Evidence for the involvement of testicular protein CRISP2 in mouse sperm-egg fusion
CRISP2, originally known as Tpx-1, is a cysteine-rich secretory protein specifically expressed in male haploid germ cells. Although likely to be involved in gamete interaction, evidence for a functional role of CRISP2 in fertilization still remains poor. In the present study, we used a mouse model t...
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2007
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| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00063363_v76_n4_p701_Busso http://hdl.handle.net/20.500.12110/paper_00063363_v76_n4_p701_Busso |
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paper:paper_00063363_v76_n4_p701_Busso2023-06-08T14:31:07Z Evidence for the involvement of testicular protein CRISP2 in mouse sperm-egg fusion Fertilization Ovum Sperm Testis cell protein isoprotein protein antibody protein CRISP1 protein CRISP2 recombinant protein secretory protein unclassified drug acrosome reaction animal cell animal tissue article cell fusion cell surface cellular distribution controlled study epididymis female fertilization fertilization in vitro human immunofluorescence male mouse nonhuman priority journal protein analysis protein binding protein function protein localization spermatozoon capacitation spermatozoon motility spermatozoon penetration testis vitelline membrane zona pellucida Animals Female Glycoproteins Male Mice Mice, Inbred Strains Sperm-Ovum Interactions Spermatozoa Testis Rattus CRISP2, originally known as Tpx-1, is a cysteine-rich secretory protein specifically expressed in male haploid germ cells. Although likely to be involved in gamete interaction, evidence for a functional role of CRISP2 in fertilization still remains poor. In the present study, we used a mouse model to examine the subcellular localization of CRISP2 in sperm and its involvement in the different stages of fertilization. Results from indirect immunofluorescence and protein extraction experiments indi-cated that mouse CRISP2 is an intraacrosomal component that remains associated with sperm after capacitation and the acrosome reaction (AR). In vitro fertilization assays using zona pellucida-intact mouse eggs showed that an antibody against the protein significantly decreased the percentage of penetrated eggs, with a coincident accumulation of perivitelline sperm. The failure to inhibit zona pellucida penetration excludes a detrimental effect of the antibody on sperm motility or the AR, supporting a specific participation of CRSSP2 at the sperm-egg fusion step. In agreement with this evidence, recombinant mouse CRISP2 (recCRISP2) specifically bound to the fusogenic area of mouse eggs, as previously reported for rat CRISP1, an epididymal protein involved in gamete fusion. In vitro competition investigations showed that incubation of mouse zona-free eggs with a fixed concentration of recCRISP2 and increasing amounts of rat CRISP1 reduced the binding of recCRISP2 to the egg, suggesting that the proteins interact with common complementary sites on the egg surface. Our findings indicate that testicular CRISP2, as observed for epididymal CRISP1, is involved in sperm-egg fusion through its binding to complementary sites on the egg surface, supporting the idea of functional cooperation between homologous molecules to ensure the success of fertilization. © 2007 by the Society for the Study of Reproduction, Inc. 2007 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00063363_v76_n4_p701_Busso http://hdl.handle.net/20.500.12110/paper_00063363_v76_n4_p701_Busso |
| 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 Ovum Sperm Testis cell protein isoprotein protein antibody protein CRISP1 protein CRISP2 recombinant protein secretory protein unclassified drug acrosome reaction animal cell animal tissue article cell fusion cell surface cellular distribution controlled study epididymis female fertilization fertilization in vitro human immunofluorescence male mouse nonhuman priority journal protein analysis protein binding protein function protein localization spermatozoon capacitation spermatozoon motility spermatozoon penetration testis vitelline membrane zona pellucida Animals Female Glycoproteins Male Mice Mice, Inbred Strains Sperm-Ovum Interactions Spermatozoa Testis Rattus |
| spellingShingle |
Fertilization Ovum Sperm Testis cell protein isoprotein protein antibody protein CRISP1 protein CRISP2 recombinant protein secretory protein unclassified drug acrosome reaction animal cell animal tissue article cell fusion cell surface cellular distribution controlled study epididymis female fertilization fertilization in vitro human immunofluorescence male mouse nonhuman priority journal protein analysis protein binding protein function protein localization spermatozoon capacitation spermatozoon motility spermatozoon penetration testis vitelline membrane zona pellucida Animals Female Glycoproteins Male Mice Mice, Inbred Strains Sperm-Ovum Interactions Spermatozoa Testis Rattus Evidence for the involvement of testicular protein CRISP2 in mouse sperm-egg fusion |
| topic_facet |
Fertilization Ovum Sperm Testis cell protein isoprotein protein antibody protein CRISP1 protein CRISP2 recombinant protein secretory protein unclassified drug acrosome reaction animal cell animal tissue article cell fusion cell surface cellular distribution controlled study epididymis female fertilization fertilization in vitro human immunofluorescence male mouse nonhuman priority journal protein analysis protein binding protein function protein localization spermatozoon capacitation spermatozoon motility spermatozoon penetration testis vitelline membrane zona pellucida Animals Female Glycoproteins Male Mice Mice, Inbred Strains Sperm-Ovum Interactions Spermatozoa Testis Rattus |
| description |
CRISP2, originally known as Tpx-1, is a cysteine-rich secretory protein specifically expressed in male haploid germ cells. Although likely to be involved in gamete interaction, evidence for a functional role of CRISP2 in fertilization still remains poor. In the present study, we used a mouse model to examine the subcellular localization of CRISP2 in sperm and its involvement in the different stages of fertilization. Results from indirect immunofluorescence and protein extraction experiments indi-cated that mouse CRISP2 is an intraacrosomal component that remains associated with sperm after capacitation and the acrosome reaction (AR). In vitro fertilization assays using zona pellucida-intact mouse eggs showed that an antibody against the protein significantly decreased the percentage of penetrated eggs, with a coincident accumulation of perivitelline sperm. The failure to inhibit zona pellucida penetration excludes a detrimental effect of the antibody on sperm motility or the AR, supporting a specific participation of CRSSP2 at the sperm-egg fusion step. In agreement with this evidence, recombinant mouse CRISP2 (recCRISP2) specifically bound to the fusogenic area of mouse eggs, as previously reported for rat CRISP1, an epididymal protein involved in gamete fusion. In vitro competition investigations showed that incubation of mouse zona-free eggs with a fixed concentration of recCRISP2 and increasing amounts of rat CRISP1 reduced the binding of recCRISP2 to the egg, suggesting that the proteins interact with common complementary sites on the egg surface. Our findings indicate that testicular CRISP2, as observed for epididymal CRISP1, is involved in sperm-egg fusion through its binding to complementary sites on the egg surface, supporting the idea of functional cooperation between homologous molecules to ensure the success of fertilization. © 2007 by the Society for the Study of Reproduction, Inc. |
| title |
Evidence for the involvement of testicular protein CRISP2 in mouse sperm-egg fusion |
| title_short |
Evidence for the involvement of testicular protein CRISP2 in mouse sperm-egg fusion |
| title_full |
Evidence for the involvement of testicular protein CRISP2 in mouse sperm-egg fusion |
| title_fullStr |
Evidence for the involvement of testicular protein CRISP2 in mouse sperm-egg fusion |
| title_full_unstemmed |
Evidence for the involvement of testicular protein CRISP2 in mouse sperm-egg fusion |
| title_sort |
evidence for the involvement of testicular protein crisp2 in mouse sperm-egg fusion |
| publishDate |
2007 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00063363_v76_n4_p701_Busso http://hdl.handle.net/20.500.12110/paper_00063363_v76_n4_p701_Busso |
| _version_ |
1768543589909921792 |