RESEARCH ARTICLE
The role of cGMP as a mediator of lipolysis in bovine oocytes and its effects on embryo development and cryopreservation Ka´tia R. L. Schwarz1*, Fernanda C. de Castro1, Letı´cia Schefer1, Ramon C. Botigelli2, Daniela M. Paschoal1, Hugo Fernandes1, Cla´udia L. V. Leal1*
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1 Universidade de São Paulo (USP), Faculdade de Zootecnia e Engenharia de Alimentos (FZEA), Departamento de Medicina Veterina´ria, Pirassununga, São Paulo, Brasil, 2 Universidade Estadual Paulista (Unesp), Instituto de Biociências de Botucatu (IBB), Departamento de Farmacologia, Botucatu, São Paulo, Brasil *
[email protected] (CLVL);
[email protected] (KRLS)
Abstract
OPEN ACCESS Citation: Schwarz KRL, de Castro FC, Schefer L, Botigelli RC, Paschoal DM, Fernandes H, et al. (2018) The role of cGMP as a mediator of lipolysis in bovine oocytes and its effects on embryo development and cryopreservation. PLoS ONE 13(1): e0191023. https://doi.org/10.1371/journal. pone.0191023 Editor: Deborah A. Altomare, University of Central Florida, UNITED STATES Received: February 8, 2017 Accepted: December 27, 2017 Published: January 23, 2018 Copyright: © 2018 Schwarz et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Data Availability Statement: All relevant data are within the paper. Funding: This work was supported by Grant numbers: 2013/15374-3; 2015/20379-0, www. fapesp.br. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
This study aimed to determine the influence of cyclic guanosine 3’5’-monophosphate (cGMP) and cGMP-dependent kinase (PKG) during in vitro maturation (IVM) on lipolysisrelated parameters in bovine cumulus-oocyte complexes (COCs), and on embryo development and cryosurvival. COCs were matured with cGMP/PKG modulators and assessed for metaphase II rates (MII), cGMP levels, lipid content in oocytes (OO), transcript abundance for genes involved in lipolysis (ATGL) and lipid droplets (PLIN2) in cumulus cells (CC) and OO, and presence of phosphorylated (active) hormone sensitive lipase (HSLser563) in OO. Embryo development, lipid contents and survival to vitrification were also assessed. Phosphodiesterase 5 inhibition (PDE5; cGMP-hydrolyzing enzyme) with 10-5M sildenafil (SDF) during 24 h IVM increased cGMP in COCs (56.9 vs 9.5 fMol/COC in untreated controls, p
