摘要
旨在探究GPR50在小鼠卵母细胞体外成熟进程中的表达规律及亚细胞定位.通过卵母细胞体外培养技术、qPCR技术及免疫荧光技术测定GPR50在小鼠不同时期卵母细胞的转录水平及亚细胞定位.结果表明,所建立的qPCR方法敏感性高、特异性强、重复性好且检测效率高,可用于测定GPR50在小鼠卵母细胞中转录水平的表达;GPR50在不同阶段小鼠卵母细胞中均有表达,生发泡破裂(GVBD)后,GPR50的表达量显著高于GV期(P<0.05);随着卵母细胞的发育,其表达量不断增高,到MII期达到顶峰,极显著高于GV、GVBD及MI期(P<0.01).随着卵母细胞的发育,GPR50大量表达于细胞质和细胞膜,但在成熟卵母细胞中,GPR50主要集中分布于细胞膜.以上结果说明,GPR50在小鼠卵母细胞体外成熟中发挥重要的作用,为解析GPR50在哺乳动物卵母细胞体外成熟进程的分子机制奠定了基础.
In order to explore the expression pattern and subcellular localization of GPR50 in the in vitro maturation(IVM) of mouse oocytes,the transcriptional level and subcellular localization of GPR50 in mouse oocytes at different stages were determined by in vitro oocyte culture,qPCR and immunofluorescence technology.The results showed that the established qPCR method had high sensitivity,strong specificity,good reproducibility and high detection efficiency,which could be used to determine the expression of GPR50 transcription level in mouse oocytes.GPR50 was expressed in mouse oocytes at different development stages.After germinal vesicle breakdown(GVBD),the expression of GPR50 was significantly higher than that in GV stage(P0.05).With the development of oocytes,the expression of GPR50 increased continuously,and reached the peak in MII stage,which was significantly higher than that in GV,GVBD and MI stages(P0.01).With the development of oocytes,GPR50 was abundantly expressed in the cytoplasm and cell membrane,but in mature oocytes,GPR50 was mainly concentrated in the cell membrane.These results indicated that GPR50 played an important role in the IVM of mouse oocytes,which laid the foundation for analyzing the molecular mechanism of GPR50 in the IVM process of mammalian oocytes.
作者
王甜
黄显朋
朱洪阳
吉文汇
付伟
殷实
兰道亮
WANG Tian;HUANG Xian-peng;ZHU Hong-yang;JI Wen-hui;FU Wei;YIN Shi;LAN Dao-liang(School of Animal&Veterinary Sciences,Southwest Minzu University,Chengdu 610041,China;Ministry of Education Key Laboratory of Qinghai-Tibet Plateau Animal Genetic Resource and Utilization,Southwest Minzu University,Chengdu 610041,China)
出处
《西南民族大学学报（自然科学版）》
CAS
2023年第3期245-254,共10页
Journal of Southwest Minzu University(Natural Science Edition)
基金
国家重点研发计划(2021YFD1600200)
国家自然科学基金-面上项目(31872361)
西南民族大学研究生创新型科研项目(CX2021SZ58)。
关键词
GPR50
小鼠卵母细胞
体外成熟
转录水平表达
亚细胞定位
GPR50
mouse oocyte
in vitro maturation
transcriptional level expression
subcellular localization
