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J Dairy Sci ; 102(5): 4662-4673, 2019 May.
Article in English | MEDLINE | ID: mdl-30879805

ABSTRACT

Accumulating studies have suggested that microRNA play a part in regulating multiple cellular processes, such as cell proliferation, apoptosis, the cell cycle, and embryo development. This study explored the effects of miR-101-2 on donor cell physiological status and the development of Holstein cow somatic cell nuclear transfer (SCNT) embryos in vitro. Holstein cow bovine fetal fibroblasts (BFF) overexpressing miR-101-2 were used as donor cells to perform SCNT; then, cleavage rate, blastocyst rate, inner cell mass-to-trophectoderm ratio, and the expression of some development- and apoptosis-related genes in different groups were analyzed. The miR-101-2 suppressed the expression of inhibitor of growth protein 3 (ING3) at mRNA and protein levels, expedited cell proliferation, and decreased apoptosis in BFF, suggesting that ING3, a target gene of miR-101-2, is a potential player in this process. Moreover, by utilizing donor cells overexpressing miR-101-2, the development of bovine SCNT embryos in vitro was significantly enhanced; the apoptotic rate in SCNT blastocysts was reduced, and the inner cell mass-to-trophectoderm ratio and SOX2, POU5F1, and BCL2L1 expression significantly increased, whereas BAX and ING3 expression decreased. Collectively, these findings suggest that miR-101-2 promotes BFF proliferation and vitality, reduces their apoptosis, and improves the early development of SCNT embryos.


Subject(s)
Apoptosis/genetics , Cattle/genetics , Embryonic Development/genetics , Gene Expression Regulation/genetics , MicroRNAs/metabolism , Animals , Blastocyst/metabolism , Cattle/growth & development , Cell Line , Cell Proliferation/genetics , Female , Fibroblasts/metabolism , MicroRNAs/genetics , Nuclear Transfer Techniques/veterinary , RNA, Messenger/metabolism
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