Deep sequencing of the transcriptome in the anterior pituitary of heifersbefore and after ovulation

[摘要] We aimed to determine gene expression patterns in the anterior pituitary (AP) of heifersbefore and after ovulation via deep sequencing of the transcriptome (RNA-seq) to identifynew genes and clarify important pathways. Heifers were slaughtered on the estrus day(pre-ovulation; n=5) or 3 days after ovulation (post-ovulation; n=5) for AP collection. Werandomly selected 4 pre-ovulation and 4 post-ovulation APs, and the ribosomal RNA-depletedpoly (A)+RNA were prepared to assemble next-generation sequencing libraries. The bovineAPs expressed 12,769 annotated genes at pre- or post-ovulation. The sum of the reads perkilobase of exon model per million mapped reads (RPKM) values of all transcriptomes were599,676 ± 38,913 and 668,209 ± 23,690, and 32.2 ± 2.6% and 44.0 ± 4.4% of thesecorresponded to the AP hormones in the APs of pre- and post-ovulation heifers,respectively. The bovine AP showed differential expression of 396 genes(P<0.05) in the pre- and post-ovulation APs. The 396 genes includedtwo G-protein-coupled receptor (GPCR) genes (GPR61 andGPR153) and those encoding 13 binding proteins. The AP also expressed259 receptor and other 364 binding proteins. Moreover, ingenuity pathway analysis for the396 genes revealed (P=2.4 × 10−3) a canonical pathway linkingGPCR to cytoskeleton reorganization, actin polymerization, microtubule growth, and geneexpression. Thus, the present study clarified the novel genes found to be differentiallyexpressed before and after ovulation and clarified an important pathway in the AP.

[学科分类] 兽医学

[关键词] G-protein-coupled receptor;Rho family GTPase;RNA-seq;ruminant