Wu Yu-Jie, Liu Yue, Hu Yan-Qin, Wang Li, Bai Fu-Rong, Xu Chen, Wu Jing-Wen
Shanghai Key Laboratory of Reproductive Medicine / Department of Histology, Embryology, Genetics and Developmental Biology, School of Medicine, Shanghai Jiao Tong University, Shanghai 200025, China .
Zhonghua Nan Ke Xue. 2020 Dec;26(12):1059-1067.
To investigate the role of miR-34b/c and miR-449 in maintaining the normal structure and function of efferent ductules and explore the molecular mechanism of infertility in miR-34b/c－/－ and miR-449－/－ dKO mice.
We observed the morphology of mouse efferent ductules by HE staining and analyzed the gene expressions in the efferent ductules of the wild-type and miR-34b/c－/－ and miR-449－/－ dKO mice by RNA sequencing. Then we screened the possible target genes of these two miRNA clusters and analyzed them along with the differentially expressed genes, followed by verification of the sequencing results by qRT-PCR.
Compared with the wild-type, the dKO mice showed morphologically abnormal efferent ductules and significantly decreased expressions of the genes involved in the formation of cilia and related to the transportation of water, ion and protein in the efferent ductules.
The deletion of miR-34b/c and miR-449 led to morphological abnormality of efferent ductules and dysfunction of aberrant cilia motility and reabsorption in the efferent ductules of dKO mice, resulting in infertility.
研究miR-34b/c和miR-449在维持输出小管正常结构和功能中的作用，并探讨miR-34b/c基因敲除（－/－）和miR-449基因敲除（－/－）双敲除（dKO）小鼠不育的分子机制。
通过苏木精-伊红（HE）染色观察小鼠输出小管的形态，并通过RNA测序分析野生型、miR-34b/c－/－和miR-449－/－ dKO小鼠输出小管中的基因表达。然后筛选这两个miRNA簇的可能靶基因，并与差异表达基因一起进行分析，随后通过实时定量逆转录聚合酶链反应（qRT-PCR）验证测序结果。
与野生型相比，dKO小鼠的输出小管形态异常，且参与纤毛形成以及与输出小管中水、离子和蛋白质运输相关的基因表达显著降低。
miR-34b/c和miR-449的缺失导致dKO小鼠输出小管形态异常，异常纤毛运动和重吸收功能障碍，从而导致不育。
