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Early temporal transcriptome responses of the bovine mid-cycle corpus luteum to prostaglandin F2 alpha indicates cytokine activation

ABSTRACT: In ruminants, prostaglandin F2alpha (PGF2α)-mediated luteolysis is essential for resumption of the estrous cycle and is a target for improving fertility. To deduce the early PGF2α-provoked signaling elicited in the corpus luteum a time-course from 0.5–4 h was performed on cows at mid-cycle. A microarray-determined transcriptome was established and analyzed by Ingenuity Pathway Analysis (IPA). Self-organizing map analysis grouped differentially expressed transcripts into ten mRNA expression patterns indicative of signaling cascades. Classic PGF2α signaling (cyclic adenosine monophosphate, mitogen-activated protein kinases) and cytokine signaling (nuclear factor κB, interleukin) were both predicted by IPA after PGF2α administration. Comparison with two analogous datasets revealed a conserved group of 124 transcripts similarly changed in each dataset, which also predicted cytokine signaling. Elevated levels of cytokine transcripts after PGF2α and IPA-predicted activation of cytokine pathways demonstrate the importance of inflammatory reactions early in PGF2α -mediated luteolysis.

ORGANISM(S): Bos taurus

PROVIDER: GSE94069 | GEO | 2017/01/26

SECONDARY ACCESSION(S): PRJNA368717

REPOSITORIES: GEO

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Project description:Luteolysis of the corpus luteum (CL) during non-fertile cycles involves a cessation of progesterone (P4) synthesis (functional regression) and subsequent structural remodeling. The molecular processes responsible for initiation of luteal regression in the primate CL are poorly defined. Therefore, a genomic approach was utilized to systematically identify differentially expressed genes in the rhesus macaque CL during spontaneous luteolysis. CL were collected prior to (days 10-11 post-LH surge, mid-late [ML] stage) or during (days 14-16, late stage) functional regression. Based on P4 levels, late stage CL were subdivided into functional late (FL, serum P4 > 1.5 ng/ml) and functionally-regressed late (FRL, serum P4 < 0.5 ng/ml) groups (n=4 CL/group). Total RNA was isolated, labeled and hybridized to Affymetrix genome microarrays that contain elements representing the entire rhesus macaque transcriptome. With the ML stage serving as the baseline, there were 681 differentially expressed transcripts (>2-fold change; p< 0.05) that could be categorized into three primary patterns of expression: 1) increasing from ML through FRL, 2) decreasing from ML through FRL, and 3) increasing ML to FL, followed by a decrease in FRL. Ontology analysis revealed potential mechanisms and pathways associated with functional and/or structural regression of the macaque CL. Quantitative real-time PCR was used to validate microarray expression patterns of 13 genes with the results being consistent between the two methodologies. Protein levels were found to parallel mRNA profiles in 4 of 5 differentially expressed genes analyzed by Western blot. Thus, this database will facilitate the identification of mechanisms involved in primate luteal regression. Keywords: time course plus functional state of corpus luteum The first experimental factor is the stage of the luteal phase when the CL was collected. There were two stages: 1) between days 10-12 post LH-surge (mid-late stage), and 2) days 14-16 (late stage). The mid-late stage is a transitionary period where CL are still functional (i.e., producing significant quantities of P4) but are nearing the time when luteolysis initiates in non-conception cycles, and the late stage corresponds to the normal period when functional regression (cessation of P4 secretion) occurs in non-conception cycles. The second experimental factor is the functional state of the CL which was determined by measuring serum concentrations of P4 at the time of CL collection. Late stage CL with serum P4 levels > 1.5 ng/ml were considered to be functional, and those with serum P4 < 0.5 ng/ml were considered to have completed functional regression. Measuring mRNA levels in CL collected from these groups provides a comprehensive analysis of the primate transcriptome throughout the normal period of functional regression in the macaque CL. There were 12 biological replicates: n = 4 per group, 3 groups in total. The mid-late CL were used as the baseline reference for gene expression.

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Early temporal transcriptome responses of the bovine mid-cycle corpus luteum to prostaglandin F2 alpha indicates cytokine activation

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