Further evidence on the role of heparan sulfate as protamine acceptor during the decondensation of human spermatozoa

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Background: Human spermatozoa decondense in vitro upon exposure to heparin and glutathione. Glutathione is also the disulfide bond reducer in vivo, and heparan sulfate, a functional analogue of heparin, has been proposed as the protamine acceptor. The aim of this study was to evaluate the decondensi...

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Autores principales: Romanato, M., Regueira, E., Cameo, M.S., Baldini, C., Calvo, L., Calvo, J.C.
Formato: Artículo publishedVersion
Lenguaje:Inglés
Publicado: 2005
Materias:
Heparan sulfate
Protamine
Sperm nuclear decondensation
glutaraldehyde
glutathione
glycosaminoglycan
heparan sulfate
heparin
protamine
article
carbohydrate analysis
cell isolation
chromatin condensation
drug activity
human
human cell
immunocytochemistry
in vitro study
male
phase contrast microscopy
polyacrylamide gel electrophoresis
protein analysis
protein isolation
protein secretion
semen analysis
spermatozoon capacitation
spermatozoon density
Acceso en línea:http://hdl.handle.net/20.500.12110/paper_02681161_v20_n10_p2784_Romanato
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Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires
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spelling paperaa:paper_02681161_v20_n10_p2784_Romanato2023-06-12T16:47:11Z Further evidence on the role of heparan sulfate as protamine acceptor during the decondensation of human spermatozoa Hum. Reprod. 2005;20(10):2784-2789 Romanato, M. Regueira, E. Cameo, M.S. Baldini, C. Calvo, L. Calvo, J.C. Heparan sulfate Protamine Sperm nuclear decondensation glutaraldehyde glutathione glycosaminoglycan heparan sulfate heparin protamine article carbohydrate analysis cell isolation chromatin condensation drug activity human human cell immunocytochemistry in vitro study male phase contrast microscopy polyacrylamide gel electrophoresis protein analysis protein isolation protein secretion semen analysis spermatozoon capacitation spermatozoon density Background: Human spermatozoa decondense in vitro upon exposure to heparin and glutathione. Glutathione is also the disulfide bond reducer in vivo, and heparan sulfate, a functional analogue of heparin, has been proposed as the protamine acceptor. The aim of this study was to evaluate the decondensing ability of chemically modified heparins and different glycosaminoglycans (GAGs) on isolated sperm nuclei in vitro, and to analyse the possible role of different GAGs as protamine acceptors. Methods: Capacitated spermatozoa and isolated sperm nuclei from normospermic semen samples were decondensed in the presence of heparin (or its equivalent) and glutathione. After fixation with glutaraldehyde, the percentage of decondensed spermatozoa and nuclei was determined under phase-contrast. Proteins were extracted from sperm nuclei previously incubated in the presence of gluhathione and different GAGs by incubation with urea-β-meracptoethanol-NaCl, and analysed by acid polyacrylamide gel electrophoresis. Results: The ability of desulfated heparins and other GAGs to decondense isolated nuclei mirrored exactly the decondensation of capacitated spermatozoa, the only difference being the level of maximum decondensation achieved. Heparan sulfate and heparin, but not other GAGs, were able to release protamines from sperm chromatin. Conclusions: Heparan sulfate could be functioning as protamine acceptor in vivo during human sperm nuclear decondensation. © The Author 2005. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. Fil:Romanato, M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Regueira, E. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Calvo, J.C. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2005 info:eu-repo/semantics/article info:ar-repo/semantics/artículo info:eu-repo/semantics/publishedVersion application/pdf eng info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_02681161_v20_n10_p2784_Romanato
institution Universidad de Buenos Aires
institution_str I-28
repository_str R-134
collection Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
language Inglés
orig_language_str_mv eng
topic Heparan sulfate
Protamine
Sperm nuclear decondensation
glutaraldehyde
glutathione
glycosaminoglycan
heparan sulfate
heparin
protamine
article
carbohydrate analysis
cell isolation
chromatin condensation
drug activity
human
human cell
immunocytochemistry
in vitro study
male
phase contrast microscopy
polyacrylamide gel electrophoresis
protein analysis
protein isolation
protein secretion
semen analysis
spermatozoon capacitation
spermatozoon density
spellingShingle Heparan sulfate
Protamine
Sperm nuclear decondensation
glutaraldehyde
glutathione
glycosaminoglycan
heparan sulfate
heparin
protamine
article
carbohydrate analysis
cell isolation
chromatin condensation
drug activity
human
human cell
immunocytochemistry
in vitro study
male
phase contrast microscopy
polyacrylamide gel electrophoresis
protein analysis
protein isolation
protein secretion
semen analysis
spermatozoon capacitation
spermatozoon density
Romanato, M.
Regueira, E.
Cameo, M.S.
Baldini, C.
Calvo, L.
Calvo, J.C.
Further evidence on the role of heparan sulfate as protamine acceptor during the decondensation of human spermatozoa
topic_facet Heparan sulfate
Protamine
Sperm nuclear decondensation
glutaraldehyde
glutathione
glycosaminoglycan
heparan sulfate
heparin
protamine
article
carbohydrate analysis
cell isolation
chromatin condensation
drug activity
human
human cell
immunocytochemistry
in vitro study
male
phase contrast microscopy
polyacrylamide gel electrophoresis
protein analysis
protein isolation
protein secretion
semen analysis
spermatozoon capacitation
spermatozoon density
description Background: Human spermatozoa decondense in vitro upon exposure to heparin and glutathione. Glutathione is also the disulfide bond reducer in vivo, and heparan sulfate, a functional analogue of heparin, has been proposed as the protamine acceptor. The aim of this study was to evaluate the decondensing ability of chemically modified heparins and different glycosaminoglycans (GAGs) on isolated sperm nuclei in vitro, and to analyse the possible role of different GAGs as protamine acceptors. Methods: Capacitated spermatozoa and isolated sperm nuclei from normospermic semen samples were decondensed in the presence of heparin (or its equivalent) and glutathione. After fixation with glutaraldehyde, the percentage of decondensed spermatozoa and nuclei was determined under phase-contrast. Proteins were extracted from sperm nuclei previously incubated in the presence of gluhathione and different GAGs by incubation with urea-β-meracptoethanol-NaCl, and analysed by acid polyacrylamide gel electrophoresis. Results: The ability of desulfated heparins and other GAGs to decondense isolated nuclei mirrored exactly the decondensation of capacitated spermatozoa, the only difference being the level of maximum decondensation achieved. Heparan sulfate and heparin, but not other GAGs, were able to release protamines from sperm chromatin. Conclusions: Heparan sulfate could be functioning as protamine acceptor in vivo during human sperm nuclear decondensation. © The Author 2005. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved.
format Artículo
Artículo
publishedVersion
author Romanato, M.
Regueira, E.
Cameo, M.S.
Baldini, C.
Calvo, L.
Calvo, J.C.
author_facet Romanato, M.
Regueira, E.
Cameo, M.S.
Baldini, C.
Calvo, L.
Calvo, J.C.
author_sort Romanato, M.
title Further evidence on the role of heparan sulfate as protamine acceptor during the decondensation of human spermatozoa
title_short Further evidence on the role of heparan sulfate as protamine acceptor during the decondensation of human spermatozoa
title_full Further evidence on the role of heparan sulfate as protamine acceptor during the decondensation of human spermatozoa
title_fullStr Further evidence on the role of heparan sulfate as protamine acceptor during the decondensation of human spermatozoa
title_full_unstemmed Further evidence on the role of heparan sulfate as protamine acceptor during the decondensation of human spermatozoa
title_sort further evidence on the role of heparan sulfate as protamine acceptor during the decondensation of human spermatozoa
publishDate 2005
url http://hdl.handle.net/20.500.12110/paper_02681161_v20_n10_p2784_Romanato
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