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Roles of pituitary growth hormone (GH) in female reproduction are well established. Autocrine and/or paracrine actions of GH in the mammalian ovary have additionally been proposed, although whether the ovary is an extra-pituitary site of GH expression in the laying hen is uncertain. This possibility has therefore been assessed in the ovaries of Hy-Line hens before (between 10-16 weeks of age) and after (week 17) the onset of egg laying. Reverse transcription/polymerase chain reaction (RT-PCR) analysis has consistently detected a full-length (690 bp) pituitary GH cDNA in ovarian stroma from 10 weeks of age, although GH expression is far lower than that in the pituitary gland or hypothalamus. GH mRNA is also present in small (>1-4 mm diameter) follicles after their ontogenetic appearance at 14 weeks of age and in all other developing follicles after 16 weeks of age (>4-30 mm diameter). Immunoreactivity for GH is similarly present in the ovarian stroma from 10 weeks of age and in small (<4 mm diameter) and large (>4-30 mm) follicles from 14 and 16 weeks of age, respectively. The relative intensity of GH staining in the ovarian follicles is consistently greater in the granulosa cells than in the thecal cells and is comparable with that in the follicular epithelium. A 321-bp fragment of GH receptor (GHR) cDNA, coding for the intracellular domain of the receptor, has also been detected by RT-PCR in the ovary and is present in stromal tissue by 10 weeks of age, in small follicles (<4 mm diameter) by 14 weeks of age, and in larger follicles (>4-30 mm diameter) from 16 weeks. GHR immunoreactivity has similarly been detected, like GH, in the developing ovary and in all follicles and is more intense in granulosa cells than in the theca interna or externa. The expression and location of the GH gene therefore parallels that of the GHR gene during ovarian development in the laying hen, as does the appearance of GH and GHR immunoreactivity. These results support the possibility that GH has autocrine and/or paracrine actions in ovarian function prior to and after the onset of lay in hens.

In nonbroody birds, participation of prolactin in the reproductive functions is still unknown and its role in the local regulation of ovarian activity has had little attention. Therefore, the aim of the present study was to determine whether in the domestic hen prolactin influences in vitro steroid secretion by white and yellow chicken ovarian follicles. Small white (1-4 mm), medium white (4-6 mm), large white (6-8 mm) and 3 largest yellow preovulatory follicles (F3-F1; F3

11-ketotestosterone (OT) is a typical androgen of male teleost fish, but information on the question if it is involved in the feedback regulation of pituitary gonadotropin II (GTH-II) secretion is controversial. We have therefore studied the effects of OT on gonadotropin releasing-hormone (GnRH) stimulated GTH-II secretion in male African catfish Clarias gariepinus). In vivo experiments were carried out with intact and castrated fish. OT plasma levels were increased by implantation of silastic capsules containing 11-ketoandrostenedione (OA) which is converted to OT in both intact and castrated fish. When intact males received OA- or blank-capsules, treatment with salmon gonadotropin releasing-hormone analogue (Des-Gly(10)-D-Arg(6)-sGnRH-NEt; 0.2 μg sGnRHa/kg body weight) elevated the plasma GTH-11 levels in both groups. However, the levels were about 2 times higher in blank- than in OA-implanted fish. When castrated fish received either blank-or OA-capsules, sGnRHa treatment led to plasma GTH levels significantly higher than in sham-operated fish. However, there was no difference between the blank- or OA-implanted castrates, though OA implantation led to a restoration of OT plasma levels. This suggests that replacement ofOT is insufficient to reverse castration-induced effects. In vitro experiments were carried out with pituitary tissue fragments using a static culture system. The tissue remained sensitive to sGnRHa (5 × 10(-9)M) for 4 days after the beginning of incubation. Preincubation of pituitary tissue for 24 hours with 25 ng OT/ml medium (80 nM) completely abolished the stimulatory effect of sGnRHa on GTH-II secretion. Tritiated OT was not metabolized by pituitary tissue during 6 hours of incubation. We conclude that 11-ketotestosterone, a quantitatively prominent and non-aromatizeable circulating androgen participates, at least in part by direct action on the pituitary, in the negative feedback regulation of GnRH-stimulated GTH-II secretion in male African catfish.

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