Follicular fluid samples were obtained from the largest pre-ovulatory follicle of 120 women undergoing in-vitro fertilization and were examined for melatonin by enzyme-linked immunosorbent assay and the steroids oestradiol and progesterone by radioimmunoassay. The concentrations (mean +/- SE) of melatonin (213.4 +/- 18.9 pmol/l) and progesterone (20.1 +/- 1.1 mumol/l) in follicular fluid during the autumn and winter (dark) months were significantly higher than during the spring and summer (light) months, melatonin (138.4 +/- 12.5 pmol/l) and progesterone (11.6 +/- 0.8 mumol/l). By contrast, oestradiol concentrations were significantly lower during the dark months than during the light months (264.7 +/- 44.1 and 661.8 +/- 55.1 nmol/l respectively). There was a positive correlation between follicular fluid melatonin and progesterone concentrations (r = 0.271, P < 0.05, n = 120) and a negative relationship between melatonin and oestradiol (r = -0.254, P < 0.05, n = 120). The effects of melatonin alone and in combination with human chorionic gonadotrophin (HCG) or follicle stimulating hormone (FSH) on steroidogenesis by human granulosa cell culture were also investigated. Melatonin had minimal effects on oestradiol or progesterone production by granulosa cells. Interestingly, the oestradiol response in culture appeared to be different according to the time of the year when harvested. During the light period oestradiol production was enhanced. Melatonin also synergized with HCG in increasing progesterone production on days 6 and 7 after treatment during both light and dark periods. FSH stimulated oestradiol production by the cells on day 2 of culture. Melatonin had no effect on FSH stimulation of oestradiol production. The results of this study suggest that melatonin may be involved in the regulation of steroidogenesis by the human ovaries.
This study describes the hypophyseal angioarchitecture found in 79 adult New Zealand white rabbits. The pituitary glands and attached hypothalami were removed and carefully processed following routine histological methods, and the vascular organization was studied by light microscopy. Whole mounts of the pituitary median eminence complex were prepared and studied with a binocular dissecting microscope employing transmitted and epi-illumination. Arterial blood was found to be directed primarily to the neurohypophysis by the superior hypophyseal artery (SHA) and the inferior hypophyseal artery (IHA). A direct arterial blood supply was found to the adenohypophysis, but was limited solely to the pars intermedia by branches of the anterior hypophyseal artery (AHA) and the IHA. Capillaries of the pars intermedia were subdivided into an intermediate and a superficial plexus. The superficial plexus was situated between the intermediate plexus and the capillaries of the infundibular process. Capillaries of the superficial plexus did not form anastomoses between themselves, but ramified into the intermediate plexus to form a dense network of anastomosing capillaries that were continuous with capillaries of the pars distalis. A direct arterial blood supply was found only to the superficial plexus.
Melatonin levels in the blood of female rabbits were determined from the time of weaning to adulthood in a longitudinal study. Blood samples were taken at 3 h after light onset and 1 h after dark onset and plasma was analysed for melatonin, LH and FSH by radioimmunoassay procedures. The animals were sacrificed on days 25, 32, 39, 51, 72, 91 and 120 days of life and pituitaries removed for in vitro incubation of slices to determine their response to GnRH. Circulating melatonin levels were significantly higher in the dark compared to the light phase and peaked on days 72 and 91 when gonadotropin levels were at a nadir. Melatonin levels on day 120 were lower than those on any other day examined. Basal secretion of LH and FSH by pituitary slices in vitro increased several-fold from day 25 of age to days 51–91 and then decreased by day 120. The pituitary gonadotropin responsiveness to GnRH in vitro was also different: while LH generally increased, FSH accumulation remained constant after GnRH stimulation. These data suggest that the female rabbit pituitary undergoes changes in sensitivity to GnRH during sexual maturation and that the pineal gland may play a role in this process.
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