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Human Sperm Capacitation Involves the Regulation of the Tyr-Phosphorylation Level of the Anion Exchanger 1 (AE1)

Bicarbonate uptake is one of the early steps of capacitation, but the identification of proteins regulating anion fluxes remains elusive. The aim of this study is to investigate the role of sperm solute carrier 4 (SLC4) A1 (spAE1) in the capacitation process. The expression, location, and tyrosine-p...

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Autores principales: Donà, Gabriella, Tibaldi, Elena, Andrisani, Alessandra, Ambrosini, Guido, Sabbadin, Chiara, Pagano, Mario Angelo, Brunati, Anna Maria, Armanini, Decio, Ragazzi, Eugenio, Bordin, Luciana
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
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Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7311965/
https://www.ncbi.nlm.nih.gov/pubmed/32517126
http://dx.doi.org/10.3390/ijms21114063
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author Donà, Gabriella
Tibaldi, Elena
Andrisani, Alessandra
Ambrosini, Guido
Sabbadin, Chiara
Pagano, Mario Angelo
Brunati, Anna Maria
Armanini, Decio
Ragazzi, Eugenio
Bordin, Luciana
author_facet Donà, Gabriella
Tibaldi, Elena
Andrisani, Alessandra
Ambrosini, Guido
Sabbadin, Chiara
Pagano, Mario Angelo
Brunati, Anna Maria
Armanini, Decio
Ragazzi, Eugenio
Bordin, Luciana
author_sort Donà, Gabriella
collection PubMed
description Bicarbonate uptake is one of the early steps of capacitation, but the identification of proteins regulating anion fluxes remains elusive. The aim of this study is to investigate the role of sperm solute carrier 4 (SLC4) A1 (spAE1) in the capacitation process. The expression, location, and tyrosine-phosphorylation (Tyr-P) level of spAE1 were assessed. Thereby, it was found that 4,4′-Diisothiocyano-2,2′-stilbenedisulfonic acid (DIDS), an SLC4 family channel blocker, inhibited capacitation in a dose-dependent manner by decreasing acrosome reaction (ARC% 24.5 ± 3.3 vs. 64.9 ± 4.3, p < 0.05) and increasing the percentage of not viable cells (NVC%), comparable to the inhibition by I-172, a cystic fibrosis transmembrane conductance regulator (CFTR) blocker (AR% 30.5 ± 4.4 and NVC% 18.6 ± 2.2). When used in combination, a synergistic inhibitory effect was observed with a remarkable increase of the percentage of NVC (45.3 ± 4.1, p < 0.001). spAE1 was identified in sperm membrane as a substrate for Tyr-protein kinases Lyn and Syk, which were identified as both soluble and membrane-bound pools. spAE1-Tyr-P level increased in the apical region of sperm under capacitating conditions and was negatively affected by I-172 or DIDS, and, to a far greater extent, by a combination of both. In conclusion, we demonstrated that spAE1 is expressed in sperm membranes and it is phosphorylated by Syk, but above all by Lyn on Tyr359, which are involved in sperm viability and capacitation.
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spelling pubmed-73119652020-06-25 Human Sperm Capacitation Involves the Regulation of the Tyr-Phosphorylation Level of the Anion Exchanger 1 (AE1) Donà, Gabriella Tibaldi, Elena Andrisani, Alessandra Ambrosini, Guido Sabbadin, Chiara Pagano, Mario Angelo Brunati, Anna Maria Armanini, Decio Ragazzi, Eugenio Bordin, Luciana Int J Mol Sci Article Bicarbonate uptake is one of the early steps of capacitation, but the identification of proteins regulating anion fluxes remains elusive. The aim of this study is to investigate the role of sperm solute carrier 4 (SLC4) A1 (spAE1) in the capacitation process. The expression, location, and tyrosine-phosphorylation (Tyr-P) level of spAE1 were assessed. Thereby, it was found that 4,4′-Diisothiocyano-2,2′-stilbenedisulfonic acid (DIDS), an SLC4 family channel blocker, inhibited capacitation in a dose-dependent manner by decreasing acrosome reaction (ARC% 24.5 ± 3.3 vs. 64.9 ± 4.3, p < 0.05) and increasing the percentage of not viable cells (NVC%), comparable to the inhibition by I-172, a cystic fibrosis transmembrane conductance regulator (CFTR) blocker (AR% 30.5 ± 4.4 and NVC% 18.6 ± 2.2). When used in combination, a synergistic inhibitory effect was observed with a remarkable increase of the percentage of NVC (45.3 ± 4.1, p < 0.001). spAE1 was identified in sperm membrane as a substrate for Tyr-protein kinases Lyn and Syk, which were identified as both soluble and membrane-bound pools. spAE1-Tyr-P level increased in the apical region of sperm under capacitating conditions and was negatively affected by I-172 or DIDS, and, to a far greater extent, by a combination of both. In conclusion, we demonstrated that spAE1 is expressed in sperm membranes and it is phosphorylated by Syk, but above all by Lyn on Tyr359, which are involved in sperm viability and capacitation. MDPI 2020-06-05 /pmc/articles/PMC7311965/ /pubmed/32517126 http://dx.doi.org/10.3390/ijms21114063 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Donà, Gabriella
Tibaldi, Elena
Andrisani, Alessandra
Ambrosini, Guido
Sabbadin, Chiara
Pagano, Mario Angelo
Brunati, Anna Maria
Armanini, Decio
Ragazzi, Eugenio
Bordin, Luciana
Human Sperm Capacitation Involves the Regulation of the Tyr-Phosphorylation Level of the Anion Exchanger 1 (AE1)
title Human Sperm Capacitation Involves the Regulation of the Tyr-Phosphorylation Level of the Anion Exchanger 1 (AE1)
title_full Human Sperm Capacitation Involves the Regulation of the Tyr-Phosphorylation Level of the Anion Exchanger 1 (AE1)
title_fullStr Human Sperm Capacitation Involves the Regulation of the Tyr-Phosphorylation Level of the Anion Exchanger 1 (AE1)
title_full_unstemmed Human Sperm Capacitation Involves the Regulation of the Tyr-Phosphorylation Level of the Anion Exchanger 1 (AE1)
title_short Human Sperm Capacitation Involves the Regulation of the Tyr-Phosphorylation Level of the Anion Exchanger 1 (AE1)
title_sort human sperm capacitation involves the regulation of the tyr-phosphorylation level of the anion exchanger 1 (ae1)
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7311965/
https://www.ncbi.nlm.nih.gov/pubmed/32517126
http://dx.doi.org/10.3390/ijms21114063
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