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Regional differentiation of the sperm surface as studied with 125I-diiodofluorescein isothiocyanate, an impermeant reagent that allows isolation of the labeled components.
Gabel CA
,
Eddy EM
,
Shapiro BM
.
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The regional differentiation of the sperm surface has been studied with the aid of a novel covalent labeling technique that permits concurrent cytological, biochemical, and immunological analyses. For these studies isothiocyanate derivatives of fluorescein (FITC) and diiodofluorescein (IFC) were employed: the latter can be prepared with radioiodine to high specific activity (125IFC) and is an impermeant reagent for the erythrocyte surface. Sperm of sea urchin (Strongylocentrotus purpuratus), medaka )Oryzias latipes), and golden hamster bind the fluorescent chromophores with a nonuniform distribution, most of the fluorescence being associated with the midpiece. The radioactive derivative 125IFC permits an analysis of the proteins that are responsible for most of the binding. Additionally, 125 IFC-labeled sperm are capable of fertilizing eggs, as assessed by autoradiography. That IFC labels the surface of the sperm was inferred from the following: (a) the labeling of the surfaces of other cells by fluorescein isothiocyanate and its derivatives; (b) the agglutination of labeled sperm by antibodies directed against IFC; (c) the use of peroxidase-dependent immunocytochemical reaction using anti-IFC antibodies, with analysis by electron microscopy; and (d) extraction of labeled sea urchin sperm with Triton X-100 under conditions that preferentially solubilize the plasma membrane. The antiserum directed against IFC was used to isolate the labeled surface components from Triton X-100 extracts of whole sperm, by immunoprecipitation, with Staphylococcus-A protein serving as a coprecipitant. The results support previous data showing that the sperm surface is a heterogeneous mosaic of restricted domains, one notable zone being the midpiece, where common molecular properties may be shared by sperm with distinctly different morphologies. In addition, IFC-mediated covalent alteration of specific cell surface proteins may be used to label, to identify, and, with the use of anti-IFC antibodies, to isolate such proteins from other cellular constituents.
Aketa,
Physiological studies on the sperm surface component responsible for sperm-egg bonding in sea urchin fertilization. II. Effect of concanavalin A on the fertilizing capacity of sperm.
1975, Pubmed,
Echinobase
Aketa,
Physiological studies on the sperm surface component responsible for sperm-egg bonding in sea urchin fertilization. II. Effect of concanavalin A on the fertilizing capacity of sperm.
1975,
Pubmed
,
Echinobase
Ames,
Lipids of Salmonella typhimurium and Escherichia coli: structure and metabolism.
1968,
Pubmed
Edelman,
Molecular probes of spermatozoan structures.
1971,
Pubmed
Edidin,
Measurement of membrane protein lateral diffusion in single cells.
1976,
Pubmed
Fellous,
Similar location of an early embryonic antigen on mouse and human spermatozoa.
1974,
Pubmed
Finegold,
Sea urchin egg fertilization studied with a fluorescent probe (ANS).
1974,
Pubmed
,
Echinobase
Friend,
Membrane differentiations in freeze-fractured mammalian sperm.
1974,
Pubmed
Gabel,
[125I]diiodofluorescein isothiocyanate: its synthesis and use as a reagent for labeling proteins and cells to high specific radioactivity.
1978,
Pubmed
Gall,
Chemical and structural analysis of mammalian spermatozoa.
1974,
Pubmed
Geiger,
Protein determination by Lowry's method in the presence of sulfhydryl reagents.
1972,
Pubmed
Gibbons,
Flagellar movement and adenosine triphosphatase activity in sea urchin sperm extracted with triton X-100.
1972,
Pubmed
,
Echinobase
Graham,
The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney: ultrastructural cytochemistry by a new technique.
1966,
Pubmed
Kessler,
Cell membrane antigen isolation with the staphylococcal protein A-antibody adsorbent.
1976,
Pubmed
Kinsey,
Fine structural localization of Concanavalin A binding sites on hamster spermatozoa.
1976,
Pubmed
Koehler,
The mammalian sperm surface: studies with specific labeling techniques.
1978,
Pubmed
Koehler,
Media induced alterations of the membrane associated particles of the guinea pig sperm tail.
1975,
Pubmed
Koehler,
Studies on the distribution of antigenic sites on the surface of rabbit spermatozoa.
1975,
Pubmed
Koo,
Topographical location of H-Y antigen on mouse spermatozoa by immunoelectronmicroscopy.
1973,
Pubmed
Lopatin,
Fluorescein. Hapten and antibody active-site probe.
1971,
Pubmed
March,
A simplified method for cyanogen bromide activation of agarose for affinity chromatography.
1974,
Pubmed
Mellish,
Marking boar spermatozoa with fluorochromes for evaluating spermatozoan transport within gilts.
1970,
Pubmed
Nicolson,
Lectin-binding sites on the plasma membranes of rabbit spermatozoa. Changes in surface receptors during epididymal Maturation and after ejaculation.
1977,
Pubmed
Nicolson,
Mobility and the restriction of mobility of plasma membrane lectin-binding components.
1974,
Pubmed
Overstreet,
Transport, capacitation and fertilizing ability of epididymal spermatozoa.
1974,
Pubmed
Peters,
A microfluorimetric study of translational diffusion in erythrocyte membranes.
1974,
Pubmed
Phillips,
The arrangement of proteins in the human erythrocyte membrane.
1970,
Pubmed
Romrell,
Capping and ultrastructural localization of sperm surface isoantigens during spermatogenesis.
1978,
Pubmed
Shapiro,
Limited proteolysis of some egg surface components is an early event following fertilization of the sea urchin, Strongylocentrotus purpuratus.
1975,
Pubmed
,
Echinobase
Smith,
A uterine fluid-mediated sperm-inhibitory system.
1970,
Pubmed
Yanagimachi,
The distribution of negative surface charges on mammalian spermatozoa.
1972,
Pubmed