吉林大学学报(医学版) ›› 2023, Vol. 49 ›› Issue (4): 884-889.doi: 10.13481/j.1671-587X.20230408

• 基础研究 • 上一篇    下一篇

血清和糖皮质激素诱导蛋白激酶3在小鼠不同细胞周期1-细胞期受精卵中的表达水平及其亚细胞定位

庞海垚1,侯力1,何文宁1,孟峻2()   

  1. 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:2022-12-02 出版日期:2023-07-28 发布日期:2023-07-26
  • 通讯作者: 孟峻 E-mail:nmfrank@163.com
  • 作者简介:庞海垚(1999-),男,内蒙古自治区包头市人,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:
    国家自然科学基金项目(81360109);内蒙古自治区科技厅自然科学基金项目(2021MS08158)

Expression levels of serum and glucocorticoid-induced protein kinase 3 in one-cell stage fertilized eggs at different cell cycles in mice and its subcellular localization

Haiyao PANG1,Li HOU1,Wenning HE1,Jun MENG2()   

  1. 1.Department of Clinical Laboratory Diagnosis,Inner Mongolia Medical University,Hohhot 010059,China
    2.Department of Clinical Laboratory,Affiliated Hospital,Inner Mongolia Medical University,Hohhot 010050,China
  • Received:2022-12-02 Online:2023-07-28 Published:2023-07-26
  • Contact: Jun MENG E-mail:nmfrank@163.com

摘要:

目的 检测血清和糖皮质激素诱导蛋白激酶3(SGK3)在小鼠不同细胞周期1-细胞期受精卵中的表达水平及其亚细胞定位,初步阐明SGK3对哺乳动物受精卵早期发育的调控机制。 方法 通过超排卵技术和受精卵体外培养收集小鼠1-细胞期受精卵,分别采用实时荧光定量PCR法 (RT-qPCR)和Western blotting法检测小鼠不同细胞周期1-细胞期受精卵中SGK3 mRNA和蛋白表达水平,采用免疫荧光法观察SGK3在小鼠不同细胞周期1-细胞期受精卵中的亚细胞定位情况。 结果 在小鼠不同细胞周期1-细胞期受精卵中均有SGK3 mRNA和蛋白表达;与G1期、S期和M期比较,G2期小鼠1-细胞期受精卵中SGK3 mRNA和蛋白表达水平均明显升高 (P<0.01)。免疫荧光法检测,在G1期和S期SGK3(红色荧光)定位于小鼠1-细胞期受精卵细胞质,在G2期部分SGK3进入细胞核,在M期SGK3广泛分布于小鼠1-细胞期受精卵中。 结论 SGK3在小鼠不同细胞周期1-细胞期受精卵中动态表达,SGK3在受精卵细胞分裂过程中存在核浆穿梭,SGK3的定位改变可能推动小鼠1-细胞期受精卵G2/M转换和有丝分裂进程。

关键词: 血清和糖皮质激素诱导蛋白激酶3, 有丝分裂, 细胞周期, 受精卵

Abstract:

Objective To detect the expression levels of serum and glucocorticoid-induced protein kinase 3 (SGK3) in one-cell stage fertilized eggs at different cell cycles of the mice and its subcellular localization, and to preliminarily clarify the regulatory effect of SGK3 on the early development of fertilized eggs of the mammal. Methods The one-cell stage fertilized eggs of the mice were collected by superovulation technique and in vitro culture of fertilized eggs, the expression levels of SGK3 mRNA and protein in the one-cell stage fertilized eggs at different cell cycles were detected by real-time fluorescence quantitative PCR (RT-qPCR) and Western blotting methods, respectively; the subcellular localization of SGK3 in the one-cell stage fertilized eggs at different cell cycles of the mice was observed by immunofluorescence assay. Results The SGK3 mRNA and protein were expressed in the one-cell stage fertilized eggs at different cell cycles of the mice;compared with G1, S and M phases, the expression levels of SGK3 mRNA and protein in one-cell phase fertilized eggs at G2 phase were increased(P<0.01). The immunofluorescence results showed that SGK3 (red fluorescence) was localized in cytoplasm of the one-cell phase fertilized eggs at G1 and S phases, and some SGK3 entered into the nucleus at G2 phase,and the SGK3 was widely distributed throughout the cells at M phase. Conclusion The SGK3 in the one-cell stage fertilized eggs at different cell cycles of the mice dynamically expresses, and SGK3 is shuttered between the nucleus and cytoplasm during the process of fertilized egg cell division, and the altered localization of SGK3 may drive the G2/M transition and mitotic progression in one-cell stage fertilized eggs of the mice.

Key words: Serum and glucocorticoid induced protein kinase 3, Mitosis, Cell cycle, Fertilized eggs

中图分类号: 

  • Q132