Sensitivity of bovine blastocyst gene expression patterns to culture environments assessed by differential display RT-PCR.
The use of culture media to support the development of preimplantation embryos to the blastocyst stage is often associated with detrimental effects on normal development. These effects have been uncovered largely by investigating the phenotypic abnormalities displayed by fetuses and newborns derived from cultured preimplantation embryos. Research to understand the impact of culture on the embryonic developmental programme has focused on embryo metabolism, gene expression and genomic imprinting. We have used differential display RT-PCR to examine culture influences on global transcript pools in bovine embryos. Others have examined culture influences on candidate "marker genes" in cultured murine, ovine and bovine embryos. These studies have demonstrated that culture conditions influence the amount of marker gene transcripts and downregulate or induce the expression of novel genes during early development. Optimized defined culture media maintain embryonic gene expression patterns closely resembling those displayed by embryos derived in vivo. Preimplantation mammalian embryos display an impressive capacity to respond to the pressures that suboptimal culture environments place upon them. However, this plasticity operates within a defined range of tolerances. Continued research using molecular techniques will lead to increased understanding of developmental mechanisms causing culture-related phenotypic abnormalities in post-implantation embryos.
Natale
D R
DR
Department of Obstetrics and Gynaecology and Department of Physiology, The University of Western Ontario, London, Ontario, Canada.
De Sousa
P A
PA
Westhusin
M E
ME
Watson
A J
AJ
D000818:Animals; D001755:Blastocyst / Q000502:physiology*; D002417:Cattle; D002478:Cells, Cultured; D003001:Cloning, Molecular; D003470:Culture Media / Q000494:pharmacology*; D018076:DNA, Complementary / Q000032:analysis; D047108:Embryonic Development / Q000187:drug effects; D005314:Embryonic and Fetal Development / Q000235:genetics*; D005260:Female; D015870:Gene Expression; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects*; D005865:Gestational Age; D051379:Mice; D011247:Pregnancy; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015053:Zygote / Q000502:physiology
D003470:Culture Media; D018076:DNA, Complementary; D012333:RNA, Messenger