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Maintenance of Motility in Mouse Sperm Permeabilized with Streptolysin O¹

^a Center for Research on Reproduction & Women's Health and the Department of Obstetrics and Gynecology, University of Pennsylvania Medical Center, Philadelphia, Pennsylvania 19104-6080

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 [-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.

¹ Supported by NIH P01 HD-06274 (S.B.M. and G.L.G.) and T32 HD-07305 (L.R.J.).

² Authors contributed equally to this work.

³ Correspondence: George L. Gerton, Center for Research on Reproduction and Women's Health, John Morgan Building, Room 306, University of Pennsylvania Medical Center, Philadelphia, PA 19104–6080. FAX: 215 349 5118; gerton{at}mail.med.upenn.edu

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