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miR-34c及其预测靶基因在猪不同发育阶段睾丸组织中的差异表达

Differential Expression of miR-34c and Its Predicted Target Genes in Testicular Tissue at Different Development Stages of Swine.

作者信息

Zhang Xiaojun, Zhao Wei, Li Chuanmin, Yu Haibin, Qiao YanYan, Li Aonan, Lu Chunyan, Zhao Zhihui, Sun Boxing

出版信息

Asian-Australas J Anim Sci. 2015 Nov;28(11):1532-6. doi: 10.5713/ajas.15.0047.

Abstract

To verified the target genes of miR-34c, bioinformatics software was used to predict the targets of miR-34c. Three possible target genes of miR-34c related to spermatogenesis and male reproductive development: zinc finger protein 148 (ZNF148), kruppel-like factor 4 (KLF4), and platelet-derived growth factor receptor alpha (PDGFRA) were predicted. Then, the expression of miR-34c and its target genes were detected in swine testicular tissue at different developmental stages by quantitative polymerase chain reaction. The results suggested that the expression of PDGFRA has the highest negative correlation with miR-34c. Then immunohistochemical staining was done to observe the morphology of swine testicular tissue at 2-days and 3, 4, 5-months of age, which indicated that PDGFRA was mainly expressed in the support cells near the basement membrane during the early development stages of testicular tissue, but that the expression of PDGFRA was gradually reduced in later stages. Therefore, western blot analyzed that the highest expression of PDGFRA was generated in 2-days old testicular tissues and the expression levels reduced at 3 and 4-months old, which correlated with the results of immunohistochemical staining. In conclusion, PDGFRA is a target gene of miR-34c.

摘要

为验证miR-34c的靶基因,使用生物信息学软件预测miR-34c的靶标。预测出与精子发生和雄性生殖发育相关的miR-34c的三个可能靶基因:锌指蛋白148(ZNF148)、 Kruppel样因子4(KLF4)和血小板衍生生长因子受体α(PDGFRA)。然后,通过定量聚合酶链反应检测不同发育阶段猪睾丸组织中miR-34c及其靶基因的表达。结果表明,PDGFRA的表达与miR-34c的负相关性最高。接着进行免疫组织化学染色,观察2日龄以及3、4、5月龄猪睾丸组织的形态,结果表明,在睾丸组织早期发育阶段,PDGFRA主要表达于靠近基底膜的支持细胞中,但在后期其表达逐渐降低。因此,蛋白质免疫印迹分析表明,PDGFRA在2日龄睾丸组织中的表达最高,在3和4月龄时表达水平降低,这与免疫组织化学染色结果相关。总之,PDGFRA是miR-34c的靶基因。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/79dd/4647091/ac3ddcfdc949/ajas-28-11-1532f1.jpg

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