Effect of in vitro fertilization on gene expression and development of mouse preimplantation embryos.
In vitro culture (IVC) of preimplantation mouse embryos is associated with changes in gene expression. It is however, not known if the method of fertilization affects the global pattern of gene expression. We compared gene expression and development of mouse blastocysts produced by in vitro fertilization (IVF) versus blastocysts fertilized in vivo and cultured in vitro from the zygote stage (IVC) versus control blastocysts flushed out of the uterus on post coital day 3.5. The global pattern of gene expression was assessed using the Affymetrix 430 2.0 chip. It appears that each method of fertilization has a unique pattern of gene expression and development. Embryos cultured in vitro had a reduction in the number of trophoblastic cells (IVF 33.5 cells, IVC 39.9 cells, and 49.6 cells in the in vivo group) and, to a lesser degree, of inner cell mass cells (12.8, 11.7, and 13.8 respectively). The inner cell mass nuclei were larger after culture in vitro (140 microm(2), 113 microm(2), and 86 microm(2) respectively). Although a high number of genes (1912) was statistically different in the IVF cohort when compared with the in vivo control embryos, the magnitude of the changes in gene expression were low and only a minority of genes (29 genes) was changed more than fourfold. Surprisingly, IVF embryos were different from IVC embryos (3058 genes were statistically different, but only three changed more than fourfold). Proliferation, apoptosis, and morphogenetic pathways are the most common pathways altered after IVC. Overall, IVF and embryo culture have a profound effect on gene expression pattern and phenotype of mouse preimplantation embryos.
Giritharan
Gnanaratnam
G
Department of Obstetric and Gynecology, University of California, San Francisco, San Francisco, California 94143, USA.
Talbi
Said
S
Donjacour
Annemarie
A
Di Sebastiano
Francesca
F
Dobson
Anthony T
AT
Rinaudo
Paolo F
PF
D000818:Animals; D017209:Apoptosis; D001755:Blastocyst / Q000166:cytology / Q000502:physiology*; D002452:Cell Count; D049109:Cell Proliferation; D002478:Cells, Cultured; D047108:Embryonic Development; D005260:Female; D005306:Fertilization / Q000502:physiology; D005307:Fertilization in Vitro; D015870:Gene Expression; D020869:Gene Expression Profiling; D008297:Male; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis; D011247:Pregnancy; D013048:Specimen Handling; D015053:Zygote / Q000166:cytology / Q000502:physiology