Donor Cell Source (Miniature Pig and Landrace Pig) Affects Apoptosis and Imprinting Gene Expression in Porcine Nuclear Transfer Embryos
2008
Park, M.R. (National Institute of Animal Science, RDA, Suwon, Republic of Korea) | Hwang, I.S. (National Institute of Animal Science, RDA, Suwon, Republic of Korea) | Shim, J.H. (National Institute of Animal Science, RDA, Suwon, Republic of Korea) | Moon, H.J. (National Institute of Animal Science, RDA, Suwon, Republic of Korea) | Kim, D.H. (National Institute of Animal Science, RDA, Suwon, Republic of Korea) | Ko, Y.G. (National Institute of Animal Science, RDA, Suwon, Republic of Korea) | Seong, H.H. (National Institute of Animal Science, RDA, Suwon, Republic of Korea) | Im, G.S. (National Institute of Animal Science, RDA, Suwon, Republic of Korea), E-mail: [email protected]
This study investigated the developmental ability and gene expression of somatic cell nuclear transfer embryos using ear skin fibroblast cells derived from miniature pig. When miniature pig (m) and landrace pig (p) were used as donor cells, there were no differences in cleavage (79.2 vs. 78.2%) and blastocyst rates (27.4 vs. 29.7%). However, mNT blastocysts showed significantly higher apoptosis rate than that of pNT blastocysts (6.1 vs. 1.7%) (p less than 0.05). The number of nuclei in pNT blastosysts was significantly higher than that of mNT (35.8 vs. 29.3) (p less than 0.05). Blastocysts were analyzed using Realtime RT-PCR to determine the expression of Bax-α, Bcl-xl, H19, IGF2, IGF2r and Xist. Bax-α was higher in mNT blastocyst than pNT blastocyst (p less than 0.05). There was no difference in Bcl-xl between two NT groups. Bax-α/Bcl-xl was, however, significantly higher in mNT blastocyst compared to pNT. The expression of imprinting genes were aberrant in blastocysts derived from NT compared to in vivo blastocysts. H19 and IGF2r were significantly lower in mNT blastocysts (p less than 0.05). The expression of IGF2 and Xist was similar in two NT groups. However, imprinting genes were expressed aberrantly in mNT compared to pNT blastocysts. The present results suggest that the NT between donor cells derived from miniature pig and recipient oocytes derived from crossbred pig might affect reprogramming of donor cell, resulting in high apoptosis and aberrant expression patterns of imprinting genes.
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