Zhou Linyan, Charkraborty Tapas, Yu Xiangguo, Wu Limin, Liu Gang, Mohapatra Sipra, Wang Deshou, Nagahama Yoshitaka
Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), School of Life Science, Southwest University, Chongqing, PR China.
BMC Dev Biol. 2012 Dec 7;12:36. doi: 10.1186/1471-213X-12-36.
In mammals, R-spondin (Rspo), an activator of the Wnt/β-catenin signaling pathway, has been shown to be involved in ovarian differentiation. However, the role of the Rspo/Wnt/β-catenin signaling pathway in fish gonads is still unknown.
In the present study, full-length cDNAs of Rspo1, 2 and 3 were cloned from the gonads of medaka (Oryzias latipes). The deduced amino acid sequences of mRspo1-3 were shown to have a similar structural organization. Phylogenetic analysis showed that Rspo1, 2 and 3 were specifically clustered into three distinct clads. Tissue distribution revealed that three Rspo genes were abundantly expressed in the brain and ovary. Real-time PCR analysis around hatching (S33-5dah) demonstrated that three Rspo genes were specifically enhanced in female gonads from S38. In situ hybridization (ISH) analysis demonstrated that three Rspo genes were expressed in the germ cell in ovary, but not in testis. Fluorescence multi-color ISH showed that Rspo1 was expressed in both somatic cells and germ cells at 10dah. Exposure to ethinylestradiol (EE2) in XY individuals for one week dramatically enhanced the expression of three Rspo genes both at 0dah and in adulthood.
These results suggest that the Rspo-activating signaling pathway is involved in the ovarian differentiation and maintenance in medaka.
在哺乳动物中,R-脊椎蛋白(Rspo)作为Wnt/β-连环蛋白信号通路的激活剂,已被证明参与卵巢分化。然而,Rspo/Wnt/β-连环蛋白信号通路在鱼类性腺中的作用仍不清楚。
在本研究中,从青鳉(Oryzias latipes)的性腺中克隆了Rspo1、2和3的全长cDNA。推导的mRspo1-3氨基酸序列显示具有相似的结构组织。系统发育分析表明,Rspo1、2和3分别特异性地聚集成三个不同的分支。组织分布显示,三个Rspo基因在脑和卵巢中大量表达。孵化前后(S33-5日龄)的实时PCR分析表明,从S38开始,三个Rspo基因在雌性性腺中特异性增强。原位杂交(ISH)分析表明,三个Rspo基因在卵巢的生殖细胞中表达,但在睾丸中不表达。荧光多色ISH显示,Rspo1在10日龄时在体细胞和生殖细胞中均有表达。XY个体暴露于乙炔雌二醇(EE2)一周后,在0日龄和成年期均显著增强了三个Rspo基因的表达。
这些结果表明,Rspo激活信号通路参与了青鳉卵巢的分化和维持。