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  • 1
    Electronic Resource
    Electronic Resource
    Assessment of Pisum sativum agglutinin in identifying acrosomal damage in stallion spermatozoa (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 32 (1992), S. 23-27 
    Details
    ISSN: 1040-452X
    Keywords: Assay ; Lectin ; binding ; Fluorescence ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The use of fluorescein-conjugated Pisum sativum agglutinin (FITC-PSA) was evaluated for its ability to distinguish acrosome-intact from acrosome-damaged stallion spermatozoa. Incubation of fresh (acrosomeintact) and frozen-thawed (acrosome-damaged) spermatozoa with FITC-PSA resulted in acrosome-intact spermatozoa that exhibited no fluorescence, while acrosome-damaged spermatozoa exhibited fluorescent staining over the rostral portion of the head and equatorial segment. Experiments using mixtures of various ratios of acrosome-intact and acrosome-damaged spermatozoa determined the precision (intrasample coefficient of variation), and linearity (increased percentage of spermatozoa with PSA binding, with increased percentage of frozenthawed spermatozoa in a sample) of FITC-PSA binding. The binding of FITC-PSA increased in samples as the portion of frozen-thawed (acrosome-damaged) to fresh (acrosomeintact) spermatozoa increased. A positive correlation existed (r = 0.98, P 〈 0.05) between the percentage of FITC-PSA bound sperm and the proportion of damaged spermatozoa added to a sample. Location of PSA lectin binding on acrosome-damaged spermatozoa, determined by electron microscopy using gold-conjugated PSA, was to components of the outer acrosomal membrane and acrosomal matrix. These results demonstrate that FITC-PSA binding may be useful in determining acrosomal integrity of fresh and frozen-thawed stallion spermatozoa. © 1992 Wiley-Liss, Inc.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/mrd.1080320105
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  • 2
    Electronic Resource
    Electronic Resource
    Dilauroylphosphatidylcholine liposome effects on the acrosome reaction and in vitro penetration of zona-free hamster eggs by bull sperm: I. A fertility assay for fresh semen (1987)
    New York, NY : Wiley-Blackwell
    Gamete Research 16 (1987), S. 133-145 
    Details
    ISSN: 0148-7280
    Keywords: liposomes ; spermatozoa ; egg penetration ; bull fertility ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Fresh sperm from five bulls having nonreturn rates ranging from 48% to 77% were treated with 15.7, 21.0, 26.2, 31.5, 36.7, and 42.0 μM dilauroylphosphatidylcholine (PC12) to induce the sperm acrosome reaction (AR). Treated sperm were incubated 3 hr with zona-free hamster eggs at 39°C prior to fixation. The eggs were then stained and examined for sperm penetration. Differences in the percentages of motile sperm and of sperm exhibiting an AR among bulls were small when compared on a within-liposome-concentration basis. Increasing the PC12 concentration from 15.7 μM to 42.0 μM increased the percentage of sperm exhibiting an AR for all bulls. At the lowest lipid concentration (15.7 μM), the percentage of eggs penetrated by sperm from the five bulls was 6% to 36%, with 0% in controls. When sperm were incubated with increasing lipid concentrations, the egg penetration rate increased to over 80%, and the total number of sperm increased to over 100 per 36 eggs in each treatment for every bull. These penetration rates decreased at the highest lipid concentration. A correlation between the PC12 concentration maximizing egg penetration and the nonreturn rate of -.63 was found. The correlation between the PC12 concentration maximizing the total number of penetrated sperm per treatment and the bull nonreturn rate was -.96. It was concluded that PC 12 liposomes induce the AR in bull spermatozoa, which enables them to penetrate zona-free hamster eggs. High fertility bulls required less lipid to induce the AR than did lower fertility bulls. Consequently, this assay of fresh semen could provide a laboratory method to estimate the fertility of a bull.
    Additional Material: 1 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/mrd.1120160205
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  • 3
    Electronic Resource
    Electronic Resource
    Dilauroylphosphatidylcholine liposome effects on the acrosome reaction and in vitro penetration of zona-free hamster eggs by bull sperm: II. A fertility assay for frozen-thawed semen (1987)
    New York, NY : Wiley-Blackwell
    Gamete Research 16 (1987), S. 147-158 
    Details
    ISSN: 0148-7280
    Keywords: egg penetration ; bull fertility ; motile sperm ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Frozen-thawed sperm from five bulls with fertility rates ranging from 48% to 77% were treated with seven concentrations of dilauroylphosphatidylcholine (PC12) liposomes to induce an acrosome reaction (AR) that enabled sperm to penetrate eggs. Treated sperm were incubated with liposomes for 7 min prior to insemination of zona-free hamster eggs in vitro. Sperm and eggs were incubated 3 hr at 39°C prior to fixation, staining, and examination for sperm penetration and nuclear decondensation. The percentage of motile sperm immediately after thawing as well as after treatment with liposomes had a low correlation with sire fertility (r = .39 and ≤.63, respectively). The percentage of sperm exhibiting an AR was more highly correlated with fertility (r ≤ -.85). Similar correlations were found between fertility and the penetration rates of zona-free hamster eggs or the total number of penetrating sperm. When data for two high and for two lower fertility buils were each grouped to increase information per data point the correlation between the PC12 concentration giving the maximum proportion of eggs penetrated and fertility was r = .92 (P ≤ .05). The correlation between the PC12 concentration producing the most total sperm penetrating the eggs and fertility r = .97 (P ≤ .05). It was concluded that PC12 liposomes induced an AR in bull sperm frozen-thawed in egg yolk extender. Frozen-thawed sperm from low fertility bulls require less PC12 to induce the AR and to penetrate zona-free hamster eggs than do sperm from higher fertility bulls. These differences in lipid requirements may help to provide a quick, direct laboratory assay method to estimate the fertility of frozen bull semen.
    Additional Material: 1 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/mrd.1120160206
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  • 4
    Electronic Resource
    Electronic Resource
    Characterization of a spectrophotometric technique for the estimation of fowl and turkey sperm motility (1985)
    New York, NY : Wiley-Blackwell
    Gamete Research 11 (1985), S. 169-178 
    Details
    ISSN: 0148-7280
    Keywords: spermatozoa ; motility ; fowl ; turkey ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: An objective method of estimating the motility of fowl and turkey spermatozoa, depending on their rheotactic and light-scattering properties, has been developed. From a 1- to 2-minute recorder trace of the optical density of diluted semen before and after stopping its passage through a flow cuvette, three independent constants may be simply and graphically determined. These are: ODm, the maximum optical density of semen flowing through the cuvette, shown to be dependent on the concentration of spermatozoa; % (ΔOD)m, the maximal change of optical density following cessation of flow, which has been correlated with the percentage of motile spermatozoa in the sample; and t½, the time taken for the change of optical density to reach % (ΔOD)m. This latter parameter has been correlated with forward motility of both fowl and turkey spermatozoa.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/mrd.1120110207
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  • 5
    Electronic Resource
    Electronic Resource
    Homospermic versus heterospermic insemination of zona-free hamster eggs to assess fertility of fluorochrome-labeled acrosome-reacted bull spermatozoa (1987)
    New York, NY : Wiley-Blackwell
    Gamete Research 17 (1987), S. 343-354 
    Details
    ISSN: 0148-7280
    Keywords: semen quality ; liposomes ; fluorescent stains ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Fresh spermatozoa from six bulls, with fertility ranging from 64% to 78%, (based upon 59-day nonreturn rates for 159,448 cows inseminated) were mixed with zona-free hamster eggs in 15 heterospermic pair inseminations. Five of the bulls were used in homospermic insemination studies. Prior to incubation, spermatozoa from each bull were labeled with contrasting fluorescent stains pretested for effects on spermatozoa. Equal numbers of spermatozoa were mixed and treated with liposomes of dilauroylphosphatidylcholine to induce the acrosome reaction. Spermatozoa from split ejaculates within a male competed against each other equally in the hamster egg test, indicating that the staining procedure did not affect egg penetration rates. Bulls differed in their egg penetration rates when their sperm were inseminated either homospermically or heterospermically, but the differences in the homospermic inseminations were not significantly correlated with sire fertility. The number and percentage of sperm which penetrated eggs, and the number of eggs penetrated in the heterospermic competitive tests were highly correlated with fertility (r ≥ 0.86). Therefore, egg penetration rates from heterospermic inseminations appear to be valuable indicators of fertility and much more sensitive predictors than results from homospermic inseminations.
    Additional Material: 5 Tab.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/mrd.1120170407
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