In vitro and in vivo culture effects on mRNA expression of genes involved in metabolism and apoptosis in bovine embryos
Hiemke M. Knijn A C , Christine Wrenzycki B , Peter J. M. Hendriksen A , Peter L. A. M. Vos A , Elly C. Zeinstra A , Gijsbert C. van der Weijden A , Heiner Niemann B and Steph J. Dieleman AA Department of Farm Animal Health, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 7, 3584 CL Utrecht, the Netherlands.
B Department of Biotechnology, Institute for Animal Breeding (FAL), Neustadt, Germany.
C Corresponding author. Email: h.knijn@vet.uu.nl
Reproduction, Fertility and Development 17(8) 775-784 https://doi.org/10.1071/RD05038
Submitted: 30 March 2005 Accepted: 25 September 2005 Published: 16 December 2005
Abstract
Bovine blastocysts produced in vitro differ substantially from their in vivo-derived counterparts with regard to glucose metabolism, level of apoptosis and mRNA expression patterns. Maternal embryonic genomic transition is a critical period in which these changes could be induced. The goals of the present study were twofold: (1) to identify the critical period of culture during which the differences in expression of gene transcripts involved in glucose metabolism are induced; and (2) to identify gene transcripts involved in apoptosis that are differentially expressed in in vitro- and in vivo-produced blastocysts. Relative abundances of transcripts for the glucose transporters Glut-1, Glut-3, Glut-4 and Glut-8, and transcripts involved in the apoptotic cascade, including BAX, BCL-XL, XIAP and HSP 70.1, were analysed by a semiquantitative reverse transcription–polymerase chain reaction assay in single blastocysts produced in vitro or in vivo for specific time intervals, that is, before or after maternal embryonic transition. Whether the culture environment was in vitro or in vivo affected the expression of glucose transporter transcripts Glut-3, Glut-4 and Glut-8. However, the critical period during culture responsible for these changes, before or after maternal embryonic transition, could not be determined. With the exception of XIAP, no effects of culture system on the mRNA expression patterns of BAX, BCL-XL and HSP 70.1 could be observed. These data show that expression of XIAP transcripts in expanded blastocysts is affected by in vitro culture. These findings add to the list of bovine genes aberrantly expressed in culture conditions, but do not support the hypothesis that maternal embryonic transition is critical in inducing the aberrations in gene expression patterns studied here.
Acknowledgments
We are grateful to Christine Oei, Thea Blankenstein, Henk Heuveling, Klaus-Gerd Hadeler and Doris Herrmann for excellent technical assistance, and to the animal handlers for management of the animals. The critical reading of the manuscript by Ms Jane Collins, Boston, is highly appreciated. The authors thank Holland Genetics (Arnhem, the Netherlands) for supplying SOF culture medium. The Netherlands Organization for Scientific Research supported this study.
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