International Peace Maternity and Child Health Hospital, Shanghai Jiaotong University, Shanghai, China.
Stem Cells Dev. 2013 Jun 15;22(12):1839-50. doi: 10.1089/scd.2012.0653. Epub 2013 Mar 6.
Sperm abnormalities are one of the main factors responsible for male infertility; however, their pathogenesis remains unclear. The role of microRNAs in the development of sperm abnormalities in infertile men has not yet been investigated. Here, we used human induced pluripotent stem cells to investigate the influence of miR-122 expression on the differentiation of these cells into spermatozoa-like cells in vitro. After induction, mutant miR-122-transfected cells formed spermatozoa-like cells. Flow cytometry of DNA content revealed a significant increase in the haploid cell population in spermatozoa-like cells derived from mutant miR-122-transfected cells as compared to those derived from miR-122-transfected cells. During induction, TNP2 and protamine mRNA and protein levels were significantly higher in mutant miR-122-transfected cells than in miR-122-transfected cells. High-throughput isobaric tags for relative and absolute quantification were used to identify and quantify the different protein expression levels in miR-122- and mutant miR-122-transfected cells. Among all the proteins analyzed, the expression of lipoproteins, for example, APOB and APOA1, showed the most significant difference between the two groups. This study illustrates that miR-122 expression is associated with abnormal sperm development. MiR-122 may influence spermatozoa-like cells by suppressing TNP2 expression and inhibiting the expression of proteins associated with sperm development.
精子异常是男性不育的主要因素之一，但其发病机制尚不清楚。miRNA 在男性不育患者精子异常发育中的作用尚未得到研究。在这里，我们使用人诱导多能干细胞来研究 miR-122 表达对这些细胞体外向精子样细胞分化的影响。诱导后，突变 miR-122 转染的细胞形成精子样细胞。DNA 含量的流式细胞术显示，突变 miR-122 转染的细胞来源的精子样细胞中，单倍体细胞群显著增加。在诱导过程中，突变 miR-122 转染的细胞中的 TNP2 和鱼精蛋白 mRNA 和蛋白水平明显高于 miR-122 转染的细胞。使用高通量等重同位素标记相对和绝对定量技术来鉴定和量化 miR-122 和突变 miR-122 转染的细胞中的不同蛋白表达水平。在所有分析的蛋白中，脂蛋白的表达，例如 APOB 和 APOA1，在两组之间表现出最显著的差异。这项研究表明，miR-122 的表达与精子异常发育有关。miR-122 可能通过抑制 TNP2 的表达和抑制与精子发育相关的蛋白表达来影响精子样细胞。
