小鼠各期卵母细胞中SGK3 mRNA和蛋白的动态表达及其亚细胞定位  

Dynamic expressions and subcellular localization of SGK3 mRNA and protein in oocytes of mice at different stages

作　　者：何文宁 冯少青 庞海垚 孟峻[2] HE Wenning;FENG Shaoqing;PANG Haiyao;MENG Jun(Department of Clinical Laboratory Diagnostics,Affiliated Hospital,Inner Mongolia Medical University,Hohhot 010059,China;Department of Clinical Laboratory,Affiliated Hospital,Inner Mongolia Medical University,Hohhot 010050,China)

机构地区：[1]内蒙古医科大学临床检验诊断教研室,内蒙古呼和浩特010059 [2]内蒙古医科大学附属医院检验科,内蒙古呼和浩特010050

出　　处：《吉林大学学报（医学版）》2023年第5期1210-1216,共7页Journal of Jilin University：Medicine Edition

基　　金：内蒙古自治区科技厅自然科学基金项目(2021MS08158)。

摘　　要：目的:检测小鼠各期卵母细胞中血清和糖皮质激素诱导蛋白激酶3(SGK3)mRNA和蛋白的表达及其亚细胞定位,为阐明SGK3对小鼠卵母细胞周期变化的调控作用机制提供依据。方法:通过超排卵技术收集小鼠生发泡(GV)期、生发泡破裂(GVBD)期、第1次减数分裂(MⅠ)期卵母细胞和第2次减数分裂(MⅡ)期卵母细胞,采用实时荧光定量PCR(RT-qPCR)和Western blotting法检测各期小鼠卵母细胞中SGK3 mRNA和蛋白表达水平,收集GV、GVBD和MⅠ期卵母细胞后采用Western blotting法检测小鼠各期卵母细胞中Cdc25B-Ser30的磷酸化表达情况,免疫荧光法观察小鼠各期卵母细胞中SGK3亚细胞定位情况。结果:在小鼠各期卵母细胞中均有SGK3mRNA和蛋白表达。与GV期比较,GVBD和MⅡ期卵母细胞中SGK3 mRNA和蛋白表达水平升高(P<0.01);与GVBD期比较,MⅠ期卵母细胞中SGK3 mRNA和蛋白表达水平降低(P<0.01),MⅡ期卵母细胞中SGK3 mRNA和蛋白表达水平升高(P<0.01);与MⅠ期比较,MⅡ期卵母细胞中SGK3 mRNA和蛋白表达水平升高(P<0.01)。Cdc25B-Ser30在GV期被去磷酸化,在GVBD和MⅠ期被磷酸化。小鼠的GV期卵母细胞中SGK3定位于细胞核膜,在GVBD期前进入细胞核,在GVBD期定位于细胞核,MⅠ期则分布于整个细胞,在MⅡ期则由细胞浆再次进入细胞核。结论:小鼠各期卵母细胞中SGK3均呈动态表达,SGK3存在核浆穿梭,SGK3定位的动态变化可能参与了细胞周期B的激活。Objective:To discuss the expressions of serum and glucocorticoid-regulated kinase 3(SGK3)mRNA and protein in the oocytes of the mice at different stages and their subcellular localizations,and to provide the evidence for elucidating the regulatory mechanism of SGK3 in the cell cycle of oocytes of the mice.Methods:The oocytes of the mice at the germinal vesicle(GV)stage,germinal vesicle breakdown(GVBD)stage,first meiotic division(MⅠ)stage,and second meiotic division(MⅡ)stage were collected by using superovulation techniques.Real-time fluorescence quantitative PCR(RT-qPCR)and Western blotting methods were used to detect the expression levels of SGK3 mRNA and protein in the oocytes at different stages;the phosphorylation expressions of Cdc25B-Ser30 in the oocytes at different stages of the mice were detected by Western blotting method after collecting the oocytes at GV,GVBD,and MⅠstages;the subcellular localization of SGK3 in the oocytes of the mice at different stages was observed by immunofluorescence.Results:SGK3 mRNA and protein were expressed in the oocytes of the mice at all stages.Compared with GV stage,the expression levels of SGK3 mRNA and protein in the oocytes at GVBD and MⅡstages were increased(P<0.01).Compared with GVBD stage,the expression levels of SGK3 mRNA and protein in the oocytes at MⅠstage were decreased(P<0.01),and the expression levels of SGK3 mRNA and protein in the oocytes at MⅡstage were increased(P<0.01).Compared with MⅠstage,the expression levels of SGK3 mRNA and protein in the oocytes at MⅡstage were increased(P<0.01).The Cdc25B-Ser30 was dephosphorylated at GV stage and phosphorylated at GVBD and MⅠstages.The SGK3 was localized at the nuclear membrane of the oocytes at GV stage,and entered the nucleus before GVBD stage,and localized at the nucleus during GVBD stage,distributed throughout the oocytes during MⅠstage,and re-entered the nucleus from the cytoplasm during MⅡstage.Conclusion:SGK3 is dynamically expressed in the oocytes of the mice at different stages,and

关 键 词：血清和糖皮质激素诱导蛋白激酶 卵母细胞 减数分裂 动态表达 亚细胞定位 

分 类 号：Q132[生物学—普通生物学]