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http://dx.doi.org/10.5713/ajas.2010.90249

Comparison of Gene Expression between Cumulus Oocyte Complexes and Naked Oocytes by Suppression Subtractive Hybridization in Swine  

Xiang, Zhi Feng (Department of Animal Science, Henan Institute of Science and Technology)
Zhang, Jin Zhou (Department of Animal Science, Henan Institute of Science and Technology)
Li, Xue Bin (Department of Animal Science, Henan Institute of Science and Technology)
Xie, Hong Bin (Department of Animal Science, Henan Institute of Science and Technology)
Wang, Qing Hua (Department of Animal Science, Henan Institute of Science and Technology)
Publication Information
Asian-Australasian Journal of Animal Sciences / v.23, no.1, 2010 , pp. 17-24 More about this Journal
Abstract
In the antral follicle phase, several layers of cumulus cells surround the oocyte and play an important support and regulation role in oocyte development and maturation via intercellular communications and interactions between oocytes and cumulus cells. However, information on stage specific gene expression in swine during the phase is not well understood. To investigate the function of cumulus cells during in vitro maturation of porcine oocytes and gene expression, suppression subtractive hybridization (SSH) was performed to screen genes that were differentially expressed between cumulus-oocyte complexes (COCs) and naked oocytes (NOs). Utilizing mRNAs from in vitro maturation oocytes, a SSH cDNA library from COCs as the tester and NOs as the driver was constructed. The SSH cDNA library was then screened using dot blot analysis. Results showed that a total of 70 clones randomly selected from the library were differentially expressed. Among these, 41 exhibited high homology to known genes and 11 were novel expressed sequences tags (ESTs). Four differentially expressed genes, including bfgf, sprouty 2, egr and btc, were further studied by real time quantitative PCR; results confirmed an increased expression of respective mRNA in COCs compared with NOs, which suggests that these factors may play an important role in oocyte development and maturation.
Keywords
Suppression Subtractive Hybridization (SSH); Gene Expression; Cumulus Cells, Porcine;
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