Maintenance of motility in mouse sperm permeabilized with streptolysin O.
Biol Reprod
; 60(3): 683-90, 1999 Mar.
Article
en En
| MEDLINE
| ID: mdl-10026116
ABSTRACT
One approach to studying the mechanisms governing sperm motility is to permeabilize sperm and examine the regulation of motility by manipulating the intracellular milieu of the cell. The most common method of sperm permeabilization, detergent treatment, has the disadvantage that the membranes and many proteins are extracted from the cell. To avoid this problem, we have developed a method that uses streptolysin O to create stable pores within the plasma membrane while leaving internal membranes intact. Sperm were permeabilized, preincubated, and then treated with 0.6 U/ml of streptolysin O. Permeabilization was assessed by fluorescent dye technologies and endogenous protein phosphorylation using exogenously added [gamma-32P]ATP. Streptolysin O-induced permeabilization rendered the sperm immotile, and the effect was Ca2+-dependent. When the cells were treated simultaneously with a medium containing ATP, streptolysin O-treated sperm maintained flagellar movement. These results demonstrate that the streptolysin O permeabilization model system is a useful experimental method for studying the mechanisms that regulate sperm motility since it allows the flagellar apparatus to be exposed to various exogenously added molecules.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Motilidad Espermática
/
Estreptolisinas
/
Permeabilidad de la Membrana Celular
Tipo de estudio:
Prognostic_studies
Límite:
Animals
Idioma:
En
Revista:
Biol Reprod
Año:
1999
Tipo del documento:
Article
País de afiliación:
Estados Unidos