Abnormal levels of transcript abundance of developmentally important genes in various stages of preimplantation bovine somatic cell nuclear transfer embryos.
Based on microarray data comparing gene expression of fibroblast donor cells and bovine somatic cell nuclear transfer (SCNT) and in vivo produced (AI) blastocysts, a group of genes including several transcription factors was selected for evaluation of transcript abundance. Using SYBR green-based real-time polymerase chain reaction (Q-PCR) the levels of POU domain class 5 transcription factor (Oct4), snail homolog 2 (Snai2), annexin A1 (Anxa1), thrombospondin (Thbs), tumor-associated calcium signal transducer 1 (Tacstd1), and transcription factor AP2 gamma (Tfap2c) were evaluated in bovine fibroblasts, oocytes, embryos 30 min postfusion (SCNT), 12 h postfertilization/activation, as well as two-cell, four-cell, eight-cell, morula, and blastocyst-stage in vitro fertilized (IVF) and SCNT embryos. For every gene except Oct4, levels of transcript were indistinguishable between IVF and SCNT embryos at the blastocyst stage; however, in many cases levels of these genes during stages prior to blastocyst differed significantly. Altered levels of gene transcripts early in development likely have developmental consequences downstream. These results indicate that experiments evaluating gene expression differences between control and SCNT blastocysts may underestimate the degree of difference between clones and controls, and further offer insights into the dynamics of transcript regulation following SCNT.
Department of Animal, Dairy, and Veterinary Sciences and Center for Integrated Biosystems, Utah State University, Logan, Utah 84322-4815, USA.
Kenneth I
KI
Aston
Guan-Peng
GP
Li
Brady A
BA
Hicks
Benjamin R
BR
Sessions
Aaron P
AP
Davis
Lee F
LF
Rickords
John R
JR
Stevens
Kenneth L
KL
White
D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000378:metabolism*; D002417:Cattle / Q000196:embryology / Q000235:genetics / Q000378:metabolism; D002467:Cell Nucleus / Q000235:genetics; D002478:Cells, Cultured; D046149:Embryo Culture Techniques / Q000662:veterinary; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism; D047108:Embryonic Development / Q000235:genetics*; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D015870:Gene Expression; D005796:Genes; D009028:Morula / Q000378:metabolism; D053652:Nuclear Transfer Techniques / Q000662:veterinary*; D009865:Oocytes / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics
D014157:Transcription Factors