Zhang Huan, Jiang Xiaohua, Zhang Yuanwei, Xu Bo, Hua Juan, Ma Tieliang, Zheng Wei, Sun Rui, Shen Wei, Cooke Howard J, Hao Qiaomei, Qiao Jie, Shi Qinghua
Hefei National Laboratory for Physical Sciences at MicroscaleSchool of Life Sciences, University of Science and Technology of China, Hefei, Anhui 230027, People's Republic of ChinaLaboratory of Germ Cell BiologyKey Laboratory of Animal Reproduction and Germplasm Enhancement in Universities of Shandong, Qingdao Agricultural University, Qingdao, Shandong, ChinaMRC Human Genetics UnitInstitute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, UKDepartment of Obstetrics and GynecologyReproductive Medical Centre, Peking University Third Hospital, Beijing, China.
Hefei National Laboratory for Physical Sciences at MicroscaleSchool of Life Sciences, University of Science and Technology of China, Hefei, Anhui 230027, People's Republic of ChinaLaboratory of Germ Cell BiologyKey Laboratory of Animal Reproduction and Germplasm Enhancement in Universities of Shandong, Qingdao Agricultural University, Qingdao, Shandong, ChinaMRC Human Genetics UnitInstitute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, UKDepartment of Obstetrics and GynecologyReproductive Medical Centre, Peking University Third Hospital, Beijing, ChinaHefei National Laboratory for Physical Sciences at MicroscaleSchool of Life Sciences, University of Science and Technology of China, Hefei, Anhui 230027, People's Republic of ChinaLaboratory of Germ Cell BiologyKey Laboratory of Animal Reproduction and Germplasm Enhancement in Universities of Shandong, Qingdao Agricultural University, Qingdao, Shandong, ChinaMRC Human Genetics UnitInstitute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, UKDepartment of Obstetrics and GynecologyReproductive Medical Centre, Peking University Third Hospital, Beijing, China.
Reproduction. 2014 Jul;148(1):43-54. doi: 10.1530/REP-13-0508. Epub 2014 Mar 31.
In mammals, the primordial follicle pool, providing all oocytes available to a female throughout her reproductive life, is established perinatally. Dysregulation of primordial follicle assembly results in female reproductive diseases, such as premature ovarian insufficiency and infertility. Female mice lacking Dicer1 (Dicer), a gene required for biogenesis of microRNAs, show abnormal morphology of follicles and infertility. However, the contribution of individual microRNAs to primordial follicle assembly remains largely unknown. Here, we report that microRNA 376a (miR-376a) regulates primordial follicle assembly by modulating the expression of proliferating cell nuclear antigen (Pcna), a gene we previously reported to regulate primordial follicle assembly by regulating oocyte apoptosis in mouse ovaries. miR-376a was shown to be negatively correlated with Pcna mRNA expression in fetal and neonatal mouse ovaries and to directly bind to Pcna mRNA 3' untranslated region. Cultured 18.5 days postcoitum mouse ovaries transfected with miR-376a exhibited decreased Pcna expression both in protein and mRNA levels. Moreover, miR-376a overexpression significantly increased primordial follicles and reduced apoptosis of oocytes, which was very similar to those in ovaries co-transfected with miR-376a and siRNAs targeting Pcna. Taken together, our results demonstrate that miR-376a regulates primordial follicle assembly by modulating the expression of Pcna. To our knowledge, this is the first microRNA-target mRNA pair that has been reported to regulate mammalian primordial follicle assembly and further our understanding of the regulation of primordial follicle assembly.
在哺乳动物中，原始卵泡库在围产期建立，它为雌性提供整个生殖生命期内所有可用的卵母细胞。原始卵泡组装失调会导致女性生殖疾病，如卵巢早衰和不孕。缺乏Dicer1（Dicer）的雌性小鼠，Dicer1是一种微小RNA生物合成所需的基因，其卵泡形态异常且不孕。然而，单个微小RNA对原始卵泡组装的作用仍 largely未知。在此，我们报告微小RNA 376a（miR-376a）通过调节增殖细胞核抗原（Pcna）的表达来调控原始卵泡组装，Pcna是我们之前报道的在小鼠卵巢中通过调节卵母细胞凋亡来调控原始卵泡组装的基因。在胎儿和新生小鼠卵巢中，miR-376a与Pcna mRNA表达呈负相关，且能直接结合到Pcna mRNA的3'非翻译区。用miR-376a转染的妊娠18.5天小鼠卵巢在蛋白质和mRNA水平上均表现出Pcna表达降低。此外，miR-376a过表达显著增加了原始卵泡数量并减少了卵母细胞凋亡，这与同时转染miR-376a和靶向Pcna的小干扰RNA的卵巢情况非常相似。综上所述，我们的结果表明miR-376a通过调节Pcna的表达来调控原始卵泡组装。据我们所知，这是首次报道的调控哺乳动物原始卵泡组装的微小RNA-靶标mRNA对，进一步加深了我们对原始卵泡组装调控的理解。
