College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
Gene. 2013 Jul 25;524(2):105-13. doi: 10.1016/j.gene.2013.04.054. Epub 2013 May 1.
Natriuretic peptides (NPs) are involved in maintaining cardiovascular and fluid homeostasis, regulating reproductive processes and bone growth, and other numerous functions. To better understand the role of NPs in goat (Capra hircus), in the present study, full-length cDNAs of goat Nppa (natriuretic peptide precursor A), Nppb (natriuretic peptide precursor B) and Nppc (natriuretic peptide precursor C), respectively encoding ANP, BNP and CNP, were cloned from adult goat heart and ovary. The putative prepropeptide ANP (prepro-ANP) and prepro-CNP share a high amino acid sequence identity with other species. Real-time PCR showed that Nppa, Nppb and Nppc were widely expressed in adult goat tissues. The mRNA expression of Nppa and Nppb in the heart was extremely higher compared with other tissues. Nppc mRNA expression in the lung and uterus was also higher than in other tissues. The expression of Nppa, Nppb and Nppc genes was examined at different ovarian follicle stages using RT-PCR. The mRNAs of Nppa and Nppb were detected in secondary follicles as well as in COCs (cumulus-oocyte-complexes) and granulosa cells of antral follicles. However, the mRNA expression of Nppc was observed throughout ovarian follicle development, and it was especially higher in granulosa cells of antral follicles. In vitro, stimulating goat granulosa cells with FSH led to an increase in the expression of Nppc by dose- and time-dependent manners and a rapid decline was induced by LH stimulation, but the expression of Nppa and Nppb did not change after FSH or LH treatment. These results suggest that Nppc is a gonadotropin-induced gene in granulosa cells of goat ovary and CNP may be involved in the regulation of ovarian follicle development and oocyte maturation.
利钠肽(NPs)参与维持心血管和体液平衡,调节生殖过程和骨骼生长等多种功能。为了更好地了解 NPs 在山羊(Capra hircus)中的作用,本研究从成年山羊心脏和卵巢中分别克隆了编码 ANP、BNP 和 CNP 的全长 cDNA,即 Nppa(利钠肽前体 A)、Nppb(利钠肽前体 B)和 Nppc(利钠肽前体 C)。推测的前肽 ANP(前肽-ANP)和前肽-CNP 与其他物种具有很高的氨基酸序列同一性。实时 PCR 显示 Nppa、Nppb 和 Nppc 在成年山羊组织中广泛表达。与其他组织相比,Nppa 和 Nppb 在心脏中的 mRNA 表达水平极高。Nppc 在肺和子宫中的 mRNA 表达水平也高于其他组织。使用 RT-PCR 检查了 Nppa、Nppb 和 Nppc 基因在不同卵巢卵泡阶段的表达。在次级卵泡以及 COC(卵丘-卵母细胞复合物)和腔前卵泡的颗粒细胞中检测到 Nppa 和 Nppb 的 mRNAs。然而,Nppc 的 mRNA 表达贯穿于卵巢卵泡发育过程,在腔前卵泡的颗粒细胞中表达水平更高。在体外,用 FSH 刺激山羊颗粒细胞导致 Nppc 的表达呈剂量和时间依赖性增加,并被 LH 刺激迅速下降,但 FSH 或 LH 处理后 Nppa 和 Nppb 的表达没有变化。这些结果表明,Nppc 是山羊卵巢颗粒细胞中促性腺激素诱导的基因,CNP 可能参与调节卵巢卵泡发育和卵母细胞成熟。