DRD1 基因敲除影响雄鼠生育的分子机理研究

摘要:目的 研究 DRD1 基因敲除引起雄鼠生殖功能下降的机制。 方法 本研究以 DRD1 基因敲除鼠( KO)和野生鼠( WT)为研究对象,对 8 周龄 KO 与 WT 雄鼠进行基因型鉴定,测体质量并计算睾丸系数,一侧睾丸 HE 染色观察形态,另一侧睾丸 TRIzol 法提取 RNA,通过 qRT-PCR 反应,采用相对定量法,分析与生殖密切相关基因的表达差异倍数。 结果 KO 与 WT 鼠体质量相比较具有显著差异( P<0. 05) 。 睾丸组织病理学切片观察发现,相对于 WT 鼠,KO 鼠生精小管排列疏松紊乱,生殖细胞层数少,提示精子形态及功能存在异常。 通过 qRT-PCR 进行基因表达差异分析,KO 小鼠相对 WT 小鼠 CFTR 表达上调 2. 32倍, Spata19 上调 4. 58 倍,SMCY 下调 5. 25 倍,DAZ 下调 3. 14倍。 结论 以上研究发现,敲除 DRD1 基因可通过影响睾丸组织结构及相关基因表达,进而影响生殖。

Study on Molecular Mechanism of Male Mice Reproduction in DRD1 Knockout

Abstract: Objective To study the mechanism of declining reproduction between DRD1 gene knockout mice ( KO) and wild mice ( WT) . Method Eight weeks male mice, including KO and WT mice, were weighted and the testis coefficient was calculating. The testis was observed by HE staining and reproduction-related genes were analyzed by qPCR. Result The result showed that KO and WT mice had significant difference in weight ( P <0. 05) . The HE staining showed that the seminiferous tubule of KO mice was derangement and germ cell had less cellular layer, which suggesting morphology and function of sperms may be abnormal in KO mice. For geneticexpression, CFTR was up-regulated 2. 32 fold, Spata19 was up-regulated 4. 58 fold, SMCY was down-regulated 5. 25 fold and DAZ was down-regulated 3. 14 fold between KO and WT mice, revealing that declining reproduction in KO male mice may related with the expression of reproduction-related genes. Conclusion In conclusion ,DRD1 gene could influence reproduction by genetic expression.