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The kinetics of human sperm binding to the human zona pellucida and zona-free hamster oocyte in vitro (1985)

Singer, Sherry L. ; Lambert, Hovey ; Overstreet, James W. ; [et al.]

New York, NY : Wiley-Blackwell

Gamete Research 12 (1985), S. 29-39

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ISSN: 0148-7280

Keywords: human ; sperm ; zona pellucida ; zona-free hamster oocyte ; Life and Medical Sciences ; Cell & Developmental Biology

Source: Wiley InterScience Backfile Collection 1832-2000

Topics: Biology

Notes: In these experiments the mean number of sperm bound to the zonae pellucidae of immature human oocytes and to the vitelli of zona-free hamster oocytes were counted after centrifuging the oocytes through a discontinuous dextran gradient into a fixative. Sperm suspensions (107 sperm/ml) were preincubated in BWW medium containing 35 mg/ml of human serum albumin for 0 to 20 hr and coincubated with oocytes for 4 hr.The kinetics of sperm binding and sperm penetration were clearly different for the two types of oocytes in this system. Sperm binding to the zona pellucida appeared to be associated with zona penetration; when the zona was penetrated, many sperm were tightly bound and vice versa. A similar association between human sperm binding to the zonafree hamster oocyte and hamster oocyte penetration was not so apparent. Transmission electron microscopy revealed that all sperm which were “bound” to the hamster vitellus were acrosome reacted. On the surface of the human zona pellucida acrosome intact and acrosome reacted sperm were observed.These results suggest that tight binding of spermatozoa to the zona pellucida may be an important preliminary step in human fertilization. If this is true the activity of the sperm's “receptor for zona” may not be detected by the assay with zona-free hamster oocytes.

Additional Material: 3 Ill.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1002/mrd.1120120104

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Interaction of human spermatozoa with the human zona pellucida and zona-free hamster oocyte following capacitation by exposure to human cervical mucus (1985)

Gould, John E. ; Overstreet, James W. ; Hanson, Frederick W.

New York, NY : Wiley-Blackwell

Gamete Research 12 (1985), S. 47-54

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ISSN: 0148-7280

Keywords: sperm ; human ; zona pellucida ; cervical mucus ; zona-free hamster oocyte ; Life and Medical Sciences ; Cell & Developmental Biology

Source: Wiley InterScience Backfile Collection 1832-2000

Topics: Biology

Notes: The functional capacity for sperm interaction with the human zona pellucida and zona-free hamster oocyte was tested after human spermatozoa were capacitated by passage through a column of human cervical mucus. The results were compared with those obtained when spermatozoa from an aliquot of the same semen sample were capacitated by the standard laboratory methods involving sequential washing by dilution and centrifugation of the semen. Washed-capacitated sperm suspensions were more successful than mucus-capacitated sperm in attaching to the zona-free hamster oocyte and in fusing with its oolemma. However, the ability of mucus-capacitated sperm to penetrate the human zona pellucida was equal to washed capacitated sperm. These experiments demonstrate an approach that may be useful in comparative studies of human sperm capacitation in vivo and in vitro.

Additional Material: 1 Ill.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1002/mrd.1120120106

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Two simple methods for detecting acrosome-reacted human sperm (1986)

Cross, Nicholas L. ; Morales, Patricio ; Overstreet, James W. ; [et al.]

New York, NY : Wiley-Blackwell

Gamete Research 15 (1986), S. 213-226

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ISSN: 0148-7280

Keywords: acrosome reaction ; lectin ; spermatozoa ; Life and Medical Sciences ; Cell & Developmental Biology

Source: Wiley InterScience Backfile Collection 1832-2000

Topics: Biology

Notes: We describe two methods for detecting acrosome reactions of human sperm at the light microscopic level. The techniques include the use of a supravital stain to detect dead sperm in order to differentiate between “physiological” and “degenerative” acrosome reactions. Sperm are incubated with the supravital stain Hoechst 33258 (a fluorescent DNA-binding dye with limited membrane permeability), washed, suspended in 95% ethanol for fixation and permeabilization, and dried onto slides. The sperm are then labeled either by indirect immunofluorescence using rabbit anti-human sperm antiserum or with fluoresceinated Pisum sativum agglutinin (PSA). Both probes intensely label the acrosomal region of acrosome-intact sperm. Electron microscopy revealed the major site of PSA binding to be the acrosomal contents. Acrosome-reacted sperm have diminished acrosomal labeling by both probes; sperm with nuclei labeled by Hoechst stain are considered nonviable, and are excluded from the assay. Both assays are rapid, give similar results, and detect an increase in acrosome reactions following exposure to the ionophore A23187.

Additional Material: 10 Ill.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1002/mrd.1120150303

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