西北农林科技大研究分析奶山羊精原细胞miRNA表达谱

摘要 : 西北农林科技大学的华进联教授和李新平教授领衔的课题组通过MACS法纯化了CD49f阳性和阴性的睾丸细胞，并利用Illumina测序技术分析了纯化后的CD49f阳性和阴性的细胞中的miRNA的表达谱。这项研究为奶山羊精原细胞提供了有用的miRNA表达谱数据，同时表明，CD49f可以用来富集奶山羊类精原细胞，包括SSCs。相关文章发表于2014年5月刊的《Journal of cellular biochemistry》杂志上。

micrornA(miRNA)在哺乳动物精子发生过程中发挥着重要的作用，精原细胞是精子发生的基础，对研究精子发生有重大价值。然而，奶山羊中的精原细胞miRNA的表达谱仍不清楚。来自西北农林科技大学的华进联教授和李新平教授领衔的课题组通过MACS法纯化了CD49f阳性和阴性的睾丸细胞，并对其中的miRNA表达谱进行了深入研究，其研究结果发表在5月刊的 Journal of cellular biochemistry上。

研究人员利用Illumina测序技术分析了纯化后的CD49f阳性和阴性的细胞中的miRNA的表达谱(小RNA测序由联川生物承担完成)，发现933个miRNAs在CD49​​f阳性细胞中的表达上调和916个miRNAs在CD49​​f阴性细胞中的表达上调分别两倍以上。

此外，在睾丸中多个特异于精原干细胞(SSCs)的miRNAs与标志基因在CD49​​f阳性睾丸细胞中有较高水平表达，包括miR-221，miR-23a，miR-29b，miR-24，miR-29a，miR-199b，miR-199a，miR-27a，miR-21和CD90，Gfra1，Plzf。对差异表达的miRNA进行生物信息学分析表明，这些miRNA在CD49​​f阳性细胞中的靶基因，参与了细胞周期的生物学过程和细胞周期的KEGG通路。

此次研究比较的miRNAome数据为奶山羊精原细胞提供了有用的miRNA表达谱数据，同时表明，CD49f可以用来富集奶山羊类精原细胞，包括SSCs。

原文摘要：

CD49f-Positive Testicular Cells in Saanen Dairy Goat Were Identified as Spermatogonia-Like Cells by miRNA Profiling Analysis

Jiang Wu, Mingzhi Liao, Haijing Zhu, Kai Kang, Hailong Mu, Wencong Song, Zhiwei Niu, Xin He, Chunling Bai, Guangpeng Li,Xinping Li andJinlian Hua

miRNAs, a type of small RNA, play critical roles in mammalian spermatogenesis. Spermatogonia are the foundation of spermatogenesis and are valuable for the study of spermatogenesis. However, the expression profiling of the miRNAs in spermatogonia of dairy goats remains unclear. CD49f has been one of the surface markers used for spermatogonia enrichment by magnetic activated cell sorting (MACS). Therefore, we used a CD49f microbead antibody to purify CD49f-positive and -negative cells of dairy goat testicular cells by MACS and then analysed the miRNA expression in these cells in depth using Illumina sequencing technology. The results of miRNA expression profiling in purified CD49f-positive and -negative testicular cells showed that 933 miRNAs were upregulated in CD49f-positive cells and 916 miRNAs were upregulated in CD49f-negative cells with a twofold increase, respectively; several miRNAs and marker genes specific for spermatogonial stem cells (SSCs) in testis had a higher expression level in CD49f-positive testicular cells, including miR-221, miR-23a, miR-29b, miR-24, miR-29a, miR-199b, miR-199a, miR-27a, and miR-21 and CD90, Gfra1, and Plzf. The BioInformatics analysis of differently expressed miRNAs indicated that the target genes of these miRNAs in CD49f-positive cells were involved in cell-cycle biological processes and the cell-cycle KEGG pathway. In conclusion, our comparative miRNAome data provide useful miRNA profiling data of dairy goat spermatogonia cells and suggest that CD49f could be used to enrich dairy goat spermatogonia-like cells, including SSCs. © 2014 Wiley Periodicals, Inc.

作者：联川生物

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