title: Pregnancy-associated changes in genome-wide gene expression profiles in the liver of cow throughout pregnancy.
abstract: 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.
authors.affiliation: Laboratory of Reproductive Biology and Technology, National Institute of Agrobiological Sciences, Tsukuba City, Ibaraki 305-8602, Japan.
authors.forename: Chandana B
authors.initials: CB
authors.lastname: Herath
authors.forename: Satoshi
authors.initials: S
authors.lastname: Shiojima
authors.forename: Hiroko
authors.initials: H
authors.lastname: Ishiwata
authors.forename: Susumu
authors.initials: S
authors.lastname: Katsuma
authors.forename: Tadashi
authors.initials: T
authors.lastname: Kadowaki
authors.forename: Koichi
authors.initials: K
authors.lastname: Ushizawa
authors.forename: Kei
authors.initials: K
authors.lastname: Imai
authors.forename: Toru
authors.initials: T
authors.lastname: Takahashi
authors.forename: Akira
authors.initials: A
authors.lastname: Hirasawa
authors.forename: Gozoh
authors.initials: G
authors.lastname: Tsujimoto
authors.forename: Kazuyoshi
authors.initials: K
authors.lastname: Hashizume
mesh_terms: 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
chemical_list: D018076:DNA, Complementary; D012313:RNA