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

BACKGROUND: 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. 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. METHODS: Six 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 PGF(2α) (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. RESULTS: Statistical 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. CONCLUSIONS: Microarray-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 MATERIAL: The online version of this article (doi:10.1186/s12958-015-0077-1) contains supplementary material, which is available to authorized users

Bovine in vitro produced embryos induce modifications of gene expression profile of bovine oviduct epithelial cells in vitro

In vivo, the ovicluct provicles the optimal environment to allow the sucœssful clevelopment of the carly mammalian embryo. A molecular dialogue may take place between the cmbryo and its maternai milieu, moclulating the embryo microenvironment clnring its migration towm*ds the implantation site. ln vitro co-culture witb bovine ovicluct epithelial œlls (BOEC) bas been widely used to mimic this materna] environment and to study its effect on embryo development. The exact mechanisms of BOEC action on embryo development have not been l'ully elucidated yet. Therefore, the pUL*pose of this research was to evaluate the BOEC in vitro rcsponsiveness to embryos, accorcling to the regional origin of the oviduct cells (isthmus vs. ampulla). Oviducts ipsilateral to ovnlies with sign of recent ovulation were brought to the laboratory. Exp. l: Ampulla and istbmus regions were dissected, washed thoroughly in TCM199 and epithelial cells werc scrappecl out using a sterile slide. BOEC from Arnpulla (A-BOEC) or lsthmus (1-BOEC) were seeded separately in 4 weil NUNC plates and cultured to confluence (7 days) to be used for in vitro embryo development (IVD). Immature cumulus oocyte complexes were aspirated from slaughterhouse avaries. Zygotes produced by in vitro maturation and fcrtilization wcrc culturecl in SOF medium supplementecl with 10% FCS in the presence of A-BOEC, l-BOEC or without cells (control). Somc A- and I-BOEC wells were culturecl without embryos. At Day 8 p.i., RNA were extracted (Trizol). Exp. 2) Confluent BOEC in 4 well NUNC plates were stimulated by synthetic INFtau (0. l, 10 and lOO ng/mL) for 6 or 24 hours and RNA were exa*acted (Trizol). Ali RNA srm1ples were treated with DNAsc and RT was petformed (MMLV RT kit). The leve! of expression of some known ovicluct exprcssecl genes (GPX4, C3, OVGP), as well as some genes related to IFN signaling (STATl, !FIT5, ISG15, OASl, IFITMl, MXl, OASl, USP18), wcre cvaluatcd by RT-qPCR. Data were analyzed by Mann Whitney non parmnetric test using PRISM 5 software. TI!C relative abun dance of ali IFNt related genes was significantly upregttlatecl whcn epithelial cells (either A-BOEC or I BOEC) were exposecl to embryos duling 8 days in vitro. The IFNI stimulation reproduced tllis embryo clfect, whatever the concentration tmcl duration of treatment. Furthennore, a regional difference in exprcssionlcvcl was found for OVGP (higher in I-BOEC, p<0.05) and C3 (bighcr in A-BOEC, p<0.05), without effect of the presence of embryos. GPX4 mRNA abundanœ was not significantly different among the culture conditions tested. In conclusion, these data confirm the specializatiou of ovicluct regions and show the ability of oviduet œlls to respond to emlHyo signaling. at !east ù1rough lFN-Iike pathway in our in vitro mode!, by modulating gene expression profile, thus suppo1ting the existence of a rerdialog betwcen carly embryo tmd oviduct

Positive effects of bovine oviduct epithelial cells on goat embryos development and cryosurvival in vitro

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Regulation of Cell Fate Decisions in Early Mammalian Embryos

Early embryogenesis is characterized by the segregation of cell lineages that fulfill critical roles in the establishment of pregnancy and development of the fetus. The formation of the blastocyst marks the emergence of extraembryonic precursors, needed for implantation, and of pluripotent cells, which differentiate toward the major lineages of the adult organism. The coordinated emergence of these cell types shows that these processes are broadly conserved in mammals. However, developmental heterochrony and changes in gene regulatory networks highlight unique evolutionary adaptations that may explain the diversity in placentation and in the mechanisms controlling pluripotency in mammals. The incorporation of new technologies, including single-cell omics, imaging, and gene editing, is instrumental for comparative embryology. Broadening the knowledge of mammalian embryology will provide new insights into the mechanisms driving evolution and development. This knowledge can be readily translated into biomedical and biotechnological applications in humans and livestock, respectively

Transcriptional reconfiguration of rabbit iPS cells with Krüppel-like factors confers chimeric competency

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Transcriptional reconfiguration of rabbit iPS cells with Krüppel-like factors confers chimeric competency

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Identification of biallelic germline variants of SRP68 in a sporadic case with severe congenital neutropenia

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