Impact of Cryopreservation on Motile Subpopulations and Tyrosine-Phosphorylated Regions of Ram Spermatozoa during Capacitating Conditions

The heterogeneous nature of ejaculates highlights the relevance of studying the behavior of different sperm subpopulations. Changes in sperm motility and the increase in tyrosine phosphorylation are key events that usually occur during capacitation and can be modified by the cryopreservation process...

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Autores principales:	Patricia Peris-Frau, Irene Sánchez-Ajofrín, Alicia Martín Maestro, Carolina Maside, Daniela Alejandra Medina-Chávez, Olga García-Álvarez, María del Rocío Fernández-Santos, Vidal Montoro, José Julián Garde, Manuel Ramón, Ana Josefa Soler
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	computer-assisted sperm analysis cytometry subpopulations sperm capacitation sperm cryopreservation tyrosine phosphorylation Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/f6abda8580574ed7a7a8ca969d7a03a3
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spelling	oai:doaj.org-article:f6abda8580574ed7a7a8ca969d7a03a32021-11-25T16:48:10ZImpact of Cryopreservation on Motile Subpopulations and Tyrosine-Phosphorylated Regions of Ram Spermatozoa during Capacitating Conditions10.3390/biology101112132079-7737https://doaj.org/article/f6abda8580574ed7a7a8ca969d7a03a32021-11-01T00:00:00Zhttps://www.mdpi.com/2079-7737/10/11/1213https://doaj.org/toc/2079-7737The heterogeneous nature of ejaculates highlights the relevance of studying the behavior of different sperm subpopulations. Changes in sperm motility and the increase in tyrosine phosphorylation are key events that usually occur during capacitation and can be modified by the cryopreservation process. However, the relationship between both events remains poorly defined throughout capacitation in the different sperm subpopulations. Fresh and frozen-thawed spermatozoa were incubated in capacitating (CAP) and non-capacitating (NC) media up to 240 min. Sperm kinematics, tyrosine phosphorylation and mitochondrial activity were measured by the CASA system and imaging flow cytometry. Four motile sperm subpopulations (SP) were identified in fresh and frozen-thawed ram semen after the cluster analysis. Incubation under CAP conditions over time led to greater changes in the percentage of spermatozoa included in each subpopulation compared to NC conditions, being different between fresh and frozen-thawed spermatozoa. The SP1, characterized by slow spermatozoa, progressively increased after 15 min in frozen-thawed samples incubated in both media but not in fresh ones. The SP4, characterized by fast and non-linear spermatozoa, showed a marked increase during CAP, but not under NC conditions, occurring more rapidly in frozen-thawed spermatozoa. This subpopulation (SP4) was also the only one positively and strongly correlated with mitochondrial activity and all phosphorylated sperm regions during capacitation, either in fresh or frozen-thawed samples. Our results indicated that in vitro capacitation induced significant changes in the distribution of motile sperm subpopulations, being affected by cryopreservation. Notwithstanding, the subpopulation which probably represents hyperactivated-like spermatozoa (SP4) also increased in frozen-thawed samples, occurring faster and simultaneously to the increment of mitochondrial activity and tyrosine phosphorylation of different sperm regions.Patricia Peris-FrauIrene Sánchez-AjofrínAlicia Martín MaestroCarolina MasideDaniela Alejandra Medina-ChávezOlga García-ÁlvarezMaría del Rocío Fernández-SantosVidal MontoroJosé Julián GardeManuel RamónAna Josefa SolerMDPI AGarticlecomputer-assisted sperm analysiscytometrysubpopulationssperm capacitationsperm cryopreservationtyrosine phosphorylationBiology (General)QH301-705.5ENBiology, Vol 10, Iss 1213, p 1213 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	computer-assisted sperm analysis cytometry subpopulations sperm capacitation sperm cryopreservation tyrosine phosphorylation Biology (General) QH301-705.5
spellingShingle	computer-assisted sperm analysis cytometry subpopulations sperm capacitation sperm cryopreservation tyrosine phosphorylation Biology (General) QH301-705.5 Patricia Peris-Frau Irene Sánchez-Ajofrín Alicia Martín Maestro Carolina Maside Daniela Alejandra Medina-Chávez Olga García-Álvarez María del Rocío Fernández-Santos Vidal Montoro José Julián Garde Manuel Ramón Ana Josefa Soler Impact of Cryopreservation on Motile Subpopulations and Tyrosine-Phosphorylated Regions of Ram Spermatozoa during Capacitating Conditions
description	The heterogeneous nature of ejaculates highlights the relevance of studying the behavior of different sperm subpopulations. Changes in sperm motility and the increase in tyrosine phosphorylation are key events that usually occur during capacitation and can be modified by the cryopreservation process. However, the relationship between both events remains poorly defined throughout capacitation in the different sperm subpopulations. Fresh and frozen-thawed spermatozoa were incubated in capacitating (CAP) and non-capacitating (NC) media up to 240 min. Sperm kinematics, tyrosine phosphorylation and mitochondrial activity were measured by the CASA system and imaging flow cytometry. Four motile sperm subpopulations (SP) were identified in fresh and frozen-thawed ram semen after the cluster analysis. Incubation under CAP conditions over time led to greater changes in the percentage of spermatozoa included in each subpopulation compared to NC conditions, being different between fresh and frozen-thawed spermatozoa. The SP1, characterized by slow spermatozoa, progressively increased after 15 min in frozen-thawed samples incubated in both media but not in fresh ones. The SP4, characterized by fast and non-linear spermatozoa, showed a marked increase during CAP, but not under NC conditions, occurring more rapidly in frozen-thawed spermatozoa. This subpopulation (SP4) was also the only one positively and strongly correlated with mitochondrial activity and all phosphorylated sperm regions during capacitation, either in fresh or frozen-thawed samples. Our results indicated that in vitro capacitation induced significant changes in the distribution of motile sperm subpopulations, being affected by cryopreservation. Notwithstanding, the subpopulation which probably represents hyperactivated-like spermatozoa (SP4) also increased in frozen-thawed samples, occurring faster and simultaneously to the increment of mitochondrial activity and tyrosine phosphorylation of different sperm regions.
format	article
author	Patricia Peris-Frau Irene Sánchez-Ajofrín Alicia Martín Maestro Carolina Maside Daniela Alejandra Medina-Chávez Olga García-Álvarez María del Rocío Fernández-Santos Vidal Montoro José Julián Garde Manuel Ramón Ana Josefa Soler
author_facet	Patricia Peris-Frau Irene Sánchez-Ajofrín Alicia Martín Maestro Carolina Maside Daniela Alejandra Medina-Chávez Olga García-Álvarez María del Rocío Fernández-Santos Vidal Montoro José Julián Garde Manuel Ramón Ana Josefa Soler
author_sort	Patricia Peris-Frau
title	Impact of Cryopreservation on Motile Subpopulations and Tyrosine-Phosphorylated Regions of Ram Spermatozoa during Capacitating Conditions
title_short	Impact of Cryopreservation on Motile Subpopulations and Tyrosine-Phosphorylated Regions of Ram Spermatozoa during Capacitating Conditions
title_full	Impact of Cryopreservation on Motile Subpopulations and Tyrosine-Phosphorylated Regions of Ram Spermatozoa during Capacitating Conditions
title_fullStr	Impact of Cryopreservation on Motile Subpopulations and Tyrosine-Phosphorylated Regions of Ram Spermatozoa during Capacitating Conditions
title_full_unstemmed	Impact of Cryopreservation on Motile Subpopulations and Tyrosine-Phosphorylated Regions of Ram Spermatozoa during Capacitating Conditions
title_sort	impact of cryopreservation on motile subpopulations and tyrosine-phosphorylated regions of ram spermatozoa during capacitating conditions
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/f6abda8580574ed7a7a8ca969d7a03a3
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Impact of Cryopreservation on Motile Subpopulations and Tyrosine-Phosphorylated Regions of Ram Spermatozoa during Capacitating Conditions

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