Evidence that human epididymal protein ARP plays a role in gamete fusion through complementary sites on the surface of the human egg
Human epididymal sperm protein ARP, a member of the cysteine-rich secretory proteins (CRISP) family, exhibits significant homology with rat epididymal protein DE, a candidate molecule for mediating sperm-egg fusion in rodents. The aim of this study was to investigate the involvement of ARP in human...
Guardado en:
| Autores principales: | , , , , , , , , |
|---|---|
| Formato: | JOUR |
| Materias: | |
| Acceso en línea: | http://hdl.handle.net/20.500.12110/paper_00063363_v65_n4_p1000_Cohen |
| Aporte de: |
| id |
todo:paper_00063363_v65_n4_p1000_Cohen |
|---|---|
| record_format |
dspace |
| spelling |
todo:paper_00063363_v65_n4_p1000_Cohen2023-10-03T14:04:51Z Evidence that human epididymal protein ARP plays a role in gamete fusion through complementary sites on the surface of the human egg Cohen, D.J. Ellerman, D.A. Busso, D. Morgenfeld, M.M. Piazza, A.D. Hayashi, M. Young, E.T. Kasahara, M. Cuasnicu, P.S. Epididymis Fertilization Gamete biology Ovum Sperm calcium ionophore human epididymal sperm protein ARP polyclonal antibody rat epididymal sperm protein DE recombinant protein secretory protein unclassified drug acrosome reaction animal cell article cell fusion cell viability concentration response controlled study extraction female fertilization hamster human human cell immunofluorescence male metabolic inhibition nonhuman normal human oocyte priority journal sequence homology sperm spermatozoon capacitation spermatozoon motility spermatozoon penetration surface property zona pellucida Human epididymal sperm protein ARP, a member of the cysteine-rich secretory proteins (CRISP) family, exhibits significant homology with rat epididymal protein DE, a candidate molecule for mediating sperm-egg fusion in rodents. The aim of this study was to investigate the involvement of ARP in human gamete fusion. Sequential extraction of proteins from ejaculated human sperm revealed the existence of a population of ARP that is tightly associated with the sperm surface and thus, potentially capable of participating in gamete interaction. Exposure of capacitated human sperm to a polyclonal antibody against recombinant ARP (anti-ARP) produced a significant and concentration-dependent inhibition in the ability of human sperm to penetrate zona-free hamster eggs. This inhibition was not due to a deleterious effect on the gametes because anti-ARP affected neither sperm viability or motility, nor egg penetrability. The antibody did not inhibit the occurrence of spontaneous or Ca 2+ ionophore-induced acrosome reaction, nor did it inhibit the ability of sperm to bind to the oolema, supporting a specific inhibition of the antibody at the sperm-egg fusion level. As a relevant evidence for a role of ARP in gamete fusion, the existence of complementary sites for this protein on the surface of human eggs was investigated. Experiments in which zona-free human oocytes discarded from in vitro fertilization programs were exposed to ARP, fixed, and subjected to indirect immunofluorescence revealed the presence of specific ARP-binding sites on the entire surface of the human egg, in agreement with the fusogenic properties of the human oolema. Together, these results strongly support the participation of ARP in the sperm-egg fusion process, suggesting that this protein would be the functional homologue of DE in humans. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_00063363_v65_n4_p1000_Cohen |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Epididymis Fertilization Gamete biology Ovum Sperm calcium ionophore human epididymal sperm protein ARP polyclonal antibody rat epididymal sperm protein DE recombinant protein secretory protein unclassified drug acrosome reaction animal cell article cell fusion cell viability concentration response controlled study extraction female fertilization hamster human human cell immunofluorescence male metabolic inhibition nonhuman normal human oocyte priority journal sequence homology sperm spermatozoon capacitation spermatozoon motility spermatozoon penetration surface property zona pellucida |
| spellingShingle |
Epididymis Fertilization Gamete biology Ovum Sperm calcium ionophore human epididymal sperm protein ARP polyclonal antibody rat epididymal sperm protein DE recombinant protein secretory protein unclassified drug acrosome reaction animal cell article cell fusion cell viability concentration response controlled study extraction female fertilization hamster human human cell immunofluorescence male metabolic inhibition nonhuman normal human oocyte priority journal sequence homology sperm spermatozoon capacitation spermatozoon motility spermatozoon penetration surface property zona pellucida Cohen, D.J. Ellerman, D.A. Busso, D. Morgenfeld, M.M. Piazza, A.D. Hayashi, M. Young, E.T. Kasahara, M. Cuasnicu, P.S. Evidence that human epididymal protein ARP plays a role in gamete fusion through complementary sites on the surface of the human egg |
| topic_facet |
Epididymis Fertilization Gamete biology Ovum Sperm calcium ionophore human epididymal sperm protein ARP polyclonal antibody rat epididymal sperm protein DE recombinant protein secretory protein unclassified drug acrosome reaction animal cell article cell fusion cell viability concentration response controlled study extraction female fertilization hamster human human cell immunofluorescence male metabolic inhibition nonhuman normal human oocyte priority journal sequence homology sperm spermatozoon capacitation spermatozoon motility spermatozoon penetration surface property zona pellucida |
| description |
Human epididymal sperm protein ARP, a member of the cysteine-rich secretory proteins (CRISP) family, exhibits significant homology with rat epididymal protein DE, a candidate molecule for mediating sperm-egg fusion in rodents. The aim of this study was to investigate the involvement of ARP in human gamete fusion. Sequential extraction of proteins from ejaculated human sperm revealed the existence of a population of ARP that is tightly associated with the sperm surface and thus, potentially capable of participating in gamete interaction. Exposure of capacitated human sperm to a polyclonal antibody against recombinant ARP (anti-ARP) produced a significant and concentration-dependent inhibition in the ability of human sperm to penetrate zona-free hamster eggs. This inhibition was not due to a deleterious effect on the gametes because anti-ARP affected neither sperm viability or motility, nor egg penetrability. The antibody did not inhibit the occurrence of spontaneous or Ca 2+ ionophore-induced acrosome reaction, nor did it inhibit the ability of sperm to bind to the oolema, supporting a specific inhibition of the antibody at the sperm-egg fusion level. As a relevant evidence for a role of ARP in gamete fusion, the existence of complementary sites for this protein on the surface of human eggs was investigated. Experiments in which zona-free human oocytes discarded from in vitro fertilization programs were exposed to ARP, fixed, and subjected to indirect immunofluorescence revealed the presence of specific ARP-binding sites on the entire surface of the human egg, in agreement with the fusogenic properties of the human oolema. Together, these results strongly support the participation of ARP in the sperm-egg fusion process, suggesting that this protein would be the functional homologue of DE in humans. |
| format |
JOUR |
| author |
Cohen, D.J. Ellerman, D.A. Busso, D. Morgenfeld, M.M. Piazza, A.D. Hayashi, M. Young, E.T. Kasahara, M. Cuasnicu, P.S. |
| author_facet |
Cohen, D.J. Ellerman, D.A. Busso, D. Morgenfeld, M.M. Piazza, A.D. Hayashi, M. Young, E.T. Kasahara, M. Cuasnicu, P.S. |
| author_sort |
Cohen, D.J. |
| title |
Evidence that human epididymal protein ARP plays a role in gamete fusion through complementary sites on the surface of the human egg |
| title_short |
Evidence that human epididymal protein ARP plays a role in gamete fusion through complementary sites on the surface of the human egg |
| title_full |
Evidence that human epididymal protein ARP plays a role in gamete fusion through complementary sites on the surface of the human egg |
| title_fullStr |
Evidence that human epididymal protein ARP plays a role in gamete fusion through complementary sites on the surface of the human egg |
| title_full_unstemmed |
Evidence that human epididymal protein ARP plays a role in gamete fusion through complementary sites on the surface of the human egg |
| title_sort |
evidence that human epididymal protein arp plays a role in gamete fusion through complementary sites on the surface of the human egg |
| url |
http://hdl.handle.net/20.500.12110/paper_00063363_v65_n4_p1000_Cohen |
| work_keys_str_mv |
AT cohendj evidencethathumanepididymalproteinarpplaysaroleingametefusionthroughcomplementarysitesonthesurfaceofthehumanegg AT ellermanda evidencethathumanepididymalproteinarpplaysaroleingametefusionthroughcomplementarysitesonthesurfaceofthehumanegg AT bussod evidencethathumanepididymalproteinarpplaysaroleingametefusionthroughcomplementarysitesonthesurfaceofthehumanegg AT morgenfeldmm evidencethathumanepididymalproteinarpplaysaroleingametefusionthroughcomplementarysitesonthesurfaceofthehumanegg AT piazzaad evidencethathumanepididymalproteinarpplaysaroleingametefusionthroughcomplementarysitesonthesurfaceofthehumanegg AT hayashim evidencethathumanepididymalproteinarpplaysaroleingametefusionthroughcomplementarysitesonthesurfaceofthehumanegg AT younget evidencethathumanepididymalproteinarpplaysaroleingametefusionthroughcomplementarysitesonthesurfaceofthehumanegg AT kasaharam evidencethathumanepididymalproteinarpplaysaroleingametefusionthroughcomplementarysitesonthesurfaceofthehumanegg AT cuasnicups evidencethathumanepididymalproteinarpplaysaroleingametefusionthroughcomplementarysitesonthesurfaceofthehumanegg |
| _version_ |
1807315381233123328 |