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

doi:10.13481/j.1671-587X.20230514

吉林大学学报(医学版) ›› 2023, Vol. 49 ›› Issue (5): 1210-1216.doi: 10.13481/j.1671-587X.20230514

• 基础研究 • 上一篇

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

何文宁¹,冯少青¹,庞海垚¹,孟峻²()

^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

收稿日期:2022-10-23 出版日期:2023-09-28 发布日期:2023-10-26
通讯作者: 孟峻 E-mail:nmfrank@163.com
作者简介:何文宁（1997－），女，河南省周口市人，在读硕士研究生，主要从事临床生物化学和生殖分子生物学方面的研究。
基金资助:
内蒙古自治区科技厅自然科学基金项目(2021MS08158)

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

Wenning HE¹,Shaoqing FENG¹,Haiyao PANG¹,Jun MENG²()

^1.Department of Clinical Laboratory Diagnostics，Affiliated Hospital，Inner Mongolia Medical University，Hohhot 010059，China
^2.Department of Clinical Laboratory，Affiliated Hospital，Inner Mongolia Medical University，Hohhot 010050，China

Received:2022-10-23 Online:2023-09-28 Published:2023-10-26
Contact: Jun MENG E-mail:nmfrank@163.com

摘要/Abstract

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SGK3 mRNA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。

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

Abstract:

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 MⅠ stages； 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 MⅡ stages were increased （P<0.01）. Compared with GVBD stage， the expression levels of SGK3 mRNA and protein in the oocytes at MⅠ stage were decreased （P<0.01），and the expression levels of SGK3 mRNA and protein in the oocytes at MⅡ stage were increased （P<0.01）. Compared with MⅠ stage， the expression levels of SGK3 mRNA and protein in the oocytes at MⅡ stage were increased （P<0.01）.The Cdc25B-Ser30 was dephosphorylated at GV stage and phosphorylated at GVBD and MⅠ stages.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 MⅠ stage， and re-entered the nucleus from the cytoplasm during MⅡ stage. Conclusion SGK3 is dynamically expressed in the oocytes of the mice at different stages， and it exhibits the nuclear-cytoplasmic shuttling. The dynamic changes in the SGK3 localization may be involved in the activation of cell cycle progression.

Key words: Serum and glucocorticoid-induced protein kinase, Oocyte, Meiosis, Dynamic expression, Subcellular localization

中图分类号:

Q132

何文宁,冯少青,庞海垚,孟峻. 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[J]. 吉林大学学报(医学版), 2023, 49(5): 1210-1216.

Wenning HE,Shaoqing FENG,Haiyao PANG,Jun MENG. Dynamic expressions and subcellular localization of SGK3 mRNA and protein in oocytes of mice at different stages[J]. Journal of Jilin University(Medicine Edition), 2023, 49(5): 1210-1216.

图/表 3

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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

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