Manipulation of bicarbonate concentration in sperm capacitation media improves in vitro fertilisation output in porcine species

作     者：Cristina Soriano-úbeda Jon Romero-Aguirregomezcorta Carmen Matás Pablo E.Visconti Francisco A.García-Vázquez 

作者机构：Department of PhysiologyFaculty of Veterinary ScienceInternational Excellence Campus for Higher Education and Research“Campus Mare Nostrum”University of Murcia30100 MurciaSpain Institute for Biomedical Research of Murcia(IMIB-Arrixaca)MurciaSpain Department of PhysiologyFaculty of Medicine and NursingUniversity of the Basque Country UPV/EHULeioaBizkaiaSpain.4Department of Veterinary and Animal SciencesUniversity of MassachusettsAmherstMAUSA. 

出 版 物：《Journal of Animal Science and Biotechnology》 (畜牧与生物技术杂志（英文版）)

年 卷 期：2019年第10卷第2期

页      面：375-389页

核心收录：

学科分类：0710[理学-生物学] 0831[工学-生物医学工程（可授工学、理学、医学学位）] 0832[工学-食品科学与工程（可授工学、农学学位）] 1004[医学-公共卫生与预防医学(可授医学、理学学位)] 0905[农学-畜牧学] 09[农学] 0906[农学-兽医学] 0703[理学-化学] 0836[工学-生物工程] 

基　　金：supported by the Spanish Ministry of Economy and Competitiveness(MINECO) the European Regional Development Fund(FEDER),Grants AGL2012–40180-C03–01-02 and AGL2015–66341-R) Fundación Séneca(20040/GERM/16) by a grant R01-HD-038082(to P.E.V.)from the National Institutes of Health(NIH) USA 

主　　题：Adcy10 Boar Monospermy PKA 

摘      要：Background: The in vivo concentration of bicarbonate(HCO3-), one of the essential sperm capacitating effectors,varies greatly in the different environments sperm go through from cauda epididymis to the fertilisation site. On the contrary, porcine in vitro sperm capacitation and fertilisation media usually contains a standard concentration of25 mmol/L, and one of the main problems presented is the unacceptable high incidence of polyspermy. This work hypothesised that by modifying the HCO3-concentration of the medium, the output of in vitro sperm capacitation and fertilisation could be increased.Results: Once exposed to the capacitation medium, the intracellular pH(pHi) of spermatozoa increased immediately even at low concentrations of HCO3-, but only extracellular concentrations of and above 15 mmol/L increased the substrates protein kinase A phosphorylation(pPKAs). Although with a significant delay, 15 mmol/L of HCO3-stimulated sperm linear motility and increased other late events in capacitation such as tyrosine phosphorylation(Tyr-P) to levels similar to those obtained with 25 mmol/L. This information allowed the establishment of a new in vitro fertilisation(IVF)system based on the optimization of HCO3-concentration to 15 mmol/L, which led to a 25.3% increment of the viable zygotes(8.6% in the standard system vs. 33.9%).Conclusions: Optimising HCO3-concentrations allows for establishing an IVF method that significantly reduced porcine polyspermy and increased the production of viable zygotes. A concentration of 15 mmol/L of HCO3-in the medium is sufficient to trigger the in vitro sperm capacitation and increase the fertilisation efficiency in porcine.