Aug 1, 1989

Immunolocalization of the sea urchin sperm receptor in eggs and maturing ovaries

Biology of Reproduction
N Ruiz-BravoW J Lennarz

Abstract

The sperm receptor from Strongylocentrotus purpuratus eggs is a high molecular weight proteoglycan-like molecule that inhibits fertilization species-specifically in a competition bioassay. A preparation of highly active sperm receptor that contained one major N-terminal sequence was used to generate polyclonal antibody in rabbits. This antibody species-specifically inhibited fertilization at low concentrations without interfering with fertilization envelope elevation. On immunofluorescence microscopy, the antibody recognized determinants on the mature egg cell surface and in the cortical granules just beneath the surface. Preabsorption of the antibody with the calcium-soluble fraction of the exudate from cortical granules rendered the antibody specific for the cell surface in mature eggs and still able to inhibit fertilization at the same concentrations as before treatment with cortical granule exudate. With antibody preabsorbed with cortical granule and by counting antibody-gold particles viewed by electron microscopy, sperm receptor was almost undetectable on the cell surface of immature oocytes in preseason ovaries, present on the cell surface and intracellularly in immature oocytes of ovaries collected at the beginning or a...Continue Reading

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Mentioned in this Paper

Fertility
Centrifugation, Density Gradient
Immunofluorescence Assay
Calcium [EPC]
ZP3 gene
Ovum
Entire Lumen of Body System
Calcium
Proteoglycan
Cortical Granule

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