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

机译:猪不同发育阶段miR-34c及其靶基因的差异表达

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摘要

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.
机译:为了验证miR-34c的靶基因,使用了生物信息学软件来预测miR-34c的靶标。预测了miR-34c与精子发生和雄性生殖发育有关的三个可能的靶基因:锌指蛋白148(ZNF148),kruppel样因子4(KLF4)和血小板衍生的生长因子受体α(PDGFRA)。然后,通过定量聚合酶链反应检测miR-34c及其靶基因在猪睾丸组织不同发育阶段的表达。结果表明PDGFRA的表达与miR-34c负相关性最高。免疫组化染色观察2、3、4、5、5个月龄猪睾丸组织的形态,表明PDGFRA主要在睾丸早期发育阶段在基底膜附近的支持细胞中表达。组织,但PDGFRA的表达在后期逐渐降低。因此,蛋白质印迹分析表明PDGFRA的最高表达在2天大的睾丸组织中产生,并且在3和4个月大时表达水平降低,这与免疫组织化学染色的结果相关。总之,PDGFRA是miR-34c的靶基因。

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