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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MDPI AG
2020-06-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/21/11/4063 |
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| author | Gabriella Donà Elena Tibaldi Alessandra Andrisani Guido Ambrosini Chiara Sabbadin Mario Angelo Pagano Anna Maria Brunati Decio Armanini Eugenio Ragazzi Luciana Bordin |
| author_facet | Gabriella Donà Elena Tibaldi Alessandra Andrisani Guido Ambrosini Chiara Sabbadin Mario Angelo Pagano Anna Maria Brunati Decio Armanini Eugenio Ragazzi Luciana Bordin |
| author_sort | Gabriella Donà |
| collection | DOAJ |
| 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, <i>p</i> < 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, <i>p</i> < 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. |
| first_indexed | 2024-03-10T19:20:59Z |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T19:20:59Z |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-86b47112f1e147b19dc8e99cda1a0bdf2023-11-20T03:01:09ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672020-06-012111406310.3390/ijms21114063Human Sperm Capacitation Involves the Regulation of the Tyr-Phosphorylation Level of the Anion Exchanger 1 (AE1)Gabriella Donà0Elena Tibaldi1Alessandra Andrisani2Guido Ambrosini3Chiara Sabbadin4Mario Angelo Pagano5Anna Maria Brunati6Decio Armanini7Eugenio Ragazzi8Luciana Bordin9Department of Molecular Medicine-Biological Chemistry, University of Padua, 35131 Padua, ItalyDepartment of Molecular Medicine-Biological Chemistry, University of Padua, 35131 Padua, ItalyDepartment of Women’s and Children’s Health, University of Padua, 35131 Padua, ItalyDepartment of Women’s and Children’s Health, University of Padua, 35131 Padua, ItalyDepartment of Medicine–Endocrinology, University of Padua, 35131 Padua, ItalyDepartment of Molecular Medicine-Biological Chemistry, University of Padua, 35131 Padua, ItalyDepartment of Molecular Medicine-Biological Chemistry, University of Padua, 35131 Padua, ItalyDepartment of Medicine–Endocrinology, University of Padua, 35131 Padua, ItalyDepartment of Pharmaceutical and Pharmacological Sciences, University of Padua, 35131 Padua, ItalyDepartment of Molecular Medicine-Biological Chemistry, University of Padua, 35131 Padua, ItalyBicarbonate 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, <i>p</i> < 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, <i>p</i> < 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.https://www.mdpi.com/1422-0067/21/11/4063spermSLC4A1AE1Tyr-phosphorylationcapacitationacrosome reaction |
| spellingShingle | Gabriella Donà Elena Tibaldi Alessandra Andrisani Guido Ambrosini Chiara Sabbadin Mario Angelo Pagano Anna Maria Brunati Decio Armanini Eugenio Ragazzi Luciana Bordin Human Sperm Capacitation Involves the Regulation of the Tyr-Phosphorylation Level of the Anion Exchanger 1 (AE1) International Journal of Molecular Sciences sperm SLC4A1 AE1 Tyr-phosphorylation capacitation acrosome reaction |
| 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 | sperm SLC4A1 AE1 Tyr-phosphorylation capacitation acrosome reaction |
| url | https://www.mdpi.com/1422-0067/21/11/4063 |
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