Pregnancy-associated changes in genome-wide gene expression profiles in the liver of cow throughout pregnancy.
The objective of the present study was to fabricate and use a bovine liver complementary DNA (cDNA) microarray to profile genome-wide gene expressions in the liver of cow throughout pregnancy. A cDNA library was prepared from liver total RNA collected from cows during estrous cycle and pregnancy, and from fetuses at different stages of pregnancy. The sequenced clones were compiled and annotated by basic local alignment search tool (BLASTn) and spotted onto glass slides. The annotated liver array represented 2675 genes. Of which, 1442 were known genes while 617 sequences had matches with sequences found in expressed sequence tags databases. In addition, 616 unknown sequences were found and these sequences may possibly be identified as candidates for novel bovine genes. For gene expression profiling studies, total RNA from livers of cows slaughtered on days 19, 27-28, 49-58, 150, and 245 of pregnancy (test RNAs) was separately reverse transcribed and labeled with either cyanine 5-fluorescent dye (Cy5) or Cy3. The test samples were individually compared with liver total RNA collected from nonpregnant cycling cows (control RNA) after reverse transcription and labeling with the opposite dye following a two-color hybridization method. After scanning, image acquisition, and normalization, genes that showed either more than 1.5-fold (test/control) induction or repression were selected for further analyses. Hierarchical clustering algorithm showed a clear induction of most liver genes on days 27-28 of pregnancy. Self-organizing maps algorithm identified groups of genes whose differential expression patterns were similar across pregnancy. In conclusion, we described fabrication of a bovine liver cDNA microarray, and demonstrate, for the first time, differential expression patterns of a large number of coregulated liver genes in parallel throughout pregnancy in the bovine.
Herath
Chandana B
CB
Laboratory of Reproductive Biology and Technology, National Institute of Agrobiological Sciences, Tsukuba City, Ibaraki 305-8602, Japan.
Shiojima
Satoshi
S
Ishiwata
Hiroko
H
Katsuma
Susumu
S
Kadowaki
Tadashi
T
Ushizawa
Koichi
K
Imai
Kei
K
Takahashi
Toru
T
Hirasawa
Akira
A
Tsujimoto
Gozoh
G
Hashizume
Kazuyoshi
K
D000465:Algorithms; D000818:Animals; D002417:Cattle; D018076:DNA, Complementary / Q000378:metabolism; D019992:Databases as Topic; D020224:Expressed Sequence Tags; D005260:Female; D016678:Genome; D008099:Liver / Q000378:metabolism; D005810:Multigene Family; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D011247:Pregnancy; D011270:Pregnancy, Animal; D012313:RNA / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012984:Software; D013997:Time Factors
D018076:DNA, Complementary; D012313:RNA