Comparison of gene expression in individual preimplantation bovine embryos produced by in vitro fertilisation or somatic cell nuclear transfer
Luiz Sergio de A. Camargo A B , Anne M. Powell B , Vicente R. do Vale Filho C and Robert J. Wall C D
+ Author Affiliations
- Author Affiliations
A Embrapa Dairy Cattle, Rua Eugênio do Nascimento, 610, Juiz de Fora, MG, 36038-330 Brazil.
B School of Veterinary Science, Federal University of Minas Gerais, Av. Antôno Carlos, 6627, Belo Horizonte, MG, 30123-970 Brazil.
C Biotechnology and Germplasm Laboratory, Agricultural Research Service, Beltsville Agricultural Research Center-EAST, Building 200, Beltsville, MD 20705-2350, USA.
D Corresponding author. Email: bobwall@anri.barc.usda.gov
Reproduction, Fertility and Development 17(5) 487-496 https://doi.org/10.1071/RD04128
Submitted: 2 November 2004 Accepted: 13 March 2005 Published: 2 May 2005
Abstract
In vitro fertilisation (IVF) and somatic cell nuclear transfer (SCNT) have been implicated in a variety of developmental abnormalities. Aberrant gene expression is likely to account for much of the diminished viability and developmental abnormalities observed. In the present study, the expression of multiple genes in IVF and SCNT bovine blastocyst-stage embryos were evaluated and compared with in vivo-produced embryos. Eleven genes expressed at and following maternal–zygotic transcription transition were evaluated in individual blastocysts by real-time polymerase chain reaction following RNA amplification. A subset of those genes was also evaluated in individual IVF and SCNT eight-cell embryos. A fibroblast-specific gene, expressed by nuclear donor cells, was also evaluated in IVF and SCNT embryos. The observed gene expression pattern at the eight-cell stage was not different between IVF and SCNT embryos (P > 0.05). In vitro fertilisation and SCNT blastocyst expression was lower (P < 0.01) for all genes compared with their in vivo-produced counterparts, except for lactate dehydrogenase isoenzyme A (P < 0.001). The patterns of gene expression of the IVF and SCNT blastocysts were indistinguishable. Neither SCNT eight-cell nor blastocyst-stage embryos expressed the gene used as a fibroblast marker (collagen VIα1). For the genes evaluated, the level of expression was influenced more by the environment than by the method used to produce the embryos. These results support the notion that if developmental differences observed in IVF- and SCNT-produced fetuses and neonates are the result of aberrant gene expression during the preimplantation stage, those differences in expression are subtle.
Extra keywords: cloning, early development.
Acknowledgments
The authors thank Harold Hawk for performing the in vivo embryo collection and Paul Graninger for his assistance in the laboratory. This study was supported by the CAPES Foundation, Embrapa and ARS/USDA.
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