Guan Guijun, Sun Kaiqing, Zhang Xi, Zhao Xiaomiao, Li Mingyou, Yan Yan, Wang Yunzhi, Chen Jianbin, Yi Meisheng, Hong Yunhan
Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, Ministry of Education, College of Fisheries and Life Sciences, Shanghai Ocean University, Shanghai 201306, China.
Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, Ministry of Education, College of Fisheries and Life Sciences, Shanghai Ocean University, Shanghai 201306, China.
Mech Dev. 2017 Feb;143:53-61. doi: 10.1016/j.mod.2016.12.006. Epub 2017 Jan 16.
Gonadal soma-derived factor (gsdf) is reported to be a male initiator in medaka based on loss- and gain- of function via targeted disruption, or transgenic over-expression. However, little is known about how gsdf promotes undifferentiated gonad entry into male pathways or prevents entry into the female pathway. We utilized a visible folliculogenesis system with a reporter cassette of dual-color fluorescence expression to identify difference between oocyte development from wildtype and gsdf deficiency medaka. A red fluorescent protein (RFP) is driven by a major component of the synaptonemal complex (SYCP3) promoter which enables RFP expression solely in oocytes after the onset of meiosis, and a histone 2b-EGFP fused protein (H2BEGFP) under the control of an elongation factor (EF1α) promoter, wildly used as a mitotic reporter of cell cycle. This mitosis-meiosis visible switch revealed that early meiotic oocytes present in gsdf deficiency were more than those in wildtype ovaries, corresponding to the decrease of inhibin expression detected by real-time qPCR analysis, suggesting gsdf is tightly involved in the process of medaka oocyte development at early stage.
据报道，基于通过靶向破坏或转基因过表达实现的功能丧失和功能获得，性腺体细胞衍生因子（gsdf）是青鳉中的雄性启动因子。然而，关于gsdf如何促进未分化性腺进入雄性途径或阻止进入雌性途径，人们知之甚少。我们利用了一个带有双色荧光表达报告盒的可见卵泡发生系统，以鉴定野生型和gsdf缺陷型青鳉的卵母细胞发育之间的差异。红色荧光蛋白（RFP）由联会复合体（SYCP3）启动子的主要成分驱动，该启动子使RFP仅在减数分裂开始后的卵母细胞中表达，以及在伸长因子（EF1α）启动子控制下的组蛋白2b-EGFP融合蛋白（H2BEGFP），其广泛用作细胞周期的有丝分裂报告基因。这种有丝分裂-减数分裂可见开关表明，gsdf缺陷型中存在的早期减数分裂卵母细胞比野生型卵巢中的更多，这与实时定量PCR分析检测到的抑制素表达降低相对应，表明gsdf在青鳉早期卵母细胞发育过程中密切相关。
