Immunolocalization of bovine sperm protease BSp120 by light and electron microscopy during capacitation and the acrosome reaction: Its role in in vitro fertilization
Mammalian fertilization involves various steps in which the participation of specific enzymes has been demonstrated by numerous studies. Acrosin is one of the most widely acrosomal protease in mammalian spermatozoa studied, including bovine; however, other proteases have also been described. A new t...
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2004
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| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_1040452X_v69_n4_p411_Cesari http://hdl.handle.net/20.500.12110/paper_1040452X_v69_n4_p411_Cesari |
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paper:paper_1040452X_v69_n4_p411_Cesari2023-06-08T16:00:45Z Immunolocalization of bovine sperm protease BSp120 by light and electron microscopy during capacitation and the acrosome reaction: Its role in in vitro fertilization Vazquez, Mónica Hebe Mammalian fertilization Membrane sperm protein Serine protease acrosin enzyme antibody polyclonal antibody serine proteinase acrosome reaction animal cell article cattle cell membrane controlled study electron microscopy enzyme activity enzyme analysis enzyme localization fertilization in vitro immunofluorescence test immunolocalization incubation time male microscopy nonhuman oocyte priority journal spermatozoon capacitation spermatozoon motility ultrastructure Acrosome Acrosome Reaction Animals Cattle Enzyme Precursors Fertilization in Vitro Immunohistochemistry Male Peptide Hydrolases Sperm Capacitation Bovinae Cavia porcellus Mammalia Sus scrofa Mammalian fertilization involves various steps in which the participation of specific enzymes has been demonstrated by numerous studies. Acrosin is one of the most widely acrosomal protease in mammalian spermatozoa studied, including bovine; however, other proteases have also been described. A new trypsin-like serine protease named bovine serine protease of 120 kDa (BSp120) and its pre-cursor BSp66 (66 kDa) were identified in bovine spermatozoa. Cytological and ultrastructural immunolocalization studies on BSp120 were performed in live and fixed cells. Immunoflorescence assays with specific polyclonal antibodies revealed localization of BSp120 on the sperm head, with a signal homogeneously distributed over the acrosome resembling a horseshoe. After the acrosome reaction, sperm showed a patchy pattern in the acrosomal cap. Immune electron microscopy analysis indicated that BSp120 is located over the head plasma membrane of capacitated spermatozoa and acrosome reacting spermatozoa. To assess BSp120 function in sperm-oocyte interaction, in vitro fertilization studies were conducted. Oocytes were incubated with spermatozoa pre-treated with anti-BSp120, anti-guinea pig acrosin, and anti-BSp120 plus anti-guinea pig acrosin. Pre-treatment of bovine spermatozoa with antibodies towards each protein did not significantly modify fertilization rates. However, when both anti-acrosin and anti-BSp120 antibodies were simultaneously added, there was a significant decrease in the fertilization rate, suggesting that both enzymes may be required for fertilization. Altogether, the results from the present study described the localization of BSp120 over the acrosome of bovine sperm, and suggest its involvement in fertilization. © 2004 Wiley-Liss, Inc. Fil:Vazquez-Levin, M.H. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2004 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_1040452X_v69_n4_p411_Cesari http://hdl.handle.net/20.500.12110/paper_1040452X_v69_n4_p411_Cesari |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Mammalian fertilization Membrane sperm protein Serine protease acrosin enzyme antibody polyclonal antibody serine proteinase acrosome reaction animal cell article cattle cell membrane controlled study electron microscopy enzyme activity enzyme analysis enzyme localization fertilization in vitro immunofluorescence test immunolocalization incubation time male microscopy nonhuman oocyte priority journal spermatozoon capacitation spermatozoon motility ultrastructure Acrosome Acrosome Reaction Animals Cattle Enzyme Precursors Fertilization in Vitro Immunohistochemistry Male Peptide Hydrolases Sperm Capacitation Bovinae Cavia porcellus Mammalia Sus scrofa |
| spellingShingle |
Mammalian fertilization Membrane sperm protein Serine protease acrosin enzyme antibody polyclonal antibody serine proteinase acrosome reaction animal cell article cattle cell membrane controlled study electron microscopy enzyme activity enzyme analysis enzyme localization fertilization in vitro immunofluorescence test immunolocalization incubation time male microscopy nonhuman oocyte priority journal spermatozoon capacitation spermatozoon motility ultrastructure Acrosome Acrosome Reaction Animals Cattle Enzyme Precursors Fertilization in Vitro Immunohistochemistry Male Peptide Hydrolases Sperm Capacitation Bovinae Cavia porcellus Mammalia Sus scrofa Vazquez, Mónica Hebe Immunolocalization of bovine sperm protease BSp120 by light and electron microscopy during capacitation and the acrosome reaction: Its role in in vitro fertilization |
| topic_facet |
Mammalian fertilization Membrane sperm protein Serine protease acrosin enzyme antibody polyclonal antibody serine proteinase acrosome reaction animal cell article cattle cell membrane controlled study electron microscopy enzyme activity enzyme analysis enzyme localization fertilization in vitro immunofluorescence test immunolocalization incubation time male microscopy nonhuman oocyte priority journal spermatozoon capacitation spermatozoon motility ultrastructure Acrosome Acrosome Reaction Animals Cattle Enzyme Precursors Fertilization in Vitro Immunohistochemistry Male Peptide Hydrolases Sperm Capacitation Bovinae Cavia porcellus Mammalia Sus scrofa |
| description |
Mammalian fertilization involves various steps in which the participation of specific enzymes has been demonstrated by numerous studies. Acrosin is one of the most widely acrosomal protease in mammalian spermatozoa studied, including bovine; however, other proteases have also been described. A new trypsin-like serine protease named bovine serine protease of 120 kDa (BSp120) and its pre-cursor BSp66 (66 kDa) were identified in bovine spermatozoa. Cytological and ultrastructural immunolocalization studies on BSp120 were performed in live and fixed cells. Immunoflorescence assays with specific polyclonal antibodies revealed localization of BSp120 on the sperm head, with a signal homogeneously distributed over the acrosome resembling a horseshoe. After the acrosome reaction, sperm showed a patchy pattern in the acrosomal cap. Immune electron microscopy analysis indicated that BSp120 is located over the head plasma membrane of capacitated spermatozoa and acrosome reacting spermatozoa. To assess BSp120 function in sperm-oocyte interaction, in vitro fertilization studies were conducted. Oocytes were incubated with spermatozoa pre-treated with anti-BSp120, anti-guinea pig acrosin, and anti-BSp120 plus anti-guinea pig acrosin. Pre-treatment of bovine spermatozoa with antibodies towards each protein did not significantly modify fertilization rates. However, when both anti-acrosin and anti-BSp120 antibodies were simultaneously added, there was a significant decrease in the fertilization rate, suggesting that both enzymes may be required for fertilization. Altogether, the results from the present study described the localization of BSp120 over the acrosome of bovine sperm, and suggest its involvement in fertilization. © 2004 Wiley-Liss, Inc. |
| author |
Vazquez, Mónica Hebe |
| author_facet |
Vazquez, Mónica Hebe |
| author_sort |
Vazquez, Mónica Hebe |
| title |
Immunolocalization of bovine sperm protease BSp120 by light and electron microscopy during capacitation and the acrosome reaction: Its role in in vitro fertilization |
| title_short |
Immunolocalization of bovine sperm protease BSp120 by light and electron microscopy during capacitation and the acrosome reaction: Its role in in vitro fertilization |
| title_full |
Immunolocalization of bovine sperm protease BSp120 by light and electron microscopy during capacitation and the acrosome reaction: Its role in in vitro fertilization |
| title_fullStr |
Immunolocalization of bovine sperm protease BSp120 by light and electron microscopy during capacitation and the acrosome reaction: Its role in in vitro fertilization |
| title_full_unstemmed |
Immunolocalization of bovine sperm protease BSp120 by light and electron microscopy during capacitation and the acrosome reaction: Its role in in vitro fertilization |
| title_sort |
immunolocalization of bovine sperm protease bsp120 by light and electron microscopy during capacitation and the acrosome reaction: its role in in vitro fertilization |
| publishDate |
2004 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_1040452X_v69_n4_p411_Cesari http://hdl.handle.net/20.500.12110/paper_1040452X_v69_n4_p411_Cesari |
| work_keys_str_mv |
AT vazquezmonicahebe immunolocalizationofbovinespermproteasebsp120bylightandelectronmicroscopyduringcapacitationandtheacrosomereactionitsroleininvitrofertilization |
| _version_ |
1768545703183777792 |