Immunolocalization of the sea urchin sperm receptor in eggs and maturing ovaries.
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 at the height of the spawning season, and present only on the cell surface of mature oocytes in the lumen of the ovaries. Our results indicate that receptor is synthesized early in oogenesis and is rapidly moved to the egg cell surface.
Ruiz-Bravo
N
N
Department of Biochemistry and Molecular Biology, University of Texas, MD Anderson Cancer Center, Houston 77030.
Janak
D J
DJ
Lennarz
W J
WJ
D000818:Animals; D002499:Centrifugation, Density Gradient; D000939:Epitopes / Q000032:analysis; D005260:Female; D005306:Fertilization; D005455:Fluorescent Antibody Technique; D006023:Glycoproteins / Q000032:analysis*; D008297:Male; D009865:Oocytes / Q000032:analysis*; D009866:Oogenesis; D010053:Ovary / Q000032:analysis*; D011956:Receptors, Cell Surface / Q000032:analysis* / Q000302:isolation & purification; D012617:Sea Urchins; D013045:Species Specificity; D013084:Sperm-Ovum Interactions
D000939:Epitopes; D006023:Glycoproteins; D011956:Receptors, Cell Surface; C035027:egg surface sperm receptor