Microarray analysis predicts differentially expressed genes in theca cells from cows with high intrafollicular androstenedione are regulated by ESR1 and VEGFA signaling and increased mRNA stability

Authors: BOCHANTIN, KERRI - University Of Nebraska; SUMMERS, ADAM - New Mexico State University; POHLMEIER, WILLIAM - University Of Nebraska; SARGENT, KEVIN - University Of Nebraska; KURZ, SCOTT - University Of Nebraska; ROMEREIM, SARAH - University Of Nebraska; DAUDU, OLUREMI - University Of Nebraska; MCFEE, RENEE - University Of Nebraska; Cushman, Robert - Bob; DAVIS, JOHN - Nebraska Medical Center; CUPP, ANDREA - University Of Nebraska; WOOD, JENNIFER - University Of Nebraska

Submitted to: Society for the Study of Reproduction Annual Meeting
Publication Type: Abstract Only
Publication Acceptance Date: 5/1/2018
Publication Date: 7/2/2018
Citation: Bochantin, K.A., Summers, A.F., Pohlmeier, W.E., Sargent, K.M., Kurz, S.G., Romereim, S.M., Daudu, O., Mcfee, R.M., Cushman, R.A., Davis, J.S., Cupp, A.S., Wood, J.R. 2018. Microarray analysis predicts differentially expressed genes in theca cells from cows with high intrafollicular androstenedione are regulated by ESR1 and VEGFA signaling and increased mRNA stability [abstract]. Society for the Study of Reproduction Annual Meeting. Abstract Program p. 327-328. Available: https://www.ssr.org/sites/ssr.org/files/2018_annual_meeting_abstracts_updated.pdf

Interpretive Summary:

Technical Abstract: We identified a population of cows in the UNL physiology herd with high androstenedione (High A4; >40 ng/ml) concentrations in the follicular fluid of the dominant follicle. These High A4 cows display irregular estrous cycles and tended to have reduced calving rates indicating that this phenotype impairs fertility. The molecular phenotype of theca cells from High A4 cows includes increased expression of steroidogenic enzymes (CYP11A1, CYP17A1) and LHCGR. In the current study we hypothesized that theca cells from High A4 cows have a unique gene expression profile, which contributes to the altered function including increased androgen synthesis by these cells. To test this hypothesis, estrous cycles of High A4 and Control cows were synchronized with an injection of GnRH and a controlled internal drug release device (CIDR). After 7 days, the CIDR was removed, an injection of prostaglandin F2 alpha was given, and ovariectomies performed 36 hours later. Theca cells were microdissected from the dominant estrogen-active follicle and RNA extracted. High quality RNA from High A4 (n = 5) and control (n = 4) cows were labeled and hybridized to Affymetrix Bovine GeneChip Gene 1.0 ST Arrays at the University of Nebraska Microarray Core facility. Following hybridization, normalized data was generated using Robust Multi-Array Averaging Whole transcriptome analysis. Normalized data were subsequently analyzed using the NIA Array Analysis tool. Hierarchical clustering showed that samples from individual cows clustered based on A4 classification. Analysis of Variance (ANOVA, P 2) included the mitotic roles of Polo-Like Kinases, the role of BRCA1 in DNA damage response, and Rho Family GTPase signaling. Conversely, p53 signaling was inhibited (Z-score Interestingly, several predicted regulators play an important role in mRNA stability. Specifically, ELAVL1, an RNA binding protein that stabilizes mRNAs, was activated whereas ZFP36, an RNA binding protein that destabilizes mRNAs was inhibited. In addition, microRNA families with predicted inhibition included mir-21, let-7, miR 291a-3p, and miR-483-3p. The alter expression of these important signaling and stability factors may contribute to the altered steroidogenesis and delayed follicular development phenotype expressed by High A4 cows.