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  • 1
    ISSN: 1040-452X
    Keywords: Macaca fasicularis ; Semen ; Capacitation ; Zona binding ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Capacitation of macaque sperm with caffeine and dbcAMP is required for fertilization in vitro. This study determined the separate effects of caffeine and dbcAMP on sperm-zona pellucida binding and the acrosome reaction of zona bound sperm. Semen from 6 cynomolgus macaques was washed through 60% Percoll, resuspended, and washed with BWW media and incubated for 2.5 hr. Caffeine, dbcAMP (2 mM each), or both (1 mM each) were added to aliquots of the sperm suspensions. Immature macaque oocytes were placed into drops of sperm suspensions, coincubated with sperm for 30 sec, and either fixed immediately or removed to sperm-free media and incubated 1 hr before fixation. There were no significant diffences between groups in the percentage of live, acrosome-reacted sperm in suspension. Treatment with caffeine and dbcAMP or with caffeine alone, significantly increased the number of sperm bound to each zona pellucida (96 ± 16 and 81 ± 17, respectively) compared to control and dbcAMP treatment (15 ± 4 and 28 ± 13). However, treatment with dbcAMP, alone and with caffeine, resulted in a higher percentage of acrosome-reacted sperm on the zona (15.2 ± 2.1 and 9.0 ± 0.6) than control or caffeine treatment (3.0 ± 1.4 and 2.4 ± 0.5). Effects on sperm motility consistent with hyperactivation were detected only when both caffeine and dbcAMP were present. Although both caffeine and dbcAMP are presumed to increase or to produce the same effects as increased intracellular cAMP levels, these compounds have different effects on the ability of sperm to bind to the zona and to undergo the acrosome reaction. © 1994 Wiley-Liss, Inc.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 0148-7280
    Keywords: lectins ; sperm ; capacitation ; plasma membrane ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Human sperm incubated in vitro in BWW medium containing 35 mg/ml human serum albumin acquire the capacity to penetrate the human zona pellucida and to fuse with the zona-free hamster oocyte. We have studied changes in lectin-induced agglutination of human sperm during incubation in this medium to detect alterations in the sperm surface which may be correlated with the acquisition of these functions. Sperm incubated for 1, 6, or 24 hr were combined with two-fold dilutions of lectins for 30 min at 37°C, in 5% CO2, balance air. When pooled data from five donors were analyzed, the average sperm agglutination titer of wheat germ agglutinin (WGA), phytohemagglutinin-E (PHA), Lens culinaris agglutinin (LCA), peanut agglutinin (PNA), and Pisum sativum agglutinin (PSA) was found to increase significantly (P ≤ 0.06) with incubation in vitro, although there was considerable variation between ejaculates. Ulex europaeus and Dolichos biflorus agglutinins did not agglutinate human sperm (≤250 μg/ml). Results of this screening demonstrate the alteration of sperm surface components during in vitro incubation and suggest that WGA, PHA, LCA, and PSA may prove useful in efforts to correlate changes in the sperm surface with the ability of the sperm to fertilize the egg.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Gamete Research 22 (1989), S. 443-469 
    ISSN: 0148-7280
    Keywords: sperm ; transport ; mucus ; cumulus ; zona pellucida ; fertilization ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Mechanisms of mammalian sperm migration through the female reproductive tract and ovum vestments are described. The perspective is biophysical as well as biochemical and morphological, and the focus is upon the role of sperm motility in these processes. Sperm forward progression is characterized as an interactive process between the the cell and its environment, and the mediation of flagellar bend propagation by the physical properties of its surroundings is described. These properties, together with flagellar beat kinematics, sperm morphology, and surface properties, determine the magnitude of the forces generated by sperm and their consequent rate of progression. Sperm interactions with the cervical mucus, the cumulus oophorus, and the zona pellucida are described. The poorly understood affinity of the sperm surface for the macromolecules of the mucus, cumulus, and zona is stressed, as is the viscoelastic structural mechanical resistance of these biopolymers to sperm motion. The kinematics and consequences of hyperactivated sperm motion are presented, with emphasis on objective characterization of such motion (as a biomarker), along with analysis of the mechanical advantage that such motion may confer on spermatozoa during egg-vestment interaction.
    Additional Material: 3 Ill.
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  • 4
    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.
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  • 5
    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
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