Analysis of gene expression in mouse 2-cell embryos using fluorescein differential display: comparison of culture environments.
The effect of the oviductal environment on gene expression in 2-cell mouse embryos was examined with mRNA differential display. Embryos used for experiments were cultured in modified Whitten medium with or without oviductal tissue until late 2-cell stage. The results of sequencing indicated that the genes for ATP synthase (ATPase 6), S:-adenosylmethionine decarboxylase (S:-AMDC) and nuclear autoantigenic sperm protein (NASP) were differentially expressed in embryos cultured in the oviductal environment (nonblocking culture condition). The ATPase 6 gene is encoded by mitochondrial DNA and is essential for the production of ATP. This indicates that the expression of ATP synthesis-related genes at the 2-cell stage may be required to maintain normal development in vitro. S:-Adenosylmethionine decarboxylase decarboxylates adenosylmethionine, which is a substrate of DNA methylation. The expression of S:-AMDC may be responsible for the low level of methylation of preimplantation development. As NASP is a histone-binding protein that is thought to be testis and sperm specific, its function in embryos remains unclear. On the other hand, the Tcl1 gene and a novel gene, the c-1 gene, were strongly expressed in embryos cultured without oviductal tissue (blocking culture condition). The expression patterns of these genes are quite similar. However, the detailed functions of these genes in embryos remain to be determined.
Minami
N
N
Laboratory of Reproductive Physiology, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan. naojiro@kais.kyoto-u.ac.jp
Sasaki
K
K
Aizawa
A
A
Miyamoto
M
M
Imai
H
H
D000251:Adenosine Triphosphatases / Q000235:genetics; D000255:Adenosine Triphosphate / Q000096:biosynthesis; D012437:Adenosylmethionine Decarboxylase / Q000235:genetics; D000818:Animals; D001324:Autoantigens / Q000235:genetics; D001755:Blastocyst / Q000378:metabolism; D018797:Cell Cycle Proteins; D046508:Culture Techniques; D019175:DNA Methylation; D004586:Electrophoresis; D004622:Embryo, Mammalian / Q000378:metabolism*; D005260:Female; D005307:Fertilization in Vitro; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D015870:Gene Expression; D051379:Mice; D008813:Mice, Inbred ICR; D009687:Nuclear Proteins / Q000235:genetics; D010448:Peptide Initiation Factors / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
D001324:Autoantigens; D018797:Cell Cycle Proteins; D005456:Fluorescent Dyes; D009687:Nuclear Proteins; D010448:Peptide Initiation Factors; C021019:eIF-6; C067535:Nasp protein, mouse; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; D012437:Adenosylmethionine Decarboxylase; D016650:Fluorescein-5-isothiocyanate