YWHAZ在小鼠早期胚胎发育过程中的作用  

Function of YWHAZ in early embryonic development of mice

作　　者：赵怡凡 马增友 方俊博 孟碟方 彭辉[1] ZHAO Yifan;MA Zengyou;FANG Junbo;MENG Diefang;PENG Hui(College of Animal Science/College of Bee Science,Fujian Agriculture and Forestry University,Fuzhou 350002,China)

机构地区：[1]福建农林大学动物科学学院(蜂学学院),福建福州350002

出　　处：《中国兽医学报》2022年第3期541-546,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31672415)。

摘　　要：为了探究YWHAZ蛋白在小鼠早期胚胎发育过程中的表达、定位和作用,本试验利用RT-PCR和Western blot方法检测Ywhaz基因及其编码蛋白在小鼠早期胚胎发育中的表达;利用免疫荧光技术检测YWHAZ蛋白在小鼠早期胚胎发育过程中的定位;使用特异性的siRNA沉默合子中的Ywhaz,在胚胎发育的1.5,2.5,3.0,3.5,4.5,5.0 dpc分别观察2-细胞、4-细胞、8-细胞、桑葚胚和囊胚的发育率,阐明在合子中沉默Ywhaz基因对小鼠早期胚胎发育的影响。结果显示,Ywhaz mRNA及其编码蛋白在小鼠1-细胞、2-细胞、4-细胞、8-细胞、桑葚胚和囊胚中均有表达,YWHAZ蛋白定位在小鼠早期胚胎的细胞质和细胞核中。Ywhaz沉默组的2-细胞、4-细胞、8-细胞、桑葚胚和囊胚的发育率均极显著低于对照组(P<0.01),沉默组5.0 dpc囊胚率显著高于其4.5 dpc囊胚率(P<0.05)。结果表明,Ywhaz mRNA及其编码的蛋白在小鼠早期胚胎中有表达,其蛋白主要定位在早期胚胎的细胞核和细胞质中;在合子中沉默Ywhaz基因可导致小鼠早期胚胎发育率降低且存在明显的胚胎发育延迟现象。The aim of the present study was to analyse the expression, localization and function of YWHAZ protein during early embryonic development in mouse.RT-PCR and Western blot were used to detect the expression of Ywhaz mRNA and its encoded protein in mouse early embryonic development.The localization of YWHAZ protein in mouse early embryo was detected by immunofluorescence.The effects of Ywhaz knockdown using specific siRNA in zygotes on early embryonic development were elucidated through calculating the developmental rate of 2-cell, 4-cell, 8-cell stage embryo, morula and blastocyst at 1.5 dpc, 2.5 dpc, 3.0 dpc, 3.5 dpc, 4.5 dpc and 5.0 dpc, respectively.The results showed that Ywhaz mRNA and its encoded protein were expressed in 1-cell, 2-cell, 4-cell, 8-cell stage embryo, morula and blastocyst.Immunofluorescence result demonstrated that YWHAZ proteins were mainly localized in the cytoplasm and nucleus in preimplantation embryos.The developmental rate of 2-cell, 4-cell, 8-cell stage embryo, morula and blastocyst obtained from Ywhaz knockdown group was significantly reduced compared with the negative control(P<0.01).Meanwhile, the blastocyst rate of Ywhaz knockdown group at 5.0 dpc was significantly higher than its blastocyst rate at 4.5 dpc(P<0.05).The above results showed that Ywhaz mRNA and its encoded protein were expressed, and the protein was mainly localized in the nucleus and cytoplasm in mouse preimplantation embryos.Ywhaz knockdown in zygotes led to a lower development rate and a significant delay of early embryonic development in the mouse.

关 键 词：小鼠 YWHAZ蛋白 早期胚胎发育 

分 类 号：S852.1[农业科学—基础兽医学]