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

  • Zhang, Xiaojun (College of Animal Science, Jilin University) ;
  • Zhao, Wei (College of Animal Science, Jilin University) ;
  • Li, Chuanmin (College of Animal Science, Jilin University) ;
  • Yu, Haibin (College of Animal Science, Jilin University) ;
  • Qiao, YanYan (College of Animal Science, Jilin University) ;
  • Li, Aonan (College of Animal Science, Jilin University) ;
  • Lu, Chunyan (College of Animal Science, Jilin University) ;
  • Zhao, Zhihui (College of Animal Science, Jilin University) ;
  • Sun, Boxing (College of Animal Science, Jilin University)
  • Received : 2015.01.16
  • Accepted : 2015.04.29
  • Published : 2015.11.01

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.

Keywords

miR-34c;PDGFRA;Testicular Tissue;Differential Expression

References

  1. Bouhallier, F., N. Allioli, F. Lavial, F. Chalmel, M. H. Perrard, P. Durand, J. Samarut, B. Pain, and J. P. Rouault. 2010. Role of miR-34c microRNA in the late steps of spermatogenesis. RNA 16:720-731. https://doi.org/10.1261/rna.1963810
  2. Chang, R. C., W. Ying, F. W. Bazer, and B. Zhou. 2014. MicroRNAs control macrophage formation and activation: The inflammatory link between obesity and cardiovascular diseases. Cells 3:702-712. https://doi.org/10.3390/cells3030702
  3. Geremia, R., C. Boitani, M. Conti, and V. Monesi. 1977. RNA synthesis in spermatocytes and spermatids and preservation of meiotic RNA during spermiogenesis in the mouse. Cell Differ. 5:343-355. https://doi.org/10.1016/0045-6039(77)90072-0
  4. Lee, Y., M. Kim, J. Han, K. H. Yeom, S. Lee, S. H. Baek, and V. N. Kim. 2004. MicroRNA genes are transcribed by RNA polymerase II. EMBO 23:4051-4060. https://doi.org/10.1038/sj.emboj.7600385
  5. Lian, C., B. Sun, S. Niu, R. Yang, and Z. Zhao. 2012 A comparative profile of the microRNA transcriptome in immature and mature porcine testes using Solexa deep sequencing. FEBS J. 279:964-975. https://doi.org/10.1111/j.1742-4658.2012.08480.x
  6. Mello, T. R., A. C. Aleixo, D. G. Pinheiro, F. M. Nunes, M. M. Bitondi, K. Hartfelder, A. R. Barchuk, and Z. L. Simoes. 2014. Developmental regulation of ecdysone receptor (EcR) and EcR-controlled gene expression during pharate-adult development of honeybees (Apis mellifera). Front Genet 5:445.
  7. Ning, Z., H. Zhu, F. Li, Q. Liu, G. Lig, T. Tan, B. Zhang, S, Chen, G. Li, D. Huang, S. J. Meltzer, and H. Zhang. 2014. Tumor suppression by miR-31 in esophageal carcinoma is p21-dependent. Genes Cancer 5:436-440.
  8. Reinhart, B. J., F. J. Slack, M. Basson, A. E. Pasquinelli, J. C. Bettinger, A. E. Rougvie, H. R. Horvitz, and G. Ruvkun. 2000. The 21-nucleotide let-7RNA regulates developmental timing in Caenorhabditis elegans. Nature 403:901-906. https://doi.org/10.1038/35002607
  9. Ren, J., H. Tang, X. Yan, X. Huang, B. Zhang, H. Ji, B. Yang, D. Milan, and L. Huang. 2009. A pig-human comparative RH map comprising 20 genes on pig chromosome 13q41 that harbours the ETEC F4ac receptor locus. J. Anim. Breed Genet. 126:30-36. https://doi.org/10.1111/j.1439-0388.2008.00751.x
  10. Sette, C., S. Dolci, R. Geremia, and P. RossiP. 2000. The role of stem cell factor and of alternative c-kit gene products in the establishment, maintenance and function of germ cells. Int. J. Dev. Biol. 44:599-608.
  11. Yan, N., Y. Lu, H. Sun, D. Tao, S. Zhang, W. Liu, and Y. Ma. 2007. A microarray for microRNA profiling in mouse testis tissues. Reproduction 134:73-79.. https://doi.org/10.1530/REP-07-0056
  12. Yoshida, S., Y. Nabeshima, and T. Nakagawa. 2007. Stem cell heterogeneity: actual and potential stem cell compartments in mouse spermatogenesis. Ann. N Y Acad. Sci. 1120:47-58. https://doi.org/10.1196/annals.1411.003
  13. Zhou, Y., Y. Liu, and L. Cheng. 2012. miR-21 expression is related to particle-induced osteolysis pathogenesis. J. Orthop. Res. 30:1837-1842. https://doi.org/10.1002/jor.22128