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Manipulation of bicarbonate concentration in sperm capacitation media improvesin vitro fertilisation output in porcine species
BACKGROUND: The in vivo concentration of bicarbonate (HCO(3)(−)), 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 m...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6410524/ https://www.ncbi.nlm.nih.gov/pubmed/30899459 http://dx.doi.org/10.1186/s40104-019-0324-y |
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author | Soriano-Úbeda, Cristina Romero-Aguirregomezcorta, Jon Matás, Carmen Visconti, Pablo E. García-Vázquez, Francisco A. |
author_facet | Soriano-Úbeda, Cristina Romero-Aguirregomezcorta, Jon Matás, Carmen Visconti, Pablo E. García-Vázquez, Francisco A. |
author_sort | Soriano-Úbeda, Cristina |
collection | PubMed |
description | BACKGROUND: The in vivo concentration of bicarbonate (HCO(3)(−)), 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 of 25 mmol/L, and one of the main problems presented is the unacceptable high incidence of polyspermy. This work hypothesised that by modifying the HCO(3)(−) 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 (pH(i)) of spermatozoa increased immediately even at low concentrations of HCO(3)(−), 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 HCO(3)(−) 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 HCO(3)(−) 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 HCO(3)(−) 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 HCO(3)(−) in the medium is sufficient to trigger the in vitro sperm capacitation and increase the fertilisation efficiency in porcine. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s40104-019-0324-y) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-6410524 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-64105242019-03-21 Manipulation of bicarbonate concentration in sperm capacitation media improvesin vitro fertilisation output in porcine species Soriano-Úbeda, Cristina Romero-Aguirregomezcorta, Jon Matás, Carmen Visconti, Pablo E. García-Vázquez, Francisco A. J Anim Sci Biotechnol Research BACKGROUND: The in vivo concentration of bicarbonate (HCO(3)(−)), 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 of 25 mmol/L, and one of the main problems presented is the unacceptable high incidence of polyspermy. This work hypothesised that by modifying the HCO(3)(−) 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 (pH(i)) of spermatozoa increased immediately even at low concentrations of HCO(3)(−), 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 HCO(3)(−) 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 HCO(3)(−) 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 HCO(3)(−) 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 HCO(3)(−) in the medium is sufficient to trigger the in vitro sperm capacitation and increase the fertilisation efficiency in porcine. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s40104-019-0324-y) contains supplementary material, which is available to authorized users. BioMed Central 2019-03-11 /pmc/articles/PMC6410524/ /pubmed/30899459 http://dx.doi.org/10.1186/s40104-019-0324-y Text en © The Author(s). 2019 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Soriano-Úbeda, Cristina Romero-Aguirregomezcorta, Jon Matás, Carmen Visconti, Pablo E. García-Vázquez, Francisco A. Manipulation of bicarbonate concentration in sperm capacitation media improvesin vitro fertilisation output in porcine species |
title | Manipulation of bicarbonate concentration in sperm capacitation media improvesin vitro fertilisation output in porcine species |
title_full | Manipulation of bicarbonate concentration in sperm capacitation media improvesin vitro fertilisation output in porcine species |
title_fullStr | Manipulation of bicarbonate concentration in sperm capacitation media improvesin vitro fertilisation output in porcine species |
title_full_unstemmed | Manipulation of bicarbonate concentration in sperm capacitation media improvesin vitro fertilisation output in porcine species |
title_short | Manipulation of bicarbonate concentration in sperm capacitation media improvesin vitro fertilisation output in porcine species |
title_sort | manipulation of bicarbonate concentration in sperm capacitation media improvesin vitro fertilisation output in porcine species |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6410524/ https://www.ncbi.nlm.nih.gov/pubmed/30899459 http://dx.doi.org/10.1186/s40104-019-0324-y |
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