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Digitale Medien

Visualization and characterization of the acrosome reaction of human spermatozoa by immunolocalization with monoclonal antibody (1987)

Moore, H. D. M. ; Smith, C. A. ; Hartman, T. D. ; [weitere]

New York, NY : Wiley-Blackwell

Gamete Research 17 (1987), S. 245-259

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ISSN: 0148-7280

Schlagwort(e): hamster ova ; epi-fluorescence ; electron microscopy ; Life and Medical Sciences ; Cell & Developmental Biology

Quelle: Wiley InterScience Backfile Collection 1832-2000

Thema: Biologie

Notizen: A monoclonal antibody generated against hamster epididymal spermatozoa and recognizing an antigen within the acrosome was used in conjunction with FITC-antimouse immunoglobulin as a marker of the human acrosome during sperm development, capacitation, and the acrosome reaction. The specificity of binding of the monoclonal antibody was assessed using immunolocalization by epi-fluorescence and electron microscopy. Immunofluorescence revealed that antibody bound over the entire anterior acrosome in hamster and human spermatozoa. Ultrastructural localization indicated that antigen was predominantly present on the inner face of the outer acrosomal membrane and within the acrosomal content. Qualitative specificity was studied using a highly purified preparation of hamster acrosomes in an enzyme-linked immunosorbent assay. Since the antibody rapidly visualized human acrosomes, it was used to detect abnormal acrosome morphology of mature spermatozoa and to mark spermatids present in the ejaculate. During incubation in capacitating medium, changes in the immunofluorescence of live or methanol fixed spermatozoa were correlated with incubation interval and the ability of spermatozoa to fuse with zona-free hamster oocytes. Spermatozoa bound to zona-free hamster oocytes displayed no fluorescence, confirming that acrosome loss occurred before spermatozoa attached to the vitellus.

Zusätzliches Material: 14 Ill.

Materialart: Digitale Medien

URL: http://dx.doi.org/10.1002/mrd.1120170308

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Digitale Medien

Separation of round spermatids from the rat using an immunoselection panning technique (1995)

Pelengaris, S. A. ; Moore, H. D. M.

New York, NY [u.a.] : Wiley-Blackwell

Molecular Reproduction and Development 41 (1995), S. 348-354

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ISSN: 1040-452X

Schlagwort(e): Spermatogenesis ; Germ cells ; Immunoselection ; Monoclonal antibody ; RNA ; Life and Medical Sciences ; Cell & Developmental Biology

Quelle: Wiley InterScience Backfile Collection 1832-2000

Thema: Biologie

Notizen: A method was devised for the isolation of round spermatids from the rat using a positive immunoselection technique (panning). A testis suspension was prepared from adult rats by enzymatic digestion of seminiferous tubules with collagenase. Specific mouse monoclonal antibody (97.25) was indirectly attached to Petri dishes and used in a panning protocol to purify spermatids from the testis cell suspension. The quantity and purity of cells isolated were determined by cell counts and histochemical (periodic acid-Schiff stain) or by immunostaining with acrosome-specific antibodies. A mean yield of 1.38 ± 0.15 × 107 cells per dish was obtained with a purity of more than 90%. The viability of the cells was confirmed by epifluorescent microscopy with propidium iodide/carboxyfluorescein acetate probes. Northern blot analysis of RNA extracted directly from the dish indicated good integrity of a spermatid-specific transcript of glyceraldehyde-3-phosphate dehydrogenase (GAPDH). © 1995 Wiley-Liss, Inc.

Zusätzliches Material: 5 Ill.

Materialart: Digitale Medien

URL: http://dx.doi.org/10.1002/mrd.1080410310

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Digitale Medien

Acrosome reaction of stallion spermatozoa evaluated with monoclonal antibody and zona-free hamster eggs (1990)

Zhang, J. ; Boyle, M. S. ; Smith, C. A. ; [weitere]

New York, NY [u.a.] : Wiley-Blackwell

Molecular Reproduction and Development 27 (1990), S. 152-158

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ISSN: 1040-452X

Schlagwort(e): Stallion spermatozoa ; Acrosome reaction ; Monoclonal antibody ; Zona-free hamster eggs ; Life and Medical Sciences ; Cell & Developmental Biology

Quelle: Wiley InterScience Backfile Collection 1832-2000

Thema: Biologie

Notizen: The acrosome of the stallion spermatozoon was visualized by indirect immunofluorescence with monoclonal antibody (18.6) which recognized an integral acrosomal membrane component. Localization was confirmed by electron microscopy using peroxidase labelled antibody. In fresh semen samples (n = 19), 73.9 ± 9.1% of the spermatozoa from five fertile stallions displayed a uniform bright fluorescence over their acrosome region. In two semen samples from an infertile stallion only 28% and 35% of spermatozoa showed the same pattern of fluorescence. Spermatozoa from fertile stallions incubated for up to 12 hours in TALP medium maintained motility and exhibited a significant progressive loss of acrosomes as detected by immunofluorescence. Alternatively, a similar loss of acrosomes could be induced with calcium ionophore A23187 over a 90 minute incubation. Ultrastructural observations and incubation with zona-free hamster eggs indicated that only with ionophore treatment was immunofluorescent acrosome loss correlated with a physiological acrosome reaction, while prolonged sperm incubation led to degenerative membrane changes. It was concluded that, if carefully validated, immunofluorescent localization of the acrosome of stallion sperm with monoclonal antibody could be used to monitor the acrosome reaction. Furthermore, definitive acrosome visualization would be valuable in assessing semen quality.

Zusätzliches Material: 6 Ill.

Materialart: Digitale Medien

URL: http://dx.doi.org/10.1002/mrd.1080270210

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