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Stages leading to and following fusion of sperm and egg plasma membranes

Published online by Cambridge University Press:  26 September 2008

Frank J. Longo ,
Susan Cook ,
David H. McCulloh ,
Pedro I. Ivonnet  and
Edward L. Chambers
Frank J. Longo*
Affiliation:
Department of Anatomy, University of Iowa, and Department of Physiology and Biophysics, University of Miami, USA
Susan Cook
Affiliation:
Department of Anatomy, University of Iowa, and Department of Physiology and Biophysics, University of Miami, USA
David H. McCulloh
Affiliation:
Department of Anatomy, University of Iowa, and Department of Physiology and Biophysics, University of Miami, USA
Pedro I. Ivonnet
Affiliation:
Department of Anatomy, University of Iowa, and Department of Physiology and Biophysics, University of Miami, USA
Edward L. Chambers
Affiliation:
Department of Anatomy, University of Iowa, and Department of Physiology and Biophysics, University of Miami, USA
*
Frank J. Longo, Department of Anatomy, University of Iowa, Iowa City, IA 52242, USA. Tel: (319) 335-7730. Fax: (319) 335-7198.
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Summary

The site of gamete interaction of electrophysiologically recorded Lytechinus variegatus eggs, fixed with osmium tetroxide (O5O4) and/or glutaraldehyde (GTA) at varying intervals after the onset of the increase in membrane conductance induced by an attached sperm, has been examined by high-voltage and conventional transmission electron microscopy. Although GTA and a GTA-O5O4 mixture iduced different electrical responses, specimens prepared with the two fixatives were ultrastructurally similar. In specimens observed within 5 s of the change in conductance, the acrosomal process projected through the vitelline layer and abutted the egg plasma membrane. A conspicuous layer of bindin surrounded the acrosomal process and connected the sperm to the egg's vitelline layer. In a fortuitous specimen fixed within 4 s following the change in conductance, the area of contact between the gamete plasma membranes possessed a trilaminar structure that separated the egg's and sperm's cytoplasms. The morphology of this area of contact was consistent with previously proposed intermediates of membrane fusion. Five to six seconds after the change in conductance, the sperm was connected to the egg via a narrow cytoplasmic bridge that consisted of the former acrosomal process and a projection of the egg cortex. The region of the bridge midway between the fused gametes was encircled by dense material that marked the site of sperm-egg fusion. Gamete interactions in which the activation potential was recorded (unclamped egg) were comparable in time and ultrastructure to events taking place in voltage-clamped eggs except for one major difference. Intact cortical granules (one to three) were observed beneath the tip of the incorporating sperm in unclamped eggs fixed following the onset of the activation potential, whereas all cortical granules dehisced in clamped eggs.

Keywords

Cortical granule exocytosisFertilisationMembrane conductanceMembrane fusionSperm-egg fusionSperm-egg interaction
Type
Article
Information
Zygote , Volume 2 , Issue 4 , November 1994 , pp. 317 - 331
Copyright
Copyright © Cambridge University Press 1994

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