Developmental competence and gene expression in preimplantation bovine embryos derived from somatic cell nuclear transfer using different donor cells.
This study compared the developmental competence of somatic cell nuclear transfer (SCNT) embryos reconstructed with different donor cells and analysed gene expression in the resulting embryos. Bovine fetal/adult ear fibroblasts and cumulus cells were used as donor cells and the developmental competence of the reconstructed embryos was monitored. The cell number and allocation in blastocysts were determined by differential staining. The Bax, E-cad, IF-tau, Hsp (heat shock protein) 70, Igf2r (insulin-like growth factor 2 receptor), DNMT (DNA methyltransferase) 1 and Mash (mammalian achaete-scute homologue) 2 genes were selected for gene expression analysis. The relative abundance (ratio to GAPDH mRNA) of gene transcripts in blastocysts was measured by semiquantitative reverse transcription-polymerase chain reaction. In experiment 1, development of SCNT preimplantation embryos and the cell numbers of inner cell masses and trophoblasts were not different among SCNT embryos derived from different cell types. In experiment 2, the relative expression of GAPDH and Hsp 70 transcripts was similar in all embryos. The expression of Bax, Igf2r and Mash2 transcripts was significantly increased in SCNT embryos reconstructed with adult fibroblasts. The E-cad transcript levels were reduced in SCNT embryos reconstructed with fetal fibroblasts. Relative abundance of DNMT1 in SCNT embryos derived from fetal fibroblasts was increased, and IF-tau expression in SCNT embryos derived from cumulus cells was increased. In conclusion, depending on the type of donor cells, preimplantation SCNT embryos displayed marked differences in gene expression. This may affect the developmental competence of SCNT embryos reconstructed with different cell types after implantation or during fetal growth in vivo.
Jang
Goo
G
Department of Theriogenology and Biotechnology, College of Veterinary Medicine and School of Agricultural Biotechnology, Seoul National University, Korea.
Jeon
Hyun Yong
HY
Ko
Kyung Hee
KH
Park
Hee Jung
HJ
Kang
Sung Keun
SK
Lee
Byeong Chun
BC
Hwang
Woo Suk
WS
D000818:Animals; D001755:Blastocyst / Q000378:metabolism*; D002417:Cattle / Q000235:genetics*; D002478:Cells, Cultured; D019976:Cloning, Organism / Q000379:methods*; D047108:Embryonic Development / Q000235:genetics*; D005260:Female; D015870:Gene Expression; D018507:Gene Expression Regulation, Developmental; D012333:RNA, Messenger / Q000378:metabolism; D014158:Transcription, Genetic
D012333:RNA, Messenger