Bicarbonate-Stimulated Membrane Reorganization in Stallion Spermatozoa

Classical in vitro fertilization (IVF) is still poorly successful in horses. This lack of success is thought to be due primarily to inadequate capacitation of stallion spermatozoa under in vitro conditions. In species in which IVF is successful, bicarbonate, calcium, and albumin are considered the k...

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Autores principales:	Paula Piccolo Maitan, Elizabeth G. Bromfield, Romy Hoogendijk, Miguel Ricardo Leung, Tzviya Zeev-Ben-Mordehai, Chris H. van de Lest, Jeroen W. A. Jansen, Bart Leemans, José Domingos Guimarães, Tom A. E. Stout, Bart M. Gadella, Heiko Henning
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	spermatozoa capacitation membrane lipid bicarbonate (HCO3−) fertilization Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/d3b8d0ea79974a7089f3a5f2dc86588e
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spelling	oai:doaj.org-article:d3b8d0ea79974a7089f3a5f2dc86588e2021-11-17T05:58:51ZBicarbonate-Stimulated Membrane Reorganization in Stallion Spermatozoa2296-634X10.3389/fcell.2021.772254https://doaj.org/article/d3b8d0ea79974a7089f3a5f2dc86588e2021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fcell.2021.772254/fullhttps://doaj.org/toc/2296-634XClassical in vitro fertilization (IVF) is still poorly successful in horses. This lack of success is thought to be due primarily to inadequate capacitation of stallion spermatozoa under in vitro conditions. In species in which IVF is successful, bicarbonate, calcium, and albumin are considered the key components that enable a gradual reorganization of the sperm plasma membrane that allows the spermatozoa to undergo an acrosome reaction and fertilize the oocyte. The aim of this work was to comprehensively examine contributors to stallion sperm capacitation by investigating bicarbonate-induced membrane remodelling steps, and elucidating the contribution of cAMP signalling to these events. In the presence of capacitating media containing bicarbonate, a significant increase in plasma membrane fluidity was readily detected using merocyanine 540 staining in the majority of viable spermatozoa within 15 min of bicarbonate exposure. Specific inhibition of soluble adenylyl cyclase (sAC) in the presence of bicarbonate by LRE1 significantly reduced the number of viable sperm with high membrane fluidity. This suggests a vital role for sAC-mediated cAMP production in the regulation of membrane fluidity. Cryo-electron tomography of viable cells with high membrane fluidity revealed a range of membrane remodelling intermediates, including destabilized membranes and zones with close apposition of the plasma membrane and the outer acrosomal membrane. However, lipidomic analysis of equivalent viable spermatozoa with high membrane fluidity demonstrated that this phenomenon was neither accompanied by a gross change in the phospholipid composition of stallion sperm membranes nor detectable sterol efflux (p > 0.05). After an early increase in membrane fluidity, a significant and cAMP-dependent increase in viable sperm with phosphatidylserine (PS), but not phosphatidylethanolamine (PE) exposure was noted. While the events observed partly resemble findings from the in vitro capacitation of sperm from other mammalian species, the lack of cholesterol removal appears to be an equine-specific phenomenon. This research will assist in the development of a defined medium for the capacitation of stallion sperm and will facilitate progress toward a functional IVF protocol for horse gametes.Paula Piccolo MaitanPaula Piccolo MaitanElizabeth G. BromfieldElizabeth G. BromfieldRomy HoogendijkMiguel Ricardo LeungTzviya Zeev-Ben-MordehaiChris H. van de LestJeroen W. A. JansenBart LeemansJosé Domingos GuimarãesTom A. E. StoutBart M. GadellaBart M. GadellaHeiko HenningFrontiers Media S.A.articlespermatozoacapacitationmembranelipidbicarbonate (HCO3−)fertilizationBiology (General)QH301-705.5ENFrontiers in Cell and Developmental Biology, Vol 9 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	spermatozoa capacitation membrane lipid bicarbonate (HCO3−) fertilization Biology (General) QH301-705.5
spellingShingle	spermatozoa capacitation membrane lipid bicarbonate (HCO3−) fertilization Biology (General) QH301-705.5 Paula Piccolo Maitan Paula Piccolo Maitan Elizabeth G. Bromfield Elizabeth G. Bromfield Romy Hoogendijk Miguel Ricardo Leung Tzviya Zeev-Ben-Mordehai Chris H. van de Lest Jeroen W. A. Jansen Bart Leemans José Domingos Guimarães Tom A. E. Stout Bart M. Gadella Bart M. Gadella Heiko Henning Bicarbonate-Stimulated Membrane Reorganization in Stallion Spermatozoa
description	Classical in vitro fertilization (IVF) is still poorly successful in horses. This lack of success is thought to be due primarily to inadequate capacitation of stallion spermatozoa under in vitro conditions. In species in which IVF is successful, bicarbonate, calcium, and albumin are considered the key components that enable a gradual reorganization of the sperm plasma membrane that allows the spermatozoa to undergo an acrosome reaction and fertilize the oocyte. The aim of this work was to comprehensively examine contributors to stallion sperm capacitation by investigating bicarbonate-induced membrane remodelling steps, and elucidating the contribution of cAMP signalling to these events. In the presence of capacitating media containing bicarbonate, a significant increase in plasma membrane fluidity was readily detected using merocyanine 540 staining in the majority of viable spermatozoa within 15 min of bicarbonate exposure. Specific inhibition of soluble adenylyl cyclase (sAC) in the presence of bicarbonate by LRE1 significantly reduced the number of viable sperm with high membrane fluidity. This suggests a vital role for sAC-mediated cAMP production in the regulation of membrane fluidity. Cryo-electron tomography of viable cells with high membrane fluidity revealed a range of membrane remodelling intermediates, including destabilized membranes and zones with close apposition of the plasma membrane and the outer acrosomal membrane. However, lipidomic analysis of equivalent viable spermatozoa with high membrane fluidity demonstrated that this phenomenon was neither accompanied by a gross change in the phospholipid composition of stallion sperm membranes nor detectable sterol efflux (p > 0.05). After an early increase in membrane fluidity, a significant and cAMP-dependent increase in viable sperm with phosphatidylserine (PS), but not phosphatidylethanolamine (PE) exposure was noted. While the events observed partly resemble findings from the in vitro capacitation of sperm from other mammalian species, the lack of cholesterol removal appears to be an equine-specific phenomenon. This research will assist in the development of a defined medium for the capacitation of stallion sperm and will facilitate progress toward a functional IVF protocol for horse gametes.
format	article
author	Paula Piccolo Maitan Paula Piccolo Maitan Elizabeth G. Bromfield Elizabeth G. Bromfield Romy Hoogendijk Miguel Ricardo Leung Tzviya Zeev-Ben-Mordehai Chris H. van de Lest Jeroen W. A. Jansen Bart Leemans José Domingos Guimarães Tom A. E. Stout Bart M. Gadella Bart M. Gadella Heiko Henning
author_facet	Paula Piccolo Maitan Paula Piccolo Maitan Elizabeth G. Bromfield Elizabeth G. Bromfield Romy Hoogendijk Miguel Ricardo Leung Tzviya Zeev-Ben-Mordehai Chris H. van de Lest Jeroen W. A. Jansen Bart Leemans José Domingos Guimarães Tom A. E. Stout Bart M. Gadella Bart M. Gadella Heiko Henning
author_sort	Paula Piccolo Maitan
title	Bicarbonate-Stimulated Membrane Reorganization in Stallion Spermatozoa
title_short	Bicarbonate-Stimulated Membrane Reorganization in Stallion Spermatozoa
title_full	Bicarbonate-Stimulated Membrane Reorganization in Stallion Spermatozoa
title_fullStr	Bicarbonate-Stimulated Membrane Reorganization in Stallion Spermatozoa
title_full_unstemmed	Bicarbonate-Stimulated Membrane Reorganization in Stallion Spermatozoa
title_sort	bicarbonate-stimulated membrane reorganization in stallion spermatozoa
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/d3b8d0ea79974a7089f3a5f2dc86588e
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Bicarbonate-Stimulated Membrane Reorganization in Stallion Spermatozoa

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