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Absence of Donor-Derived Zona Pellucida Protein C Homolog in the Inner Perivitelline Layer of Peking Duck (Anas platyrhynchos)-Japanese Quail (Coturnix coturnix japonica) Chimeras (Duails)¹

J. Sha^*, J. Gao^*, J. Li^*, Q. Zhao^*, G. Tao^*, C. Zhao^*, H. Han^*, M. Mori and Z. Li^*,2

^* Department of Biochemistry and Molecular Biology, College of Biological Science, China Agricultural University, Beijing 100094, China; and Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, Shizuoka 422-8529, Japan

² Corresponding author: lzdws{at}cau.edu.cn

Avian blastodermal cells at stage X are used to produce interspecies chimeras for heterogenous poultry reproduction. However, recipient-derived inner perivitelline layer (IPVL)-enclosed donor-derived ova may affect the efficiency of germline transmission via chimera. Among the proteins in the IPVL, zona pellucida protein C (ZPC) plays an important role in sperm-egg binding and inducing the acrosome reaction. In the present study, Peking duck blastodermal cells at stage X were transferred into subgerminal cavities of Japanese quail embryos at the same stage. Fourteen female duck-quail chimeras (duails) were hatched and raised to sexual maturity. After being screened by PCR, 3 duails were selected for examination of donor-derived ZPC. A total of 152 IPVL protein samples from the individual eggs laid by the 3 duails then underwent a preliminary examination for the presence of donor-derived ZPC by means of SDS-PAGE, periodic acid-Schiff staining, and Western blotting. A novel 35-kDa ZPC, not observed in quail but in duck, was found in the IPVL of the duails. Further analysis of peptide mass fingerprinting of Peking duck ZPC, Japanese quail ZPC, and the 35-kDa duail ZPC by matrix-assisted laser desorption-ionization reflectron time-of-flight mass spectrometry revealed that the novel ZPC was an isoform of quail ZPC. Moreover, comparison of N-terminal amino acid sequences of these 3 ZPC confirmed that the 35-kDa quail ZPC had more amino acids at the N terminus than did native quail ZPC, and none of the donor-derived ZPC was found in the duails. These findings suggest that it would be difficult to obtain donor-derived offspring by natural mating of interspecies chimeras.

Key Words: zona pellucida protein C • blastodermal cell • Peking duck • chimera • matrix-assisted laser desorption-ionization reflectron time-of-flight mass spectrometry

¹ Supported by grant-in-aids for scientific research (30370721) from the National Natural Science Foundation of China, Beijing.