Structural properties of the membrane of intact human spermotozoa

Mercado, Efraı́n; Rosado, Adolfo

doi:10.1016/0005-2736(73)90080-1

Cited by 25 publications

(16 citation statements)

References 15 publications

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“…Fluorescence measurements were made with a recording spectrofluorometer (Perkin-Elmer MPF-2A) at 475 nm using the procedures described by Mercado & Rosado (1973) (Text- fig. 1) with ANS to a maximum concentration of 100 pM-ANS (Datta & Penefsky, 1970).…”

Section: Fluorometrymentioning

confidence: 99%

“…1) with ANS to a maximum concentration of 100 pM-ANS (Datta & Penefsky, 1970). The sample volume was chosen to limit any increase in volume to less than 5% (Mercado & Rosado, 1973), thereby minimizing its effect on the level of fluorescence.…”

Section: Fluorometrymentioning

confidence: 99%

“…3a) or protein concentration ) and each line was extrapolated to the ordinate by regression analysis. The maximum fluorescence at each ANS concentration was calculated as the reciprocal of the ordinate intercept (Brocklehurst, Freedman, Hancock & Radda, 1970 (Scatchard, 1949;Datta & Penefsky, 1970;Mercado & Rosado, 1973;Aithal et ai, 1974) for each sperm or seminal plasma protein concentration (Text- fig. 4).…”

Section: Fluorometrymentioning

confidence: 99%

“…KD was determined as the negative of the value calculated for the gradient and as the ordinate intercept (Table 1). Alternatively, the reciprocal of the concentration of ANS bound was plotted against the reciprocal of the concentration of unbound dye (Klotz, Walker & Pivan, 1946;Laurence, 1952;Weber & Young, 1964a, b;Mercado & Rosado, 1973 (Text-fig. 7).…”

Section: Fluorometrymentioning

confidence: 99%

“…The dye 1-anilino-naphthalene-8-sulphonate (ANS) fluoresces strongly when adsorbed to proteins in solution (Weber & Laurence, 1954) and when bound to human spermatozoa (Mercado & Rosado, 1973). Microscopical studies have indicated that the fluorescence resulting from ANS interaction with ram spermatozoa is reduced by prior incubation of spermatozoa with egg yolk (Watson, 1975).…”

Section: Introductionmentioning

confidence: 99%

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A spectrofluorometric investigation, using 1-anilino-naphthalene-8-sulphonate, of the interaction between washed bovine spermatozoa and seminal plasma or egg-yolk lipoprotein

MacDonald¹,

Foulkes²

1981

Reproduction

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Summary. The dye 1-anilino-naphthalene-8-sulphonate (ANS) bound at 94\m=.\5\ m=+-\ 2\m=.\7 pmol per 106 spermatozoa (Kd = 1\m=.\83\ m=+-\0\m=.\13\m=x\ 10\m=-\5 m) and 7\m=.\30\ m=+-\ 0\m=.\17 pmol per mg seminal plasma protein (Kd = 2\m=.\19\ m=+-\0\m=.\06\m=x\10\m=-\5 m). Equilibration of excess ANS with a mixture of washed spermatozoa and seminal plasma resulted in a significant increase in fluorescence over that calculated by summation of the individual levels (P < 0\m=.\05). A less pronounced but significant increase was seen when an egg-yolk lipoprotein, previously shown to have cryoprotective properties, was similarly added to washed spermatozoa (P < 0\m=.\05).This increase in fluorescence was not reduced by washing spermatozoa\p=n-\lipoproteinmixtures, suggesting that an interaction between the lipoprotein and the cell membrane had occurred.

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Section: Fluorometrymentioning

confidence: 99%

Section: Fluorometrymentioning

confidence: 99%

Section: Fluorometrymentioning

confidence: 99%

Section: Fluorometrymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

A spectrofluorometric investigation, using 1-anilino-naphthalene-8-sulphonate, of the interaction between washed bovine spermatozoa and seminal plasma or egg-yolk lipoprotein

MacDonald¹,

Foulkes²

1981

Reproduction

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Biochemical studies of taste sensation. III. Preparation of a suspension of bovine taste bud cells and their labeling with a fluorescent probe

Brand

Çağan

1976

J. Neurobiol.

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A method to prepare suspensions of taste bud cells is described. Bovine circumvallate papillae, which contain most of the taste buds in this animal, are incubated in collagenase-containing medium and the epidermal sidewall tissue is then dissected from the inner gelatinous dermis. The sidewall tissue, which contains the taste buds, is gently homogenized by manual operation of an all-glass homogenizer with a loose-fitting pestle. The suspended material is separated on a discontinous Ficoll gradient (2%, 8%, 10%, 12% w/w). The material banding at the 8-2% interface is greatly enriched in spindle-shaped cells that are morphologically similar to taste bud cells as they appear in situ. These cells are not seen when the procedure is done with tissues devoid of taste buds, namely the upper surface of the circumvallate papilla or epithelium from the intermolar eminence. Fluorescence analysis indicates that the hydrophobic probe, 8-anilino-1-naphthalenesulfonate (ANS), binds to relatively nonpolar sites in the suspension. It is postulated that the probe is adsorbing onto the surface membrane of the cell. These preparations may be useful in studying specificity and transduction in taste sensation.

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Membrane organization and differentiation in the guinea-pig spermatozoon

Friend

1984

Ultrastructure of Reproduction

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Structural properties of the membrane of intact human spermotozoa

Cited by 25 publications

References 15 publications

A spectrofluorometric investigation, using 1-anilino-naphthalene-8-sulphonate, of the interaction between washed bovine spermatozoa and seminal plasma or egg-yolk lipoprotein

A spectrofluorometric investigation, using 1-anilino-naphthalene-8-sulphonate, of the interaction between washed bovine spermatozoa and seminal plasma or egg-yolk lipoprotein

Biochemical studies of taste sensation. III. Preparation of a suspension of bovine taste bud cells and their labeling with a fluorescent probe

Membrane organization and differentiation in the guinea-pig spermatozoon

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