Idrees Muhammad, Kumar Vikas, Joo Myeong-Don, Ali Niaz, Lee Keun-Woo, Kong Il-Keun
Division of Applied Life Science (BK21 Four), Institute of Agriculture and Life Science (IALS), Gyeongsang National University, Jinju, South Korea.
Division of Applied Life Science, Department of Bio and Medical Big Data (BK21 Four), Research Institute of Natural Science (RINS), Gyeongsang National University (GNU), Jinju, South Korea.
Front Cell Dev Biol. 2021 Jan 22;8:611503. doi: 10.3389/fcell.2020.611503. eCollection 2020.
Src-homology-2-containing phosphotyrosine phosphatase (SHP2), a classic cytoplasmic protein and a major regulator of receptor tyrosine kinases and G protein-coupled receptors, plays a significant role in preimplantation embryo development. In this study, we deciphered the role of SHP2 in the somatic compartment of oocytes during meiotic maturation. SHP2 showed nuclear/cytoplasmic localization in bovine cumulus and human granulosa (COV434) cells. Follicle-stimulating hormone (FSH) treatment significantly enhanced cytoplasmic SHP2 localization, in contrast to the E treatment, which augmented nuclear localization. Enhanced cytoplasmic SHP2 was found to negatively regulate the expression of the ERα-transcribed and mRNAs, which are vital for oocyte meiotic arrest. The co-immunoprecipitation results revealed the presence of the SHP2/ERα complex in the germinal vesicle-stage cumulus-oocyte complexes, and this complex significantly decreased with the progression of meiotic maturation. The complex formation between ERα and SHP2 was also confirmed by using a series of computational modeling methods. To verify the correlation between SHP2 and /, SHP2 was knocked down RNA interference, and and mRNAs were analyzed in the control, E, and FSH-stimulated COV434 cells. Furthermore, phenyl hydrazonopyrazolone sulfonate 1, a site-directed inhibitor of active SHP2, showed no significant effect on the ERα-transcribed and mRNAs. Taken together, these findings support a novel nuclear/cytoplasmic role of SHP2 in oocyte meiotic resumption and maturation.
含Src同源2结构域的磷酸酪氨酸磷酸酶（SHP2）是一种经典的细胞质蛋白，也是受体酪氨酸激酶和G蛋白偶联受体的主要调节因子，在植入前胚胎发育中发挥重要作用。在本研究中，我们解析了SHP2在减数分裂成熟过程中卵母细胞体细胞区室中的作用。SHP2在牛卵丘细胞和人颗粒细胞（COV434）中呈现核/细胞质定位。与增强核定位的E处理相反，促卵泡激素（FSH）处理显著增强了细胞质SHP2的定位。发现增强的细胞质SHP2负向调节ERα转录的 和mRNA的表达，这对卵母细胞减数分裂阻滞至关重要。免疫共沉淀结果显示在生发泡期卵丘-卵母细胞复合体中存在SHP2/ERα复合物，并且随着减数分裂成熟的进行，该复合物显著减少。还通过一系列计算建模方法证实了ERα与SHP2之间的复合物形成。为了验证SHP2与 /之间的相关性，通过RNA干扰敲低SHP2，并在对照、E和FSH刺激的COV434细胞中分析 和mRNA。此外，活性SHP2的位点定向抑制剂苯基肼基吡唑酮磺酸盐1对ERα转录的 和mRNA没有显著影响。综上所述，这些发现支持SHP2在卵母细胞减数分裂恢复和成熟中具有新的核/细胞质作用。
