Human fertilization: Epididymal hCRISP1 mediates sperm-zona pellucida binding through its interaction with ZP3

Human epididymal CRISP1 (hCRISP1) associates with sperm during maturation and participates in gamete fusion through egg complementary sites. Its homology with both rodent epididymal CRISP1 and CRISP4 reported to participate in the previous stage of sperm binding to the zona pellucida (ZP), led us to...

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Autores principales:	Maldera, J.A., Muñoz, M.W., Chirinos, M., Busso, D., Raffo, F.G.E., Battistone, M.A., Blaquier, J.A., Larrea, F., Cuasnicu, P.S.
Formato:	JOUR
Materias:	Fertilization HCRISP1 Oocyte Sperm Zona pellucida CRISP1 protein polyclonal antibody protein recombinant protein unclassified drug zona pellucida protein 2 zona pellucida protein 3 zona pellucida protein 4 acrosome reaction animal cell article artificial insemination binding site controlled study enzyme linked immunosorbent assay female fertilization human human cell immunofluorescence insect cell male nonhuman priority journal protein expression protein phosphorylation protein protein interaction rat semen analysis sperm spermatozoon capacitation spermatozoon motility zona pellucida hCRISP1 oocyte Acrosome Reaction Adult Antibodies Binding Sites Binding, Competitive Egg Proteins Epididymis Female Gene Expression Regulation Humans Male Membrane Glycoproteins Protein Binding Receptors, Cell Surface Recombinant Proteins Signal Transduction Sperm Capacitation Spermatozoa Zona Pellucida
Acceso en línea:	http://hdl.handle.net/20.500.12110/paper_13609947_v20_n4_p341_Maldera
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	todo:paper_13609947_v20_n4_p341_Maldera
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spelling	todo:paper_13609947_v20_n4_p341_Maldera2023-10-03T16:10:50Z Human fertilization: Epididymal hCRISP1 mediates sperm-zona pellucida binding through its interaction with ZP3 Maldera, J.A. Muñoz, M.W. Chirinos, M. Busso, D. Raffo, F.G.E. Battistone, M.A. Blaquier, J.A. Larrea, F. Cuasnicu, P.S. Fertilization HCRISP1 Oocyte Sperm Zona pellucida CRISP1 protein polyclonal antibody protein recombinant protein unclassified drug zona pellucida protein 2 zona pellucida protein 3 zona pellucida protein 4 acrosome reaction animal cell article artificial insemination binding site controlled study enzyme linked immunosorbent assay female fertilization human human cell immunofluorescence insect cell male nonhuman priority journal protein expression protein phosphorylation protein protein interaction rat semen analysis sperm spermatozoon capacitation spermatozoon motility zona pellucida fertilization hCRISP1 oocyte sperm zona pellucida Acrosome Reaction Adult Antibodies Binding Sites Binding, Competitive Egg Proteins Epididymis Female Gene Expression Regulation Humans Male Membrane Glycoproteins Protein Binding Receptors, Cell Surface Recombinant Proteins Signal Transduction Sperm Capacitation Spermatozoa Zona Pellucida Human epididymal CRISP1 (hCRISP1) associates with sperm during maturation and participates in gamete fusion through egg complementary sites. Its homology with both rodent epididymal CRISP1 and CRISP4 reported to participate in the previous stage of sperm binding to the zona pellucida (ZP), led us to further investigate the functional role of hCRISP1 by studying its involvement in human sperm- ZP interaction. Human hemizona (HZ) were inseminated with human capacitated sperm in the presence of either anti-hCRISP1 polyclonal antibody to inhibit sperm hCRISP1, or bacterially-expressed hCRISP1 (rec-hCRISP1) to block putative hCRISP1 binding sites in the ZP. Results revealed that both anti-hCRISP1 and rec-hCRISP1 produced a significant inhibition in the number of sperm bound per HZ compared with the corresponding controls. The finding that neither anti-hCRISP1 nor rec-hCRISP1 affected capacitation-associated events (i.e. sperm motility, protein tyrosine phosphorylation or acrosome reaction) supports a specific inhibition at the sperm-egg interaction level. Moreover, immunofluorescence experiments using human ZP-intact eggs revealed the presence of complementary sites for hCRISP1 in the ZP. To identify the ligand of hCRISP1 in the ZP, human recombinant proteins ZP2, ZP3 and ZP4 expressed in insect cells were co-incubated with hCRISP1 and protein- protein interaction was analyzed by ELISA. Results revealed that rec-hCRISP1 mainly interacted with ZP3 in a dose-dependent and saturable manner, supporting the specificity of this interaction. Altogether, these results indicate that hCRISP1 is a multifunctional protein involved not only in sperm-egg fusion but also in the previous stage of sperm-ZP binding through its specific interaction with human ZP3. © The Author 2013. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_13609947_v20_n4_p341_Maldera
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	Fertilization HCRISP1 Oocyte Sperm Zona pellucida CRISP1 protein polyclonal antibody protein recombinant protein unclassified drug zona pellucida protein 2 zona pellucida protein 3 zona pellucida protein 4 acrosome reaction animal cell article artificial insemination binding site controlled study enzyme linked immunosorbent assay female fertilization human human cell immunofluorescence insect cell male nonhuman priority journal protein expression protein phosphorylation protein protein interaction rat semen analysis sperm spermatozoon capacitation spermatozoon motility zona pellucida fertilization hCRISP1 oocyte sperm zona pellucida Acrosome Reaction Adult Antibodies Binding Sites Binding, Competitive Egg Proteins Epididymis Female Gene Expression Regulation Humans Male Membrane Glycoproteins Protein Binding Receptors, Cell Surface Recombinant Proteins Signal Transduction Sperm Capacitation Spermatozoa Zona Pellucida
spellingShingle	Fertilization HCRISP1 Oocyte Sperm Zona pellucida CRISP1 protein polyclonal antibody protein recombinant protein unclassified drug zona pellucida protein 2 zona pellucida protein 3 zona pellucida protein 4 acrosome reaction animal cell article artificial insemination binding site controlled study enzyme linked immunosorbent assay female fertilization human human cell immunofluorescence insect cell male nonhuman priority journal protein expression protein phosphorylation protein protein interaction rat semen analysis sperm spermatozoon capacitation spermatozoon motility zona pellucida fertilization hCRISP1 oocyte sperm zona pellucida Acrosome Reaction Adult Antibodies Binding Sites Binding, Competitive Egg Proteins Epididymis Female Gene Expression Regulation Humans Male Membrane Glycoproteins Protein Binding Receptors, Cell Surface Recombinant Proteins Signal Transduction Sperm Capacitation Spermatozoa Zona Pellucida Maldera, J.A. Muñoz, M.W. Chirinos, M. Busso, D. Raffo, F.G.E. Battistone, M.A. Blaquier, J.A. Larrea, F. Cuasnicu, P.S. Human fertilization: Epididymal hCRISP1 mediates sperm-zona pellucida binding through its interaction with ZP3
topic_facet	Fertilization HCRISP1 Oocyte Sperm Zona pellucida CRISP1 protein polyclonal antibody protein recombinant protein unclassified drug zona pellucida protein 2 zona pellucida protein 3 zona pellucida protein 4 acrosome reaction animal cell article artificial insemination binding site controlled study enzyme linked immunosorbent assay female fertilization human human cell immunofluorescence insect cell male nonhuman priority journal protein expression protein phosphorylation protein protein interaction rat semen analysis sperm spermatozoon capacitation spermatozoon motility zona pellucida fertilization hCRISP1 oocyte sperm zona pellucida Acrosome Reaction Adult Antibodies Binding Sites Binding, Competitive Egg Proteins Epididymis Female Gene Expression Regulation Humans Male Membrane Glycoproteins Protein Binding Receptors, Cell Surface Recombinant Proteins Signal Transduction Sperm Capacitation Spermatozoa Zona Pellucida
description	Human epididymal CRISP1 (hCRISP1) associates with sperm during maturation and participates in gamete fusion through egg complementary sites. Its homology with both rodent epididymal CRISP1 and CRISP4 reported to participate in the previous stage of sperm binding to the zona pellucida (ZP), led us to further investigate the functional role of hCRISP1 by studying its involvement in human sperm- ZP interaction. Human hemizona (HZ) were inseminated with human capacitated sperm in the presence of either anti-hCRISP1 polyclonal antibody to inhibit sperm hCRISP1, or bacterially-expressed hCRISP1 (rec-hCRISP1) to block putative hCRISP1 binding sites in the ZP. Results revealed that both anti-hCRISP1 and rec-hCRISP1 produced a significant inhibition in the number of sperm bound per HZ compared with the corresponding controls. The finding that neither anti-hCRISP1 nor rec-hCRISP1 affected capacitation-associated events (i.e. sperm motility, protein tyrosine phosphorylation or acrosome reaction) supports a specific inhibition at the sperm-egg interaction level. Moreover, immunofluorescence experiments using human ZP-intact eggs revealed the presence of complementary sites for hCRISP1 in the ZP. To identify the ligand of hCRISP1 in the ZP, human recombinant proteins ZP2, ZP3 and ZP4 expressed in insect cells were co-incubated with hCRISP1 and protein- protein interaction was analyzed by ELISA. Results revealed that rec-hCRISP1 mainly interacted with ZP3 in a dose-dependent and saturable manner, supporting the specificity of this interaction. Altogether, these results indicate that hCRISP1 is a multifunctional protein involved not only in sperm-egg fusion but also in the previous stage of sperm-ZP binding through its specific interaction with human ZP3. © The Author 2013. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved.
format	JOUR
author	Maldera, J.A. Muñoz, M.W. Chirinos, M. Busso, D. Raffo, F.G.E. Battistone, M.A. Blaquier, J.A. Larrea, F. Cuasnicu, P.S.
author_facet	Maldera, J.A. Muñoz, M.W. Chirinos, M. Busso, D. Raffo, F.G.E. Battistone, M.A. Blaquier, J.A. Larrea, F. Cuasnicu, P.S.
author_sort	Maldera, J.A.
title	Human fertilization: Epididymal hCRISP1 mediates sperm-zona pellucida binding through its interaction with ZP3
title_short	Human fertilization: Epididymal hCRISP1 mediates sperm-zona pellucida binding through its interaction with ZP3
title_full	Human fertilization: Epididymal hCRISP1 mediates sperm-zona pellucida binding through its interaction with ZP3
title_fullStr	Human fertilization: Epididymal hCRISP1 mediates sperm-zona pellucida binding through its interaction with ZP3
title_full_unstemmed	Human fertilization: Epididymal hCRISP1 mediates sperm-zona pellucida binding through its interaction with ZP3
title_sort	human fertilization: epididymal hcrisp1 mediates sperm-zona pellucida binding through its interaction with zp3
url	http://hdl.handle.net/20.500.12110/paper_13609947_v20_n4_p341_Maldera
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Human fertilization: Epididymal hCRISP1 mediates sperm-zona pellucida binding through its interaction with ZP3

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