The prostaglandin E2 receptor PTGER2 and prostaglandin F receptor PTGFR mediate oviductal glycoprotein 1 expression in bovine oviductal epithelial cells

J Reprod Dev. 2018 Apr 13;64(2):101-108. doi: 10.1262/jrd.2017-076. Epub 2017 Dec 23.

Abstract

Oviductal glycoprotein 1 (OVGP1), an oviductin, is involved in the maintenance of sperm viability and motility and contributes to sperm capacitation in the oviduct. In this study, the regulatory effects exerted by prostaglandin E2 (PGE2) and F (PGF) on OVGP1 expression via their corresponding receptors in bovine oviductal epithelial cells (BOECs) were investigated. BOECs were cultured in vitro, and their expression of receptors of PGE2 (PTGER1, PTGER2, PTGER3, and PTGER4) and PGF (PTGFR) was measured using RT-qPCR. Ca2+ concentration was determined with a fluorescence-based method and cAMP was quantified by enzyme-linked immunosorbent assays to verify activation of PTGER2 and PTGFR by their corresponding agonists in these cells. OVGP1 mRNA and protein expression was measured using RT-qPCR and western blotting, respectively, following PTGER2 and PTGFR agonist-induced activation. PTGER1, PTGER2, PTGER4, and PTGFR were found to be present in BOECs; however, PTGER3 expression was not detected. OVGP1 expression was significantly promoted by 10-6 M butaprost (a PTGER2 agonist) and decreased by 10-6 M fluprostenol (a PTGFR agonist). In addition, 3 μM H-89 (a PKA inhibitor) and 3 μM U0126 (an ERK inhibitor) effectively inhibited PGE2-induced upregulation of OVGP1, and 5 μM chelerythrine chloride (a PKC inhibitor) and 3 μM U0126 negated OVGP1 downregulation by PGF. In conclusion, this study demonstrates that OVGP1 expression in BOECs is enhanced by PGE2 via PTGER2-cAMP-PKA signaling, and reduced by PGF through the PTGFR-Ca2+-PKC pathway.

Keywords: Oviduct; Oviductal glycoprotein 1 (OVGP1); PTGER2; PTGFR; Prostaglandin E2(PGE2); Prostaglandin F2α(PGF2α).

MeSH terms

  • Abattoirs
  • Alprostadil / analogs & derivatives
  • Alprostadil / pharmacology
  • Animals
  • Calcium Signaling / drug effects
  • Cattle
  • Cells, Cultured
  • Cyclic AMP / metabolism
  • Cyclic AMP-Dependent Protein Kinases / antagonists & inhibitors
  • Cyclic AMP-Dependent Protein Kinases / chemistry
  • Cyclic AMP-Dependent Protein Kinases / metabolism
  • Dinoprost / metabolism*
  • Dinoprostone / metabolism*
  • Female
  • Gene Expression Regulation* / drug effects
  • Glycoproteins / agonists
  • Glycoproteins / antagonists & inhibitors
  • Glycoproteins / genetics
  • Glycoproteins / metabolism*
  • Oviducts / cytology
  • Oviducts / drug effects
  • Oviducts / metabolism*
  • Prostaglandins F, Synthetic / pharmacology
  • Protein Isoforms / agonists
  • Protein Isoforms / metabolism
  • Protein Kinase C / antagonists & inhibitors
  • Protein Kinase C / chemistry
  • Protein Kinase C / metabolism
  • Protein Kinase Inhibitors / pharmacology
  • Receptors, Prostaglandin / agonists*
  • Receptors, Prostaglandin / metabolism
  • Receptors, Prostaglandin E, EP2 Subtype / agonists*
  • Receptors, Prostaglandin E, EP2 Subtype / metabolism

Substances

  • Glycoproteins
  • OVGP1 protein, Bos taurus
  • Prostaglandins F, Synthetic
  • Protein Isoforms
  • Protein Kinase Inhibitors
  • Receptors, Prostaglandin
  • Receptors, Prostaglandin E, EP2 Subtype
  • prostaglandin F2alpha receptor
  • fluprostenol
  • Dinoprost
  • Cyclic AMP
  • Cyclic AMP-Dependent Protein Kinases
  • Protein Kinase C
  • Alprostadil
  • butaprost
  • Dinoprostone