Arachidonic acid triggers [Ca2+]i increases in rat round spermatids by a likely GPR activation, ERK signalling and ER/acidic compartments Ca2+ release

PLoS One. 2017 Feb 13;12(2):e0172128. doi: 10.1371/journal.pone.0172128. eCollection 2017.

Abstract

Arachidonic acid (AA), a compound secreted by Sertoli cells (SC) in a FSH-dependent manner, is able to induce the release of Ca2+ from internal stores in round spermatids and pachytene spermatocytes. In this study, the possible site(s) of action of AA in round spermatids, the signalling pathways associated and the intracellular Ca2+ stores targeted by AA-induced signalling were pharmacologically characterized by measuring intracellular Ca2+ using fluorescent Ca2+ probes. Our results suggest that AA acts by interacting with a fatty acid G protein coupled receptor, initiating a G protein signalling cascade that may involve PLA2 and ERK activation, which in turn opens intracellular ryanodine-sensitive channels as well as NAADP-sensitive channels in acidic intracellular Ca2+ stores. The results presented here also suggest that AMPK and PKA modulate this AA-induced Ca2+ release from intracellular Ca2+ stores in round spermatids. We propose that unsaturated free fatty acid lipid signalling in the seminiferous tubule is a novel regulatory component of rat spermatogenesis.

MeSH terms

  • Animals
  • Arachidonic Acid / pharmacology*
  • Calcium / metabolism*
  • Cells, Cultured
  • Dose-Response Relationship, Drug
  • Endoplasmic Reticulum / drug effects*
  • Endoplasmic Reticulum / metabolism
  • Endosomes / drug effects
  • Endosomes / metabolism
  • Kinetics
  • MAP Kinase Signaling System / drug effects*
  • Male
  • Microscopy, Confocal
  • NADP / analogs & derivatives
  • NADP / metabolism
  • Phospholipases A2 / metabolism
  • Rats, Sprague-Dawley
  • Receptors, G-Protein-Coupled / agonists*
  • Receptors, G-Protein-Coupled / metabolism
  • Salicylates / pharmacology
  • Sesterterpenes / pharmacology
  • Spermatids / cytology
  • Spermatids / drug effects*
  • Spermatids / metabolism
  • Testis / cytology
  • Testis / drug effects
  • Testis / metabolism

Substances

  • Ffar4 protein, rat
  • Receptors, G-Protein-Coupled
  • Salicylates
  • Sesterterpenes
  • grifolic acid methyl ether
  • nicotinic acid adenine dinucleotide phosphate acetoxymethyl ester
  • Arachidonic Acid
  • NADP
  • Phospholipases A2
  • Calcium

Grants and funding

This work was supported by Fondecyt grant numbers 1110267 and 1140758; and PUCV grant number 125.754/2011 125792/2014 to JGR; and Consejo Nacional de Ciencia y Tecnología (CONACyT-Mexico) (128566 to CT), Dirección General de Asuntos del Personal Académico/ Universidad Nacional Autónoma de México (DGAPA/ UNAM) (IN202212-3 to CT) and The Alexander von Humboldt Foundation (to CT). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.