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Assessment of the acrosome reastion in Bufo arenarum Spermatozoa by immunostaining: comparison with other methods

Published online by Cambridge University Press:  26 September 2008

María Laura Martínez  and
Marcelo O. Cabada
María Laura Martínez
Affiliation:
Area Biología, Departamento de Ciencias Biológicas, Facultad de Ciencias Bioqui'micas y Farmacéuticas (UNR), and PROMUBIE (CONICET), Suipacha 531, (2000) Rosario, Argentina.
Marcelo O. Cabada*
Affiliation:
Area Biología, Departamento de Ciencias Biológicas, Facultad de Ciencias Bioqui'micas y Farmacéuticas (UNR), and PROMUBIE (CONICET), Suipacha 531, (2000) Rosario, Argentina.
*
Dr Marcelo O. Cabada, Area Biología, Departamento de Ciencias Biológicas, Facultad de Ciencias Bioquímicas y Farmacéuticas (UNR), and PROMUBIE (CONICET), Suipacha 531, (2000) Rosario, Argentina. Telephone and fax: 0054-41-255277.
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Summary

The acrosome reaction in Bufo arenarum spermatozoa has been visualised only by electron microscopy. Different staining procedures for spermatozoa are described in the present study. By light microscopy the acorsomal status cannot be determined, In some reacted sperm, stained with Coomassie blue, a filament that could be the ‘perforatorium’ was observed, as seen by electron microscopy. Several fluorescent lectins were used, but only FITC-PNA binds to acrosomal proteins. However, the fluorescence observed was weak. Indirect immunofluorescence, with antibodies raised against acrosomal matrix, showed different staining patterns between acrosome-reacted and intact spermatozoa. The technique is specific for evaluating acrosomal status in a large population of spermatozoa. Moreover, immunostaining, in contrast with lectin staining, can be carried out in the presence of glycoconjugates such as oocyte extracellular matrix without interference.

Keywords

Acrosome reactionAcrosomal stainingBufo arenarumFertilisationSpermatozoa

Information

Type
Article
Information
Zygote , Volume 4 , Issue 3 , August 1996 , pp. 181 - 190
Copyright
Copyright © Cambridge University Press 1996

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