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  • 1
    Electronic Resource
    Electronic Resource
    Cytoskeletal Assemblies of Mammalian Spermatozoa (1987)
    Oxford, UK : Blackwell Publishing Ltd
    Annals of the New York Academy of Sciences 513 (1987), S. 0 
    Details
    ISSN: 1749-6632
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1749-6632.1987.tb25012.x
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  • 2
    Electronic Resource
    Electronic Resource
    FISH Analysis of Six Chromosomes in Unfertilized Human Oocytes After Polar Body Removal (2000)
    Springer
    Journal of assisted reproduction and genetics 17 (2000), S. 276-283 
    Details
    ISSN: 1573-7330
    Keywords: aneuploidy ; FISH ; oocyte ; polar body
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Purpose: To develop an improved technique for estimatingchromosomal abnormalities in human oocytes byfluorescence in situ hybridization (FISH) and to correlate theposition of single chromatids with the chromosomal status ofthe oocytes. Methods: Oocytes that were at metaphase II about17–20 hr after insemination or intracytoplasmic sperm injection(ICSI) were treated with pronase to remove the zonapellucida and polar body (PB) and then spread on slides usingHCl and Tween 20. Two rounds of FISH were performedusing direct-labeled probes: chromosomes 1, 13, 21 (round1); chromosomes X, 7, 18 (round 2). Results: Of the 63 oocytes from 18 patients (mean age,32 years), 48 (76%) had one DNA complement as expected, 9(14%) had 2 DNA complements, 3 (5%) gave incomplete FISHsignals, and 3 (5%) were not analyzable. Of the 48 oocyteswith one set of DNA, 48% were haploid, 44% were aneuploidfor one or more chromosomes, and 8% were polyploid. Wealso found an increased frequency of predivision of chromatidbivalents in aneuploid oocytes, especially for chromosome 21. Conclusions: This technique enables simultaneousassessment of six chromosomes in human oocytes, and thereforecan be useful for accurately determining the incidence andcauses of genetic imbalances in human oocytes andapparently low fertilization rates.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1009462400708
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  • 3
    Electronic Resource
    Electronic Resource
    What Is the Best Strategy for Using Embryos and Presenting Results in Assisted Reproductive Technology? A Strategy for Valid Comparisons of Results (1999)
    Springer
    Journal of assisted reproduction and genetics 16 (1999), S. 471-473 
    Details
    ISSN: 1573-7330
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1020594815171
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  • 4
    Electronic Resource
    Electronic Resource
    Localization of actin in mammalian spermatozoa: A comparison of eight species (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 216 (1986), S. 504-515 
    Details
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The distribution of monomeric and polymeric actin in spermatozoa from the bull, boar, rabbit, human, rat, mouse, golden hamster, and guinea pig has been examined by using a monoclonal antiactin antibody and NBD-phallacidin. Actin was present in sperm from each species. When the monoclonal antibody was used, there was a species-specific distribution and intensity of fluorescence, but no generalized pattern. Specific fluorescence was noted in the neck and principal piece of human sperm; in the postacrosomal region, neck, and midpiece of bull and boar sperm; in the postacrosomal region, neck, and principal/equatorial segment border of rabbit sperm; in the neck region of hamster sperm; and in the neck, midpiece, and principal piece of rat, mouse, and guinea pig sperm. Sperm from all eight species displayed no specific fluorescence with NBD-phallacidin, indicating that actin was present in a nonfliamentous form. SDS extracts of sperm were analyzed by SDS-PAGE and Western blotting; in sperm from each species, a 42-kD protein with specific affinity for the monoclonal antibody was present.
    Additional Material: 20 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/ar.1092160407
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  • 5
    Electronic Resource
    Electronic Resource
    Localization of actin in human, bull, rabbit, and hamster sperm by immunoelectron microscopy (1988)
    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 221 (1988), S. 599-610 
    Details
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Mammalian spermatozoa have previously been shown to contain actin, but its subcellular localization and function have not been elucidated. In this study, actin has been localized at the ultrastructural level in human, bull, rabbit, and golden hamster spermatozoa by a monoclonal antiactin antibody and a preembedding immunogold labeling technique. Specific labeling was localized 1) around the connecting piece in the neck region of sperm from all four species, although a species-specific pattern was evident; 2) on the external surface of the fibrous sheath of human sperm; 3) in the perinuclear space underlying the postacrosomal sheath of bull and rabbit sperm; and 4) between the plasma membrane and outer acrosomal membrane along the concave margin of the hamster sperm head. SDS-PAGE and Western blots immunostained with the monoclonal antibody confirmed the presence of actin in SDS extracts of Percoll-purified sperm from each species.
    Additional Material: 23 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/ar.1092210206
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  • 6
    Electronic Resource
    Electronic Resource
    Further ultrastructural observations on the sperm head of the plains mouse, Pseudomys australis (Rodentia: Muridae) (1987)
    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 217 (1987), S. 240-249 
    Details
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Ultrastructural and histochemical techniques have been utilized to study selected aspects of the fine structure of the three-hooked sperm head of the plains mouse, Pseudomys australis. The peripheral layer of the two ventral hooks was found to consist of a continuation of the postacrosomal dense lamina. Parallel ridges connected the dense lamina and plasma membrane in the postacrosomal region and ventral hooks. Both these regions stained intensely with silver nitrate. The distribution of action filaments in the hooks was investigated using NBD-phallacidin. Fluorescence was more intense in the apical regions of the ventral hooks, and two bands of fluorescence extended caudally from their base. It was also shown that the equatorial segment of the acrosome extended onto the dorsal hook. The structural features of the three hooks are discussed in relation to their possible functional significance.
    Additional Material: 19 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/ar.1092170304
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  • 7
    Electronic Resource
    Electronic Resource
    Membrane domains in guinea pig sperm and their role in the membrane fusion events of the acrosome reaction (1988)
    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 220 (1988), S. 267-280 
    Details
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: In this study, we have examined the structure of domains of the periacrosomal plasma membrane (PM) and outer acrosomal membrane (OAM) of guinea pig sperm and defined their fate during the membrane fusion events of the acrosome reaction. Cauda epididymal sperm were arranged in rouleaux, joined by periacrosomal PM “junctional” zones; in these zones, the PMs were linked by cross bridges formed from a paracrystalline glycocalyx. Bridging elements linked the PM to the OAM on the ventral (concave) but not dorsal (convex) aspect of the apical segment. Parallel filaments were associated with the luminal face of the OAM overlying the dorsal surface of the apical segment. Sperm were induced to undergo a “synchronous” acrosome reaction after preincubation in Ca2+-free medium containing lysolecithin, by the addition of Ca2+. Fusion between the OAM and PM occurred at the boundaries but not within the PM “junctional” zones over the apical segment. In nonjunctional regions on the dorsal surface of the apical segment, sheets of unfenestrated hybrid membranes and parallel arrays of hybrid membrane tubules formed, while branching arrays of hybrid membrane tubules and vesicles were observed on the ventral surface. In the principal segment, networks of branching hybrid membrane tubules initially formed but later transformed into vesicles. Hence, the lysolecithin-mediated guinea pig sperm acrosome reaction involves a complex sequence of membrane fusions, which differs in domains of the periacrosomal PM and OAM. Stable nonfusigenic domains are present in both the PM and OAM of the apical segment; membrane-associated assemblies may maintain these domains and may also provide direction to some of the membrane fusion events of the acrosome reaction.
    Additional Material: 23 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/ar.1092200307
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  • 8
    Electronic Resource
    Electronic Resource
    Ultrastructural analysis of the acrosome reaction in a population of single guinea pig sperm (1991)
    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 229 (1991), S. 186-194 
    Details
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Cauda epididymal guinea pig spermatozoa are arranged in rouleaux, with the sperm heads stacked one on top of the other; the plasma membranes over the apical segment of the acrosomes of adjacent sperm are linked and form non-fusigenic “junctional” zones. A complex structural and temporal sequence of membrane fusions occurs during the acrosome reaction of guinea pig sperm in rouleaux. In this study, we have devised a procedure for dispersing the rouleaux and isolating a population of single, motile guinea pig sperm, and have investigated the ultrastructural features of the acrosome reaction in single sperm to determine if the pattern of membrane fusions is different from sperm in rouleaux. The rouleaux were dispersed using trypsin, and damaged cells were removed by passing the sperm suspension through a glass bead column; a population of 70-90% motile, acrosome-intact, single sperm was obtained. Sperm were then induced to undergo lysolecithin-mediated, “synchronous” acrosome reactions, and processed for transmission electron microscopy. The acrosome reaction involved a complex sequence of membrane fusions between the plasma membrane (PM) and outer acrosomal membrane (OAM). On the convex surface of the apical segment, sheets of hybrid membrane and parallel arrays of hybrid membrane tubules formed; filaments were associated with the luminal surface of the residual OAM in these regions. Hybrid membrane vesicles were produced on the concave surface of the apical segment, but fusion was delayed relative to the convex surface. In the principal segment, branching arrays of hybrid membrane tubules formed and later vesiculated. Hence, in single guinea pig sperm, the sequence of membrane fusions is similar to sperm in rouleaux except that fusion occurs in regions of the apical segment which form the non-fusigenic PM “junctional” zones in rouleaux. The results suggest that, regardless of whether the acrosome reaction in vivo occurs before or after rouleaux dispersion, it will involve a complex sequence of membrane fusions which is determined by the structural properties of the OAM and PM.
    Additional Material: 13 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/ar.1092290205
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  • 9
    Electronic Resource
    Electronic Resource
    Detection of chromosome 17- and X-bearing human spermatozoa using fluorescence in situ hybridization (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 33 (1992), S. 189-194 
    Details
    ISSN: 1040-452X
    Keywords: Spermatozoa ; Diploid ; Disomy ; Fluorescence in situ hybridization ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Fluorescence in situ hybridization (FISH) with DNA probes specific to chromosomes 17 and the X has been applied to human ejaculated sperm. After sperm nuclei were decondensed with EDTA and DTT, biotinylated alpha satellite DNA probes TR17 and TRX were separately used on preparations from thirteen healthy donors. After hybridization 96% of sperm were labelled with the TR17 probe and 48% of sperm were labelled with the TRX probe. Frequencies of 0.33% disomic 17 and 0.29% disomic X sperm were found. The frequencies of diploid sperm were assessed as 0.37% using the TR17 probe and 0.20% using the TRX probe which labelled only one half of the sperm; after correcting the result from the X-probe to 0.40% the two frequencies are very similar. © 1992 Wiley-Liss, Inc.
    Additional Material: 10 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/mrd.1080330211
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  • 10
    Electronic Resource
    Electronic Resource
    Simultaneous detection of X- and Y-bearing human sperm by double fluorescence in situ hybridization (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 34 (1993), S. 308-313 
    Details
    ISSN: 1040-452X
    Keywords: Sex chromosomes ; Human sperm ; Haploid ; Diploid ; Double fluorescence in situ hybridization ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Double fluorescence in situ hybridization (FISH) was used to detect sex chromosomes in decondensed human sperm nuclei. Biotinylated X chromosome specific (TRX) and digoxigenin-labeled Y chromosome specific (HRY) probes were simultaneously hybridized to sperm preparations from 12 normal healthy donors. After the hybridization, the probes were detected immuno-cytochemically, using two different and independent affinity systems. Ninety-six percent of the 12,636 sperm showed fluorescent labeling, of which 47.4% were haploid X and 46.8% were haploid Y. A frequency of 0.46% of XX-bearing sperm (0.28% disomic, 0.18% diploid) and 0.38% YY-bearing sperm (0.21% disomic, 0.17% diploid) was found. The overall proportions of X- and Y-bearing sperm in the ejaculates were 47.9% and 47.2%, respectively, which was not significantly different from the expected 50:50 ratio. In addition 0.21% of cells appeared to be haploid XY-bearing sperm, 0.62% were diploid XY-bearing cells, and 0.05% of cells were considered to be tetraploid cells. The application of double FISH to human sperm using X-chro-mosome and Y-chromosome probes has allowed a more accurate assessment of the sex chromosal complements in sperm than single FISH method and quinacrine staining for Y-bodies. © 1993 Wiley-Liss, Inc.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/mrd.1080340311
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