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Sperm plasma membrane receptors for the porcine oocyte plasma membrane

Published online by Cambridge University Press:  05 April 2001

Miguel Betancourt
Affiliation:
Departamento de Ciéncias de la Salud, Universidad Autónoma Metropolitana-Iztapalapa, Mexico
Yvonne Ducolomb
Affiliation:
Departamento de Ciéncias de la Salud, Universidad Autónoma Metropolitana-Iztapalapa, Mexico
Irma Jiménez
Affiliation:
Departamento de Ciéncias de la Salud, Universidad Autónoma Metropolitana-Iztapalapa, Mexico
Eduardo Casas
Affiliation:
Departamento de Ciéncias de la Salud, Universidad Autónoma Metropolitana-Iztapalapa, Mexico
Edmundo Bonilla
Affiliation:
Departamento de Ciéncias de la Salud, Universidad Autónoma Metropolitana-Iztapalapa, Mexico
Trish Berger
Affiliation:
Department of Animal Science, University of California-Davis, California, USA

Abstract

In vitro fertilisation (IVF) was used to assess the ability of solubilised sperm plasma membrane (PM) proteins to inhibit the interaction of intact gametes. This is a first step before evaluating the ability of individual isolated proteins to competitively inhibit sperm-oocyte interaction as part of the process of studying the molecular events of fertilisation. Porcine oocytes were aspirated from ovaries, matured for 48 h in Medium 199, and the zona pellucida (ZP) was removed by exposure to acid Tyrode's solution. ZP-free matured oocytes were exposed to 200–800 μg/ml sperm PM protein for 1 h prior to insemination and during gamete co-incubation. Twenty-four hours after insemination with 5 × 105 capacitated sperm/ml, the oocytes were fixed, stained and examined. Sperm PM protein clearly inhibited IVF in a concentration-dependent manner (r = −0.87). The inhibition index (I50%), representing the sperm PM protein concentration necessary to inhibit IVF to 50% of the control value, was 310 µg/ml. These results demonstrate that solubilised sperm PM protein inhibits the interaction of intact gametes as one might expect for receptor-ligand interactions. Furthermore, the complement of sperm PM proteins appeared maximally effective at a calculated concentration of 690 µm/ml, providing a foundation for further studies with individual proteins.

Type
Research Article
Copyright
1998 Cambridge University Press

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