The present study was undertaken to examine in vivo the effect of growth hormone (GH) on progesterone and estradiol levels and on cell proliferation and apoptosis in the chicken ovary during sexual maturation. Hy-Line chickens (10 weeks old) were injected three times a week with 200 μg recombinant chicken GH (cGH) per kilogram body weight until sexual maturity. GH treatment significantly increased ovarian weight at 16 weeks of age, i.e., ~1 week before onset of egg laying. The progesterone content in the ovary just before and at the time of sexual maturity and the estradiol content before onset of egg laying were also elevated after cGH injections. The highest number of proliferating (positive for proliferating cell nuclear antigen) and apoptotic (positive for terminal-deoxynucleotidyl-transferase-mediated dUTP nick-end labeling) cells was found in the ovarian stroma and white follicles (>1-4 mm diameter), whereas the lowest number of these cells was detected in yellow (>8-30 mm) follicles. Administration of cGH significantly stimulated cell proliferation and inhibited cell apoptosis in the ovarian stroma and small ovarian follicles. The number of ovarian follicles and the weight of the ovary prior to the first oviposition were also higher in cGH-injected hens. Thus, prior to and after the onset of egg laying, GH participates in the growth, maturation and hormonal activity of ovarian follicles in the chicken, via the regulation of steroidogenesis, proliferation and apoptosis processes.

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.

Thirty-four-week-old laying hens received injections of recombinant chicken leptin to assess the role of leptin in avian ovarian function. In the first experiment, the hens (n = 60) were divided into three groups: (i) fed ad libitum; (ii) fasted; and (iii) fasted + leptin. Hens were fasted for 5 days and those treated with leptin received 250 g leptin kg

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