Basic FGF Promotes Proliferation of Ovarian Granulosa Cells in the Laying Chickens Via FGFR1 and PKC Pathway

Authors

  • J Lin,

    1. Key Laboratory of Animal Epidemic Etiology & Immunological Prevention of the Ministry of Agriculture, Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou, China
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  • Y Jia,

    1. Key Laboratory of Animal Epidemic Etiology & Immunological Prevention of the Ministry of Agriculture, Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou, China
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  • W Zeng,

    1. Key Laboratory of Animal Epidemic Etiology & Immunological Prevention of the Ministry of Agriculture, Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou, China
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  • Y Mi,

    1. Key Laboratory of Animal Epidemic Etiology & Immunological Prevention of the Ministry of Agriculture, Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou, China
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  • C Zhang

    1. Key Laboratory of Animal Epidemic Etiology & Immunological Prevention of the Ministry of Agriculture, Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou, China
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Author’s address (for correspondence): Caiqiao Zhang, College of Animal Sciences, Zhejiang University, No. 866 Yuhangtang Road, Hangzhou 310058, China. E-mail: cqzhang@zju.edu.cn

Contents

The proliferating effect of basic fibroblast growth factor (bFGF) on granulosa cells from ovarian pre-hierarchical follicles was evaluated in the laying chickens. Expression of bFGF receptor 1 (FGFR1) from small yellow follicles was determined by immunohistochemistry and RT-PCR. The FGFR1 protein and mRNA were intensively expressed in the granulosa layer. After 8- to 24-h treatment with bFGF (0.1–100 ng/ml), the proliferation of cultured granulosa cells was remarkably enhanced in a dose- and time-dependent manner. The FGFR1 antagonist SU5402 inhibited bFGF-induced cell proliferation. This stimulating effect was further confirmed by 5-bromo-2-deoxyuridine incorporation and terminal transferase dUTP nick end-labelling assay. Immunocytochemistry of protein kinases A (PKA) and C (PKC) showed that the pro-proliferation action of bFGF predominantly activated PKC expression. Meanwhile, the bFGF-induced cell proliferation was significantly promoted by PKC activator PMA and inhibited by PKC inhibitor H7 (p < 0.05). In addition, the bFGF-elicited cell proliferation was accompanied with increased mRNA expression of the cell cycle-regulating genes including cyclins D1 and E1, cyclin-dependent kinases 2 and 6. In conclusion, bFGF promoted the proliferation of ovarian granulosa cells through binding with FGFR1 and involving PKC pathway in the pre-hierarchical follicles of the laying chickens.

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