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BOR - Papers in Press, published online ahead of print October 23, 2002.
Biol Reprod 2002, 10.1095/biolreprod.102.009175
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biolreprod.102.009175v1
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BIOLOGY OF REPRODUCTION 68, 664–672 (2003)
DOI: 10.1095/biolreprod.102.009175
© 2003 by the Society for the Study of Reproduction, Inc.


Testis

Real-Time Observations of Individual Macaque Sperm Undergoing Tight Binding and the Acrosome Reaction on the Zona Pellucida1

Theodore L. Tollnera, Ashley I. Yudina, Gary N. Cherrb, and James W. Overstreet2,a

a Division of Reproductive Biology, b Department of Obstetrics and Gynecology, and Bodega Marine Laboratory, University of California, Davis, California 95616

Changes in binding affinity, acrosomal status, and motility of living sperm on the zona pellucida were for the first time in any mammalian species directly observed and analyzed with video microscopy. A single zona was air-dried and rehydrated on a microscope slide, and a coverslip supported by glass beads was added. Capacitated sperm were added together with Alexa-SBTI, a probe for acrosin that can detect the acrosome reaction. The heads of loosely attached sperm oscillated on the zona and the flagella beat symmetrically with a sigmoid-shaped waveform. Tight binding was observed after 16 sec as the sperm head became fixed in place on the zona. The shape of the flagellar beat simultaneously shifted to a more rigid, C-shaped waveform. The first signs of the acrosome reaction were detected within 11 sec of tight binding. Rapid flushing removed approximately 65% of sperm that were loosely attached but only 2% of those that were tightly bound. In the 2 min following the onset of tight binding, the lateral displacement of the flagellum increased by approximately 30% and the beat frequency decreased by 25%. Lignosulfonic acid (LSA) inhibited loose sperm attachment and the development of tight binding. LSA had no effect on the time of the acrosome reaction following tight binding or on changes in motility that followed tight binding. These data suggest that LSA affects the initial attachment or docking of sperm to the zona, a step that may align or recruit one or more specific zona receptors to be responsible for mediating the acrosome reaction.

1 This work is supported by grants U54-HD29125 and P51-RR00169 from the National Institutes of Health and by the Andrew W. Mellon Foundation.

2 Correspondence: James W. Overstreet, Center for Health and the Environment, One Shields Avenue, University of California, Davis, CA 95616. FAX: 530 752 5300; jwoverstreet{at}ucdavis.edu




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