Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (23): 5037-5043.doi: 10.3864/j.issn.0578-1752.2013.23.021

• ANIMAL SCIENCE·VETERINARY SCIENCERE·SOURCE INSECT • Previous Articles     Next Articles

Ovarian Immunohistochemistry Localization and Preantral Follicles Expression Pattern Analysis of Buffalo Proliferating Cell Nuclear Antigen

 MA  Fan, ZHOU  Yu, GAO  Ya-Ke, ZHU  Peng, LEI  Xiao-Can, LIU  Qing-You, SHI  De-Shun   

  1. State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bio Resources, Guangxi University, Nanning 530005
  • Received:2013-04-19 Online:2013-12-01 Published:2013-07-26

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Abstract: 【Objective】To explore the involvement of nuclear antigen protein (PCNA) duiring different periods of buffalo follicular genesis, the expression pattern and ovary immunohistochemistry localization was performed in the present study.【Method】The localization of PCNA was demonstrated in ovarian tissue through the method of paraffin sections with immunohistochemistry, the mRNA expression level of PCNA in preantral follicles of buffalo was determined by real-time fluorescence quantification PCR(QRT-PCR).【Result】The immunohistochemical results showed that, in primordial follicle, no remarkable staining for PCNA in granulosa cells was observed, except the staining of nuclei in oocyte. In primary to secondary follicle, positive staining in oocytes and in some granulosa cells was observed, and PCNA immunoreactivity in granulosa cells of secondary follicle was obviously enhanced than that of primary follicles.The antral follicles showed extensive PCNA labeling in the layers of granulosa and theca cells and in the cumulus cells encircling the oocyte. The results of QRT-PCR showed that,the expression of PCNA in primordial follicles, primary follicles and secondary follicles showed a upregulation trend (43.50333±0.138338 vs 1.633804±0.093796 vs 1±0.012219 , P＜0.01).【Conclusion】In buffalo primary follicles, the expression of PCNA could be detected in granulosa cells, and was obviously enhanced following the development of follicle. The expression pattern of PCNA determined by QRT-PCR in preantral follicles was accordant with the results of immunohistochemistry. These results will provide a foundation for clarifing the mechanism of PCNA involving of follicular genesis in Buffalo.

Key words: Buffalo , PCNA , immunohistochemistry , quantitative PCR assay

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