Yu Jianning, Yan Leyan, Chen Zhe, Li Hui, Zhu Huanxi, Chen Rong, Shi ZhenDan
Key Laboratory of Protected Agriculture Engineering in the Middle and Lower Reaches of Yangtze River, Ministry of Agriculture; Laboratory of Animal Breeding and Reproduction, Institute of Animal Science, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China.
J Reprod Dev. 2018 Aug 20;64(4):343-350. doi: 10.1262/jrd.2018-001. Epub 2018 Jun 8.
Treatment of fetal rat and embryonic chicken with exogenous glucocorticoids induces premature differentiation of growth hormone (GH) secreting cells. The effect of corticosterone (CORT) on somatotroph differentiation was mostly studied in pituitary cells in vitro. Currently, there is no evidence for glucocorticoid-mediated induction of somatotroph differentiation during pituitary development in bird species other than chicken. In this study, we sought to find out if in ovo injection of corticosterone into developing goose embryos could induce premature increase of GH in somatotrophs. On embryonic day (e) 15, the albumen of fertile goose eggs was injected with 300 μl of 0.9% saline, 300 μl 5 × 10M CORT, or 300 μl 5 × 10 M CORT. Embryos were allowed to develop until e20 and e28 and isolated pituitaries were subjected to quantitative real-time PCR and immunocytochemistry to detect GH mRNA and protein, respectively. At e20 and e28, blood from chorioallantoic vessels was subjected to radioimmunoassay for analysis of plasma GH protein. In ovo administration of exogenous corticosterone brought about a 2.5-fold increase in the expression of GH mRNA and increased the in situ expression of GH protein in goose pituitary cells, and enhanced plasma GH levels compared to that of the respective controls at e20. These findings prove that administration of glucocorticoid could stimulate the expression of GH in somatotrophs during goose embryonic development. Our results suggest the probable involvement of membrane glucocorticoid receptor in the corticosterone mediated expression of GH. Together with previously published data, our results suggest that corticosterone mediated induction of GH expression during embryonic development is relatively conserved among different vertebrates.
用外源性糖皮质激素处理胎鼠和鸡胚可诱导生长激素(GH)分泌细胞过早分化。皮质酮(CORT)对生长激素细胞分化的影响大多是在体外垂体细胞中进行研究的。目前,除鸡以外,尚无证据表明糖皮质激素在鸟类垂体发育过程中介导生长激素细胞分化。在本研究中,我们试图探究向发育中的鹅胚卵内注射皮质酮是否会诱导生长激素细胞中GH过早增加。在胚胎第15天,向受精鹅蛋的蛋清中注射300 μl 0.9%生理盐水、300 μl 5×10⁻⁶ M CORT或300 μl 5×10⁻⁸ M CORT。胚胎继续发育至第20天和第28天,分离出的垂体分别进行定量实时PCR和免疫细胞化学检测,以检测GH mRNA和蛋白质。在第20天和第28天,采集尿囊血管中的血液进行放射免疫分析,以检测血浆GH蛋白。与相应对照组相比,卵内给予外源性皮质酮使鹅垂体细胞中GH mRNA表达增加2.5倍,GH蛋白的原位表达增加,并在第20天提高了血浆GH水平。这些发现证明,糖皮质激素给药可在鹅胚胎发育过程中刺激生长激素细胞中GH的表达。我们的结果表明,膜糖皮质激素受体可能参与了皮质酮介导的GH表达。结合先前发表的数据,我们的结果表明,皮质酮在胚胎发育过程中介导的GH表达在不同脊椎动物中相对保守。