| 摘要: |
| 【目的】研究TRIM8基因在小鼠组织和早期胚胎中的表达情况及其沉默后对早期胚胎体外发育的影响。【方法】采用RT-PCR、免疫印迹分别检测TRIM8基因及其编码的蛋白在小鼠不同组织（心、肝、脾、肺、肾、胃、肠、肌肉、子宫、卵巢和睾丸）中的表达情况，采用免疫组化技术对TRIM8在卵巢中进行定位。利用实时荧光定量PCR(RT-qPCR)、免疫印迹法检测TRIM8基因及其编码的蛋白在小鼠早期胚胎（1-细胞胚胎、2-细胞胚胎、4-细胞胚胎、8-细胞胚胎、桑葚胚和囊胚）中的表达规律，用免疫荧光结合激光共聚焦显微镜技术对TRIM8在早期胚胎中进行定位。利用RNA干扰技术沉默TRIM8基因，研究该基因沉默对小鼠早期胚胎体外发育的影响。【结果】TRIM8基因及其编码的蛋白在小鼠不同组织中均表达，但在子宫、卵巢和睾丸中表达水平相对较高。在卵巢中，TRIM8蛋白主要定位在不同发育阶段的卵母细胞中。TRIM8基因及其编码的蛋白在早期胚胎中均有表达，但其mRNA表达水平在1-细胞到4-细胞期胚胎中的表达水平显著高于8-细胞到囊胚期胚胎。TRIM8蛋白主要定位在不同时期胚胎的胞质中。TRIM8基因沉默组的囊胚率显著低于对照组(P<0.05)。【结论】TRIM8基因及其编码的蛋白在小鼠各组织及早期胚胎中均有表达，主要定位在早期胚胎的胞质中；沉默TRIM8基因会显著降低囊胚发育率。TRIM8可能参与调控小鼠早期胚胎的发育进程。 |
| 关键词: TRIM8基因 RNA干扰 早期胚胎发育 小鼠 |
| DOI： |
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| 基金项目:国家自然科学基金面上项目（31672415）；福建省自然科学基金面上项目（2018J01594） |
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| Expression and function of TRIM8 in tissues and early embyros of mouse |
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MA Zengyou,QIN Ge,ZHENG Haoyi,et al
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| Abstract: |
| 【Objective】In this study,the expression of tripartite motif 8 (TRIM8) gene in mouse tissues and early embryos and effects of TRIM 8 knockdown on in vitro development of early embryos were investigated.【Method】RT-PCR and western blot were used to assess the expression of TRIM8 gene and its encoded proteins in mouse tissues,and immunohistochemical technique was used to locate TRIM8 in ovary.The quantitative PCR and western blot were used to investigate the expression patterns of TRIM8TRIM 8 gene was knockdown through RNA interference technology to investigate its effect on vitro development of early embryos in mice.【Result】Expression of TRIM8 gene and its encoded proteins was detected in different tissues of mouse with high levels in uterus,ovary and testis.In ovary,TRIM8 protein was restricted to oocytes at various follicular stages.TRIM 8 gene and its encoded protein were expressed in early embryos with significantly higher mRNA expression level in 1-cell to 4-cell embryos than in periods from 8-cell to blastocyst embryos.TRIM8 protein was localized in the cytoplasm of embryos at different stages.The blastocyst rate of TRIM8 gene in knockdown group was significantly lower than that in the control group (P<0.05).【Conclusion】TRIM8 and its encoded protein were expressed in mouse oocytes and early embryos,mainly localised in the cytoplasm of early embryos.Knockdown of TRIM8 in zygotes sharply decreased the blastocyst rate.TRIM8 may be involved in regulating the development of early embryos in mice. |
| Key words: TRIM8 gene RNA interference early embryonic development mouse |
