase C inhibitor on GnRH-induced LH release and LH beta subunit biosynthesis in rat anterior pituitary cells

According to our previous studies together with others, GnRH, a hypothalamic decapeptide, has been known to be a major regulator for LH release and its subunit biosynthesis in anterior pituitary gonadotropes. But the precise mechanisms by which GnRH exerts stimulatory effects on LH release and its subunit biosynthesis have not been clearly understood. In the present study we examined the effect of GnRH on protein kinase C (PKC) activity and intracellular cAMP content in cultured anterior pituitary cells of rat to clarify whether PKC or cAMP are involved in GnRH action. Moreover, we examined the effects of staurosporine (ST), a PKC inhibitor and 2',3'-dideoxyadenosine (2',3'-DDA), an adenylate cyclase inhibitor, on LH release and steady state LH beta subunit mRNA levels in cultured anterior pituitary cells of rat. PKC activity was rapidly increased within 30 min after GnRH treatment whereas intracellular cAMP level was elevated 18 h after GnRH treatment. ST significantly inhibited GnRH-induced LH release and LH beta subunit mRNA levels in a dose-dependent manner, showing an half maximal response at 50 nM ST. 2',3'-DDA inhibited GnRH-induced LH release and LH beta subunit mRNA levels in a dose-dependent manner in pituitary cells. From these results, it is suggested that GnRH stimulates LH beta subunit mRNA level as well as LH release in anterior pituitary cells and this GnRH action might be mediated by PKC activation and cAMP stimulation.

Effect of cimetidine on adenohypophysis of male albino mice under certain experimental condition.

Effect of 75 mg/kg of body weight of cimetidine administered intraperitoneally daily for 14 days to two groups of experimental animals (one of the experimental group was having intact testis and another group was bilaterally orchidectomized) was observed on cell population & cell volume of gonadotrophs and lactotrophs in pituitary gland, as it has not been studied earlier. In the experimental group of intact testis, there was significant reduction in the cell population of FSH cells in the cephalomedian area (P < 0.001) and in the lateral lobe (P < 0.01); the volume of both FSH and LH cells was also significantly reduced. In group 4 and group 5 there was significant increase in the population of lactotrophs and also in the volume of LH cells, FSH cells & lactotrophs. The change in the gonadotrophs in group 2 was due to increased production of testosterone from hypertrophied Leydig cells of testis rather then it's direct effect on adenohypophysis; in group 4 and group 5 the changes were due to lack of testosterone as in those cases bilaterally orchidectomy already done.