Mol. Hum. Reprod. Advance Access originally published online on June 16, 2007
Molecular Human Reproduction 2007 13(8):557-565; doi:10.1093/molehr/gam042
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Calcium and sperm components in the establishment of the membrane block to polyspermy: studies of ICSI and activation with sperm factor
1 Division of Reproductive Biology, Department of Biochemistry and Molecular Biology, Johns Hopkins University, Bloomberg School of Public Health, Room W3606, 615 North Wolfe Street, Baltimore, MD 21205, USA 2 Department of Veterinary and Animal Sciences, University of Massachusetts, Amherst, MA, USA 3Present address: Shire Pharmaceuticals, San Francisco, CA and Wayne, PA, USA 4Present address: Department of Pharmacology and Cancer Biology, Duke University Medical Center, Durham, NC, USA
5 Correspondence address. Tel: +1-410-614-5557; Fax: +1-410-614-2356; E-mail: jpevans{at}jhsph.edu
One important result of egg activation is the establishment of blocks to prevent polyspermic fertilization; these blocks are established on the zona pellucida and the egg plasma membrane. This study examines what the sperm brings to the egg to induce the establishment of the membrane block to polyspermy, building on past evidence that membrane block establishment does not occur in response to parthenogenetic stimuli that induce a single transient increase in cytosolic Ca2+ or intracytoplasmic sperm injection (ICSI). We test the hypotheses that (i) sperm-associated Ca2+ release activity triggers membrane block establishment; (ii) introduction of sperm contents via variations on ICSI protocols (resulting in improved Ca2+ transients, egg activation and embryo development over traditional ICSI protocols) triggers membrane block establishment and (iii) sperm adhesion [binding of an extracellular sperm ligand(s) to an egg receptor(s)] combined with sperm-associated Ca2+ release activity triggers membrane block establishment. Interestingly, none of these stimuli induced establishment of the membrane block to polyspermy in mouse eggs. However, the sperm-associated remodeling of the egg cortical cytoskeleton differs between conventionally fertilized and ICSI-fertilized eggs; taken with our previous data implicating actin microfilaments in membrane block establishment, this raises the possibility that cortical reorganization may be a contributing factor. In sum, fertilization-like Ca2+ transients, either alone or combined with sperm–egg binding, are not sufficient for membrane block establishment, but that an event(s) associated with gamete interaction plays a role in this membrane function change.
Key words: fertilization/ICSI/egg activation/polyspermy/calcium
Submitted on April 18, 2007; resubmitted on May 3, 2007; accepted on May 10, 2007.