Cargando…

Modulatory effect of MG-132 proteasomal inhibition on boar sperm motility during in vitro capacitation

A series of biochemical and biophysical changes during sperm capacitation initiates various signaling pathways related to protein phosphorylation leading to sperm hyperactivation, simultaneously with the regulation of proteasomal activity responsible for protein degradation and turnover. Our study a...

Descripción completa

Detalles Bibliográficos
Autores principales: Hackerova, Lenka, Klusackova, Barbora, Zigo, Michal, Zelenkova, Natalie, Havlikova, Katerina, Krejcirova, Romana, Sedmikova, Marketa, Sutovsky, Peter, Komrskova, Katerina, Postlerova, Pavla, Simonik, Ondrej
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10077870/
https://www.ncbi.nlm.nih.gov/pubmed/37035827
http://dx.doi.org/10.3389/fvets.2023.1116891
_version_ 1785020398487732224
author Hackerova, Lenka
Klusackova, Barbora
Zigo, Michal
Zelenkova, Natalie
Havlikova, Katerina
Krejcirova, Romana
Sedmikova, Marketa
Sutovsky, Peter
Komrskova, Katerina
Postlerova, Pavla
Simonik, Ondrej
author_facet Hackerova, Lenka
Klusackova, Barbora
Zigo, Michal
Zelenkova, Natalie
Havlikova, Katerina
Krejcirova, Romana
Sedmikova, Marketa
Sutovsky, Peter
Komrskova, Katerina
Postlerova, Pavla
Simonik, Ondrej
author_sort Hackerova, Lenka
collection PubMed
description A series of biochemical and biophysical changes during sperm capacitation initiates various signaling pathways related to protein phosphorylation leading to sperm hyperactivation, simultaneously with the regulation of proteasomal activity responsible for protein degradation and turnover. Our study aimed to unveil the role of the proteasome in the regulation of boar sperm motility, hyperactivated status, tyrosine phosphorylation, and total protein ubiquitination. The proteolytic activity of the 20S proteasomal core was inhibited by MG-132 in concentrations of 10, 25, 50, and 100 μM; and monitored parameters were analyzed every hour during 3 h of in vitro capacitation (IVC). Sperm motility and kinematic parameters were analyzed by Computer Assisted Sperm Analysis (CASA) during IVC, showing a significant, negative, dose-dependent effect of MG-132 on total and progressive sperm motility (TMOT, PMOT, respectively). Furthermore, proteasomal inhibition by 50 and 100 μM MG-132 had a negative impact on velocity-based kinematic sperm parameters (VSL, VAP, and VCL). Parameters related to the progressivity of sperm movement (LIN, STR) and ALH were the most affected by the highest inhibitor concentration (100 μM). Cluster analysis revealed that the strongest proteasome-inhibiting treatment had a significant effect (p ≤ 0.05) on the hyperactivated sperm subpopulation. The flow cytometric viability results proved that reduced TMOT and PMOT were not caused by disruption of the integrity of the plasma membrane. Neither the protein tyrosine phosphorylation profile changes nor the accumulation of protein ubiquitination was observed during the course of capacitation under proteasome inhibition. In conclusion, inhibition of the proteasome reduced the ability of spermatozoa to undergo hyperactivation; however, there was no significant effect on the level of protein tyrosine phosphorylation and accumulation of ubiquitinated proteins. These effects might be due to the presence of compensatory mechanisms or the alteration of various ubiquitin-proteasome system-regulated pathways.
format Online
Article
Text
id pubmed-10077870
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-100778702023-04-07 Modulatory effect of MG-132 proteasomal inhibition on boar sperm motility during in vitro capacitation Hackerova, Lenka Klusackova, Barbora Zigo, Michal Zelenkova, Natalie Havlikova, Katerina Krejcirova, Romana Sedmikova, Marketa Sutovsky, Peter Komrskova, Katerina Postlerova, Pavla Simonik, Ondrej Front Vet Sci Veterinary Science A series of biochemical and biophysical changes during sperm capacitation initiates various signaling pathways related to protein phosphorylation leading to sperm hyperactivation, simultaneously with the regulation of proteasomal activity responsible for protein degradation and turnover. Our study aimed to unveil the role of the proteasome in the regulation of boar sperm motility, hyperactivated status, tyrosine phosphorylation, and total protein ubiquitination. The proteolytic activity of the 20S proteasomal core was inhibited by MG-132 in concentrations of 10, 25, 50, and 100 μM; and monitored parameters were analyzed every hour during 3 h of in vitro capacitation (IVC). Sperm motility and kinematic parameters were analyzed by Computer Assisted Sperm Analysis (CASA) during IVC, showing a significant, negative, dose-dependent effect of MG-132 on total and progressive sperm motility (TMOT, PMOT, respectively). Furthermore, proteasomal inhibition by 50 and 100 μM MG-132 had a negative impact on velocity-based kinematic sperm parameters (VSL, VAP, and VCL). Parameters related to the progressivity of sperm movement (LIN, STR) and ALH were the most affected by the highest inhibitor concentration (100 μM). Cluster analysis revealed that the strongest proteasome-inhibiting treatment had a significant effect (p ≤ 0.05) on the hyperactivated sperm subpopulation. The flow cytometric viability results proved that reduced TMOT and PMOT were not caused by disruption of the integrity of the plasma membrane. Neither the protein tyrosine phosphorylation profile changes nor the accumulation of protein ubiquitination was observed during the course of capacitation under proteasome inhibition. In conclusion, inhibition of the proteasome reduced the ability of spermatozoa to undergo hyperactivation; however, there was no significant effect on the level of protein tyrosine phosphorylation and accumulation of ubiquitinated proteins. These effects might be due to the presence of compensatory mechanisms or the alteration of various ubiquitin-proteasome system-regulated pathways. Frontiers Media S.A. 2023-03-23 /pmc/articles/PMC10077870/ /pubmed/37035827 http://dx.doi.org/10.3389/fvets.2023.1116891 Text en Copyright © 2023 Hackerova, Klusackova, Zigo, Zelenkova, Havlikova, Krejcirova, Sedmikova, Sutovsky, Komrskova, Postlerova and Simonik. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Veterinary Science
Hackerova, Lenka
Klusackova, Barbora
Zigo, Michal
Zelenkova, Natalie
Havlikova, Katerina
Krejcirova, Romana
Sedmikova, Marketa
Sutovsky, Peter
Komrskova, Katerina
Postlerova, Pavla
Simonik, Ondrej
Modulatory effect of MG-132 proteasomal inhibition on boar sperm motility during in vitro capacitation
title Modulatory effect of MG-132 proteasomal inhibition on boar sperm motility during in vitro capacitation
title_full Modulatory effect of MG-132 proteasomal inhibition on boar sperm motility during in vitro capacitation
title_fullStr Modulatory effect of MG-132 proteasomal inhibition on boar sperm motility during in vitro capacitation
title_full_unstemmed Modulatory effect of MG-132 proteasomal inhibition on boar sperm motility during in vitro capacitation
title_short Modulatory effect of MG-132 proteasomal inhibition on boar sperm motility during in vitro capacitation
title_sort modulatory effect of mg-132 proteasomal inhibition on boar sperm motility during in vitro capacitation
topic Veterinary Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10077870/
https://www.ncbi.nlm.nih.gov/pubmed/37035827
http://dx.doi.org/10.3389/fvets.2023.1116891
work_keys_str_mv AT hackerovalenka modulatoryeffectofmg132proteasomalinhibitiononboarspermmotilityduringinvitrocapacitation
AT klusackovabarbora modulatoryeffectofmg132proteasomalinhibitiononboarspermmotilityduringinvitrocapacitation
AT zigomichal modulatoryeffectofmg132proteasomalinhibitiononboarspermmotilityduringinvitrocapacitation
AT zelenkovanatalie modulatoryeffectofmg132proteasomalinhibitiononboarspermmotilityduringinvitrocapacitation
AT havlikovakaterina modulatoryeffectofmg132proteasomalinhibitiononboarspermmotilityduringinvitrocapacitation
AT krejcirovaromana modulatoryeffectofmg132proteasomalinhibitiononboarspermmotilityduringinvitrocapacitation
AT sedmikovamarketa modulatoryeffectofmg132proteasomalinhibitiononboarspermmotilityduringinvitrocapacitation
AT sutovskypeter modulatoryeffectofmg132proteasomalinhibitiononboarspermmotilityduringinvitrocapacitation
AT komrskovakaterina modulatoryeffectofmg132proteasomalinhibitiononboarspermmotilityduringinvitrocapacitation
AT postlerovapavla modulatoryeffectofmg132proteasomalinhibitiononboarspermmotilityduringinvitrocapacitation
AT simonikondrej modulatoryeffectofmg132proteasomalinhibitiononboarspermmotilityduringinvitrocapacitation