Abstract
The mechanisms by which GnIH regulates the steroid synthesis pathway in duck granulosa cells remain poorly understood. In this study, we measured steroid hormone secretion by ELISA and reproduction-associated gene expression by quantitative real-time Polymerase Chain Reaction (qPCR) in duck granulosa cells treated with different concentrations of GnIH (0, 0.1, 1, 10, and 100 ng/mL) for 24 h. The genome-wide expression profiles of GnIH-treated cells (0 and 10 ng/mL) were evaluated by high-throughput RNA sequencing. Compared with untreated cells, the secretion of the steroid hormones E2, E1, P4, and T was downregulated, with that of E1 and P4 reaching statistical significance (P<0.05); in contrast, the secretion of ACV and INH was significantly upregulated (P<0.05) after treatment with 10 and 100 ng/mL GnIH. The expression of encoding steroidogenic proteins and enzymes genes (STAR, CYP11A1, CYP17A1, CYP19A1, and 3-β-HSD) and encoding gonadotropin receptors genes (FSHR, LHR) were significantly declined (P<0.05) in the 10 and 100 ng/mL GnIH treatments. Transcriptome sequencing identified 348 differentially expressed genes (DEGs), including 253 upregulated and 95 downregulated genes. The DEGs were mainly involved in cell growth and death, immune response, and steroid biosynthesis pathways. We identified four novel DEGs (MROH5, LOC113840576, SDR42E1, and LOC113841457) with key roles in the regulation of steroid hormone biosynthesis. Our study revealed changes in gonadal steroid hormone secretion and steroid biosynthesis pathway-related gene expression in duck granulosa cells under the inhibitory effect of GnIH. These data contribute to our understanding of the molecular and genetic mechanisms underlying reproduction in ducks.
Highlights
In 2000, a hypothalamic neuropeptide isolated from the brain of Japanese quail, named gonadotropin inhibitory hormone (GnIH), was shown to inhibit the release of a gonadotropin (Tsutsui et al, 2000)
QPCR was performed to validate reproduction-associated gene expression in duck granulosa cells treated with different concentrations of GnIH (0, 0.1, 1, 10, and 100 ng/mL)
The expression of STAR, 3-β-HSD, CYP11A1, CYP17A1, CYP19A1, FSHR, LHR, and GnIHR was downregulated in the 0.1 and 1 ng/mL GnIH treatment groups compared with the control group (0 ng/mL GnIH) (Figure 2); only the expression of CYP11A1 reached statistical significance (P
Summary
In 2000, a hypothalamic neuropeptide isolated from the brain of Japanese quail, named gonadotropin inhibitory hormone (GnIH), was shown to inhibit the release of a gonadotropin (Tsutsui et al, 2000). GnIH plays an essential role in regulating animal reproduction and has inhibitory effects on reproductive regulation and gonadotropin expression. It can directly or indirectly inhibit the activity of gonadotropin-releasing hormone GnRH-expressing neurons in the hypothalamus to regulate the secretion and synthesis of pituitary gonadotropin (Ubuka et al, 2012).
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