Deep sequencing of the transcriptome in the anterior pituitary of heifers before and after ovulation

Abstract

We aimed to determine gene expression patterns in the anterior pituitary (AP) of heifers before and after ovulation via deep sequencing of the transcriptome (RNA-seq) to identify new 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. We randomly selected 4 pre-ovulation and 4 post-ovulation APs, and the ribosomal RNA-depleted poly (A)+RNA were prepared to assemble next-generation sequencing libraries. The bovine APs expressed 12,769 annotated genes at pre- or post-ovulation. The sum of the reads per kilobase of exon model per million mapped reads (RPKM) values of all transcriptomes were 599,676 ± 38,913 and 668,209 ± 23,690, and 32.2 ± 2.6% and 44.0 ± 4.4% of these corresponded 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 included two G-protein-coupled receptor (GPCR) genes (GPR61 and GPR153) and those encoding 13 binding proteins. The AP also expressed 259 receptor and other 364 binding proteins. Moreover, ingenuity pathway analysis for the 396 genes revealed (P=2.4 × 10−3) a canonical pathway linking GPCR to cytoskeleton reorganization, actin polymerization, microtubule growth, and gene expression. Thus, the present study clarified the novel genes found to be differentially expressed before and after ovulation and clarified an important pathway in the AP.

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“…Furthermore, in our study, we found lncRNA MSTRG.9440.3 target GPR61 , SORT1 , SYPL2 and PSMA5 in sheep pituitary gland. Previous studies suggested that GPR61 has a different expression in the pre- and post-ovulation of bovine anterior pituitary [ 62 ]. Although the ligand(s) and functions of GPR61 are not clear, it is known to associate with the Gs protein and stimulate extracellular regulated protein kinases ( ERK ) signaling in neurons [ 63 , 64 ].…”

A novel somatic mutation of SIN3A detected in breast cancer by whole-exome sequencing enhances cell proliferation through ERα expression

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Discovery of new receptors regulating luteinizing hormone and follicle-stimulating hormone secretion by bovine gonadotrophs to explore a new paradigm for mechanisms regulating reproduction

“…My previous deep transcriptome sequencing studies in heifer AP before and after ovulation did not reveal significant differences in the expression of AMHR2, the receptor for AMH [40]. AMH is a dimeric glycoprotein belonging to the transforming growth factor (TGF)-β family and it is primarily produced by the granulosa cells of the preantral and small antral follicles in humans and animals [51].…”

“…Therefore, I assessed gene expression patterns via RNA-seq in the AP, but not in the purified gonadotrophs of heifers before and after ovulation [40]. Consequently, 396 differentially expressed genes were identified in the bovine AP (P < 0.05) in pre-and post-ovulation samples, which included two GPCR genes (GPR61 and GPR153), and nine receptors for previously identified ligands.…”