Wang S B, Liu Y P, Zhang Z H, Wang Z K, Xu Y X, Wang Z C
Laboratory of Sport Physiology and Biomedicine, School of Physical Education and Sport Sciences, Fujian Normal University, Fuzhou, China.
Provincial Key Laboratory for Developmental Biology and Neurosciences, College of Life Sciences, Fujian Normal University, Fuzhou, China.
Genet Mol Res. 2016 Jan 8;15(1):gmr7152. doi: 10.4238/gmr.15017152.
Frizzled 3 is an important receptor in the Wnt/β-catenin pathway, a conserved signaling pathway that regulates gene expression and controls diverse developmental processes. However, the role of this protein during follicular development in the adult ovary is not known. The present study was designed to investigate the expression and localization of Frizzled 3 mRNA and protein during the estrous cycle in the mouse ovary through in situ hybridization (ISH), real-time quantitative polymerase chain reaction, immunohistochemistry and western blot. ISH results showed that in proestrus, high expression of Frizzled 3 was found in the granulosa and stroma with weak levels in the corpus luteum. In estrus and diestrus, the stroma had high Frizzled 3 expression, but levels were low in granulosa cells and corpus luteum. In the metestrus, moderate expression of Frizzled 3 was found in the stroma but low to no expression was found in luteal cells and follicles. The mRNA and protein levels of Frizzled 3 were found to be the highest in proestrus and diestrus compared to estrus and metestrus (P < 0.05), confirming the ISH results. During estrus and diestrus, high Frizzled 3 expression was observed in the stroma and moderate levels in granulosa cells, and during estrus and proestrus, low expression was seen in the oocyte cell membrane. The western blot results further confirmed this change during the estrous cycle. Together, these results indicate that Frizzled 3 is involved in regulating follicular development and oocyte maturation during the estrous cycle.
卷曲蛋白3（Frizzled 3）是Wnt/β-连环蛋白信号通路中的一种重要受体，该信号通路是一条保守的信号通路，可调节基因表达并控制多种发育过程。然而，这种蛋白质在成年卵巢卵泡发育过程中的作用尚不清楚。本研究旨在通过原位杂交（ISH）、实时定量聚合酶链反应、免疫组织化学和蛋白质印迹法，研究卷曲蛋白3 mRNA和蛋白在小鼠卵巢发情周期中的表达及定位。ISH结果显示，在发情前期，颗粒细胞和基质中卷曲蛋白3高表达，黄体中表达较弱。在发情期和间情期，基质中卷曲蛋白3表达较高，但颗粒细胞和黄体中表达较低。在动情后期，基质中卷曲蛋白3中度表达，但黄体细胞和卵泡中低表达或无表达。与发情期和动情后期相比，发情前期和间情期卷曲蛋白3的mRNA和蛋白水平最高（P<0.05），证实了ISH结果。在发情期和间情期，基质中卷曲蛋白3高表达，颗粒细胞中中度表达，而在发情期和发情前期，卵母细胞膜上低表达。蛋白质印迹结果进一步证实了发情周期中的这种变化。这些结果共同表明，卷曲蛋白3参与发情周期中卵泡发育和卵母细胞成熟的调节。
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