A transcriptomal analysis of bovine oviductal epithelial cells collected during the follicular phase versus the luteal phase of the estrous cycle.

K L Cerny,P J Bridges,A.J Walton,L.H Anderson,E Garrett

doi:10.1186/s12958-015-0077-1

Abstract

BackgroundReproductive success depends on a functional oviduct for gamete storage, maturation, fertilization, and early embryonic development. The ovarian-derived steroids estrogen and progesterone are key regulators of oviductal function. The objective of this study was to investigate luteal and follicular phase-specific oviductal epithelial cell function by using microarray-based transcriptional profiling, to increase our understanding of mRNAs regulating epithelial cell processes, and to identify novel genes and biochemical pathways that may be found to affect fertility in the future.MethodsSix normally cycling Angus heifers were assigned to either luteal phase (LP, n = 3) or follicular phase (FP, n = 3) treatment groups. Heifers in the LP group were killed between day 11 and 12 after estrus. Heifers in the FP group were treated with 25 mg PGF2α (Lutalyse, Pfizer, NY) at 8 pm on day 6 after estrus and killed 36 h later. Transcriptional profiling by microarray and confirmation of selected mRNAs by real-time RT-PCR analyses was performed using total RNA from epithelial cells isolated from sections of the ampulla and isthmus collected from LP and FP treatment groups. Differentially expressed genes were subjected to gene ontology classification and bioinformatic pathway analyses.ResultsStatistical one-way ANOVA using Benjamini-hochberg multiple testing correction for false discovery rate (FDR) and pairwise comparison of epithelial cells in the ampulla of FP versus LP groups revealed 972 and 597 transcripts up- and down-regulated, respectively (P < 0.05). Within epithelial cells of the isthmus in FP versus LP groups, 946 and 817 transcripts were up- and down-regulated, respectively (P < 0.05). Up-regulated genes from both ampulla and isthmus were found to be largely involved in cholesterol biosynthesis and cell cycle pathways, while down-regulated genes were found in numerous inflammatory response pathways.ConclusionsMicroarray-based transcriptional profiling revealed phase of the cycle-dependent changes in the expression of mRNA within the epithelium of the oviducts’ ampulla and isthmus.Electronic supplementary materialThe online version of this article (doi:10.1186/s12958-015-0077-1) contains supplementary material, which is available to authorized users.

Highlights

Reproductive success depends on a functional oviduct for gamete storage, maturation, fertilization, and early embryonic development
Real-time RT- PCR analysis of selected transcripts The effect of stage of the estrous cycle and tissue on the expression of messenger RNA (mRNA) for NTS, binder of sperm 3 (BSP3), lactate dehydrogenase A (LDHA), cyclin D2 (CCND2), early growth response 1 (EGR1) and HSD17B7 was performed by real-time RTPCR
The expression of mRNA for NTS was found to increase by 32.4- and 150-fold within epithelial cells of the ampulla, and 21.6- and 88-fold within the epithelial cells of the isthmus, in

Summary

Introduction

Reproductive success depends on a functional oviduct for gamete storage, maturation, fertilization, and early embryonic development. The ovarian-derived steroids estrogen and progesterone are key regulators of oviductal function. Reproductive success depends on a functional oviduct for gamete storage and maturation, fertilization, and early embryonic development. Ovarian-derived steroids are well known regulators of oviductal function. Both estrogen and the progesterone receptors are abundant in the bovine oviducts’ mucosal epithelium [1,2,3,4,5,6], yet our understanding of how the steroidal environment affects the ability of the oviduct to function remains only partially understood. Maturation of gametes and breeding will occur in an estrogen dominant environment, fertilization and early cleavage after the steroidogenic shift and later stages of cleavage and formation of the morula occur within an oviduct exposed to increasing concentrations of circulating progesterone. Increasing our understanding of the steroidal control of oviductal function is critical to the design and implementation of interventions used to manage breeding and the establishment of a pregnancy.

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