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Role of Messenger RNA Expression of Platelet Activating Factor and Its Receptor in Porcine In Vitro-Fertilized and Cloned Embryo Development¹

Department of Theriogenology and Biotechnololgy,³ College of Veterinary Medicine, Seoul National University, Seoul, 151-742, Korea Division of Biological Science,⁴ Gachon Medical School, Incheon, 417-840, Korea School of Agricultural Biotechnology,⁵ Seoul National University, Seoul, 151-742, Korea The Xenotransplantation Research Center,⁶ Seoul National University Hospital, Seoul, 110-744, Korea

Platelet activating factor (PAF) is known as an autocrine growth/survival factor in mammalian preimplantation embryos. This study investigated the expression of porcine PAF receptor (PAFr) mRNA and its role in porcine in vitro fertilized (IVF) or somatic cell nuclear transfer (SCNT) embryo development. The expression of PAFr mRNA in IVF or SCNT blastocysts was shown by reverse transcription-polymerase chain reaction (RT-PCR) and Southern blot analysis. Semiquantitative RT-PCR and Southern blot analysis demonstrated that PAFr mRNA was expressed during preimplantation embryo development, it was highly expressed through the 2-cell to 8-cell embryo stage, and it decreased at the morula stage. PAFr mRNA expression was detected steadily in IVF embryos, whereas it was varied at the 2-cell, 4-cell, and blastocyst stages in SCNT embryos. To determine the role of PAF in IVF and SCNT embryo development, embryos were cultured in North Carolina State University (NCSU)-23 medium supplemented with different concentrations of PAF (0, 0.037, 0.37, 3.72, or 37.2 nM). The PAF supplement significantly increased the rate of blastocyst formation in SCNT embryos, but not in IVF embryos. The PAF supplement for the entire 168 h of culture showed significantly higher blastocyst formation in SCNT embryos. Upregulation of PAFr mRNA by PAF in SCNT embryos indicated that the embryotrophic effect of PAF was mediated through its functional receptors in SCNT embryos. In conclusion, the present study demonstrated that PAFr mRNA was expressed in porcine IVF and SCNT embryos, and that PAF supplement improved the developmental competence of SCNT embryos through its specific receptors.

¹ Supported by grants M10310060000-03B4606-00000 from the Korean Ministry of Science and Technology and SC14031 from Stem Cell Research Center of the 21st Century Frontier Research Program funded by the Korean Ministry of Science and Technology. The authors acknowledge a graduate fellowship provided by the Ministry of Education through the BK21 program.

² Correspondence: Byeong Chun Lee, Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, Seoul 151-742, Korea. FAX: 82 2 884 1902; firstlee{at}snu.ac.kr

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