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  • Articles  (30)
  • sperm  (30)
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  • American Association for the Advancement of Science
  • 1980-1984  (30)
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  • Wiley-Blackwell  (30)
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  • Biology  (30)
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  • 1
    Electronic Resource
    Electronic Resource
    The interaction of cAMP with modeled bull sperm (1983)
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 3 (1983), S. 199-210 
    Details
    ISSN: 0886-1544
    Keywords: sperm ; flagellum ; motility ; cAMP ; freeze-thawing ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Demembranated and membrane disrupted bull sperm models exhibit an increase in motility when exposed to cAMP. Tritium-labeled cAMP was used to locate the initial site of action of cAMP in the modeled sperm preparations. cAMP did not bind selectively to the modeled cells, and the presence or absence of plasma membrane fragments on the models did not significantly alter this result. When suspension medium taken from modeled sperm preparations was subjected to gel filtration on Sephadex G25-150 columns, cAMP bound to a high molecular weight component that eluted with the void volume. The responsible binding factor is a soluble component that is released when the plasma membranes of the sperm are disrupted during the modeling procedure. To test the importance of the cAMP binding factor, modeled sperm were centrifuged, the super-natant solution was decanted, and the cells were resuspended in fresh medium. After this treat-ment the cells could be restored to motility with Mg-ATP but no longer exhibited a response to cAMP. Furthermore, addition of cAMP binding factor isolated by gel filtration partially restored the response of these sperm to cAMP. Investigation of the properties of the cAMP-binding factor have confirmed that it is specific for cAMP, with a much lower affinity for AMP and cGMP. In the pre-sence of a large excess of unlabeled cAMP the labeled complex has a half-life of approximately 1 hour. Our results indicate that the action of cAMP on the motility of modeled sperm is mediated by its attachment to a high molecular weight, soluble component of the cell cytoplasm.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/cm.970030208
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  • 2
    Electronic Resource
    Electronic Resource
    Quantitation of avian spermatozoan motility: Neurochemical regulation (1980)
    New York, NY : Wiley-Blackwell
    Gamete Research 3 (1980), S. 17-24 
    Details
    ISSN: 0148-7280
    Keywords: sperm ; motility ; neurochemical ; paraoxon ; acetylcholine ; cholinesterase ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The hypothesis that motility of avian sperm is regulated by acetylcholine was examined by treating rooster (Gallus domesticus) sperm with choline analogs and paraoxon, an inhibitor of colonesterases. Acetylcholine chloride (AChCl) was most effective, acetylthiocholine iodide and butyrylthiocholine iodide were less effective, and choline chloride was ineffective in stimulating sperm motility. Histochemical localization of cholinesterase activity with the electron microscope showed enzyme activity to be associated with membranes of the head and within fibrillar components of the tail. Increasing concentrations of paraoxon decreased cholinesterase activity and increased sperm motility. The data provide evidence that the motility of avian sperm, like that of mammal and sea urchins, may be regulated in part by a system with similarities to the cholinergic neurotransmitter system.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/mrd.1120030104
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  • 3
    Electronic Resource
    Electronic Resource
    Artificial fertilization of gametes from the South African clawed frog, Xenopus laevis (1980)
    New York, NY : Wiley-Blackwell
    Gamete Research 3 (1980), S. 45-57 
    Details
    ISSN: 0148-7280
    Keywords: clawed frog ; egg ; fertilization ; jelly coat ; motility ; sperm ; Xenopus laevis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A reproducible and effective method for fertilization eggs of Xenopus laevis was developed based of systematic manipulation of environmental factors. The effects of varying concentrations of individual components of a fertilization medium were tested by measuring jelly swelling, sperm motility, and sperm longevity. Results were used to develop an improved medium for fertilization, consisting of 41.25 mM NaCl, 1.25 mM KCl, 0.25 mM CaCl2, 0.0625 mM MgCl2, 0.5 mM Na2HPO4, 2.5 mM HEPES, 1.9 mM NaOH, final pH(2°) 7.8.
    Additional Material: 12 Ill.
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    URL: http://dx.doi.org/10.1002/mrd.1120030106
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  • 4
    Electronic Resource
    Electronic Resource
    Fragmentation of condensed chromatin in nuclei of a plant sperm, pteridium aquilinum (L.) Kuhn (1980)
    New York, NY : Wiley-Blackwell
    Gamete Research 3 (1980), S. 291-298 
    Details
    ISSN: 0148-7280
    Keywords: condensed chromatin ; sperm ; Pteridium ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Nuclei of pteridium sperm have been dispersed by turbulence in natural or slightly alkaline buffer after stripping off the cytoplasm with nonionic detergent. The nuclei tended to break up into fragments arranged in a linear order. These fragments fluoresced brightly with acridine orange as did intact nuclei. Grounds are given for identifying the smaller fragments with chromosomes. It is proposed that the sperm nucleus of British Pteridium, possibly an autotetrapolid, consists of a sequence of paired homologues.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/mrd.1120030311
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  • 5
    Electronic Resource
    Electronic Resource
    Evidence for nonmuscle myosin in bovine ejaculated spermatozoa (1981)
    New York, NY : Wiley-Blackwell
    Gamete Research 4 (1981), S. 499-506 
    Details
    ISSN: 0148-7280
    Keywords: sperm ; nonmuscle myosin ; affinity column ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Mammalian spermatozoa contain nonmuscle actin and many of the components of regulatory systems thought to be involved in nonmuscle actin-myosin function. An actin-stimulated adenosine triphosphate hydrolase (ATPase) activity has been fractionated from bovine ejaculated spermatozoa by immobilized-actin affinity chromatography. The actinstimulated ATPase activity has a specific activity (0.04 ± 0.02 mM phosphate released/min/mg protein) similar to nonmuscle myosins from other mammalian cells and tissues, but it does not have appreciable K+-EDTA ATPase activity. The sperm actin-myosin may function in sperm morphogenesis in the seminiferous tubule, in capacitated spermatozoa undergoing an acrosome reaction, or in decondensation of the sperm nucleus after fertilization.
    Additional Material: 1 Ill.
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    URL: http://dx.doi.org/10.1002/mrd.1120040603
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  • 6
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    Stimulation of hamster sperm capacitation and acrosome reaction in vitro by glucose and lactate and inhibition by the glycolytic inhibitor α-chlorohydrin (1981)
    New York, NY : Wiley-Blackwell
    Gamete Research 4 (1981), S. 515-523 
    Details
    ISSN: 0148-7280
    Keywords: glucose ; glycolysis ; lactate ; sperm ; capacitation ; acrosome reactions ; α-chlorohydrin ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Studies were made of the effects of D(+)-glucose, L-lactate and pyruvate on in vitro capacitation and acrosome reactions (AR) of hamster sperm using a more “defined” medium that that used in previous similar studies. In the absence of glucose or lactate, sperm underwent very few AR and activation (whiplash-like motility characteristic of capacitated hamster sperm) was reduced compared to those events in sperm preincubated in the presence of glucose plus lactate plus pyruvate. Glucose and pyruvate supported more AR than glucose alone, but less than glucose, lactate, and pyruvate. The glycolytic inhibitor α-chlorohydrin (10 μm) inhibited AR by 50% and reduced activation by less. When glucose was added to sperm incubated 2 hr with pyruvate and lactate, the number of AR observed after 4 hr was the same as that obtained when glucose was present throughout the incubation. When glucose was added after 3.5 hr, AR were delayed for 1 hr and lower numbers of sperm underwent AR. In the presence of lactate and pyruvate, 0.38 mM glucose was able to support activation and AR as well as 3.24 mM glucose. These results indicate that exogenous glucose and lactate are necessary for in vitro capacitation and AR of hamster sperm; only low levels of exogenous glucose are required; exogenous glucose is not required during the first 2 hr of capacitation; and glycolytic activity is necessary for capacitation and the AR.
    Additional Material: 1 Ill.
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    URL: http://dx.doi.org/10.1002/mrd.1120040605
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  • 7
    Electronic Resource
    Electronic Resource
    Architecture of the hamster oocyte-cumulus complex (1984)
    New York, NY : Wiley-Blackwell
    Gamete Research 9 (1984), S. 261-272 
    Details
    ISSN: 0148-7280
    Keywords: fertilization ; cumulus ; oocyte ; sperm ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The structure of the hamster oocyte-cumulus complex (OCC) has been analyzed to help understand how mammalian sperm penetrate the investing coats of the oocyte. At ovulation, oocytes of most mammalian species are surrounded by a zona pellucida, corona radiata, and cumulus layer. Cells of the cumulus and corona radiata are separated by an extracellular matrix (ECM) that contains hyaluronic acid. In hamsters, the diameter of mature follicular OCCs is 0.61 ± 0.12 mm, whereas in freshly ovulated OCCs in culture medium it is 0.78 ± 0.15 mm. This indicates the OCC expands at or after ovulation. The corona radiata is 1-4 cell layers deep, and corona radiata cells are closely packed (center-to-center distances between adjacent cells in follicular OCCs averaged 14 ± 3 μm). The cumulus layer is 5-8 cell layers deep, and intercellular spaces are much larger (center-to-center distances between adjacent cumulus cells in follicular OCCs averaged 50 ± 20 μm). An ovulated OCC has an approximate volume of 248 × 106 μm3 and was estimated to contain approximately 5,700 cells. Studies with stretched OCCs show that ink particles can readily penetrate the extracellular spaces of the cumulus and corona radiata layers and interact with the ECM, staining it black. At the periphery of the OCC, the ECM appeared discontinuous and formed “cords” containing cumulus cells, whereas closer to the corona radiata the matrix completely filled intercellular spaces. Ink penetrated the corona radiata, but the ECM was difficult to visualize because of the close packing of cells in this layer. Our observations on unfixed OCCs show that cell packing becomes more dense proceeding from the periphery of the OCC to the zona pellucida and that the ECM at the periphery differs from the matrix deeper in the OCC. These data suggest that an incoming sperm would find penetration of the investing coats to increase in difficulty (that is physical resistance would increase) as it got closer to the oocyte. We have also examined the effects of processing for microscopy on the structure of the OCC. Both standard fixation procedures and fixation in the presence of ruthenium red caused condensation of the ECM, especially in the cumulus layer, and thus produced a significant decrease in the diameter of the OCC.
    Additional Material: 7 Ill.
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    URL: http://dx.doi.org/10.1002/mrd.1120090304
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  • 8
    Electronic Resource
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    Normal levels of zinc and sulfhydryls in morphologically abnormal populations of spermatozoa from moderately zinc-deficient rats (1984)
    New York, NY : Wiley-Blackwell
    Gamete Research 9 (1984), S. 375-386 
    Details
    ISSN: 0148-7280
    Keywords: rat ; zinc-deficient ; sperm ; dense fibers ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Zinc is required for spermatogenesis in mammals and is concentrated in the dense outer fibers of the sperm tail, where it is associated with cysteine-rich protein. To investigate the effects of marginal zinc deficiency upon dense fiber formation and upon sperm quality in general, weanling Sprague-Dawley rats were administered a commercial low-zinc diet, supplemented with phytate, for approximately 60 days, and were compared with controls fed the same diet plus 50 ppm zinc in their drinking water. The following characteristics of the zinc-deficient rats were significantly lower than in the controls: body weight, testis weight, epididymis weight, seminal vesicle weight, sperm content of the cauda epididy-midis, sperm motility, testis zinc, and hair zinc. By contrast, the levels of sperm zinc and sperm sulfhydryls were the same in the zinc-deficient and control rats. The zinc-deficient rats displayed a highly variable spectrum of sperm defects, which included decapitation, disorganized and redundant tail elements, and superfluous cytoplasm. However, abortive dense fiber development was only rarely observed. Apparently, even when availability of zinc is limited and reduced sperm production ensues, elaboration of dense fibers rich in zinc and sulfhydryls continues to be obligatory for the completion of spermiogenesis.
    Additional Material: 4 Ill.
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    URL: http://dx.doi.org/10.1002/mrd.1120090403
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  • 9
    Electronic Resource
    Electronic Resource
    Sperm head structure in the hydromyinae (rodentia:Muridae): A further evolutionary development of the subacrosomal space in mammals (1984)
    New York, NY : Wiley-Blackwell
    Gamete Research 10 (1984), S. 31-44 
    Details
    ISSN: 0148-7280
    Keywords: subacrosomal space ; sperm ; murid rodents ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: This scanning and transmission electron microscopical study has been performed to determine the extent of the development of the subacrosomal space in the sperm head of various members of the subfamily, the Hydromyinae, which is a diverse group of murid rodents occurring in various habitats in Australia. Sperm of all species from the three tribes of Hydromyini, Uromyini, and Conilurini investigated in this study had a head with three hooks although their absolute and relative lengths varied markedly. The top hook was similar in structure to that present in many other murid rodents and had a typical pseudoperforatorium present. The two ventral hooks had nuclear material basally, apical to which a large extension of the subacrosomal space occurred. Studies of testicular sections showed that the ventral hooks were formed late in spermiogenesis largely after condensation of the nucleus during which time electron dense material accumulated within them. No structure homologous to these hooks has been found in sperm from any other mammalian group, but, as the three tribes of Hydromyinae probably diverged about 20 million years ago, the extension of the subacrosomal space presumably evolved prior to this date.
    Additional Material: 41 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/mrd.1120100105
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  • 10
    Electronic Resource
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    An improved method to visualize human sperm chromosomes using zona-free hamster eggs (1984)
    New York, NY : Wiley-Blackwell
    Gamete Research 10 (1984), S. 233-239 
    Details
    ISSN: 0148-7280
    Keywords: chromosome ; spermatozoa ; sperm ; egg ; zona-free ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: An improved method for human sperm chromosome preparation is described. Improvements include (1) the use of a sperm population with high motility and normal morphology for insemination, (2) the insemination and postinsemination culture of zona-free eggs in Holmes medium, which does not require serum supplementation, (3) control of sperm concentration at insemination to avoid heavy polyspermy, (4) reduction of the occurrence of egg agglutination by placing the medium for postinsemination culture in a ring or crescent shape instead of a droplet, and (5) application of a two-step fixation method to augment the efficiency of chromosome preparation.
    Additional Material: 1 Ill.
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    URL: http://dx.doi.org/10.1002/mrd.1120100303
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  • 11
    Electronic Resource
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    Influence of the concentration of sperm on the percentage of eggs fertilized for three strains of mice (1984)
    New York, NY : Wiley-Blackwell
    Gamete Research 10 (1984), S. 415-422 
    Details
    ISSN: 0148-7280
    Keywords: mouse ; fertility ; fertilization ; sperm ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: This study was conducted to determine the optimal concentration of sperm to use for the insemination of females to detect differences among strains of mice in the percentage of eggs fertilized. Female ICR mice were inseminated with sperm of concentrations ranging from 0.25 to 8 × 106/50 μl from males of either DBA/2N, CF1, or C57BL/6N strains. Differences among strains were detected only when approximately 50% of the eggs were fertilized but not when each of the strains fertilized either a high or low percentage of eggs. The optimal concentration of sperm therefore was the concentration that gave approximately 50% fertilized eggs.
    Additional Material: 1 Ill.
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    URL: http://dx.doi.org/10.1002/mrd.1120100407
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  • 12
    Electronic Resource
    Electronic Resource
    Rabbit testis proacrosin: Immunological similarities between testis, sperm proacrosins and the initially formed testis acrosin (1980)
    New York, NY : Wiley-Blackwell
    Gamete Research 3 (1980), S. 241-246 
    Details
    ISSN: 0148-7280
    Keywords: testis ; sperm ; proacrosin ; acrosin ; immunological ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Anti-rabbit proacrosin IgG was prepared from goat serum following immunization with a homogeneous preparation of rabbit testis proacrosin. The “auto-activation” products of purified testis proacrosin were separated into 68,000 and 34,000 molecular weight (mol wt) acrosins by Sephadex G-100 column chromatography. Immunodiffusion analysis of testis and epididymal sperm proacrosins and acrosins on agarose gel against goat anti-rabbit testis proacrosin showed immunological identity between rabbit testis and sperm proacrosins and the initial testis acrosin (mol wt 68,000). However, the 34,000 mol wt form of testis acrosin showed weaker reaction with the antibody and only partial identity with the proacrosin and the 68,000 mol wt form of acrosin. These results suggest that there is no major structural difference between testis and sperm proacrosins and between proacrosin and the 68,000 mol wt acrosin, but such a structual change occurs when the 34,000 mol wt acrosin is formed.
    Additional Material: 1 Ill.
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    URL: http://dx.doi.org/10.1002/mrd.1120030306
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  • 13
    Electronic Resource
    Electronic Resource
    Optimization of culture conditions for the formation of sperm cells in pollen tubes of tradescantia (1981)
    New York, NY : Wiley-Blackwell
    Gamete Research 4 (1981), S. 35-40 
    Details
    ISSN: 0148-7280
    Keywords: sperm ; pollen tube ; culture media ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A culture medium and culture conditions are described that enable generative cell division and sperm formation to occur in a large proportion (greater than 70%) of the pollen tubes of Tradescantia paludosa within six to eight hours of culture of pollen. The nature of the nitrogen source, speed of shaking, and ratio of pollen to medium are important parameters in determining the extent of sperm formation. Addition of the plant hormones indole acetic acid, gibberellic acid, and kinetin to the growth medium does not influence generative cell division.
    Additional Material: 4 Ill.
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    URL: http://dx.doi.org/10.1002/mrd.1120040106
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  • 14
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    Chromatin transitions in the fertilizing sperm nucleus of the sea urchin, strongylocentrotus purpuratus (1982)
    New York, NY : Wiley-Blackwell
    Gamete Research 5 (1982), S. 181-190 
    Details
    ISSN: 0148-7280
    Keywords: fertilization ; sperm ; chromatin ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Electron microscopic analysis of fertilization in the sea urchin, Strongylocentrotus purpuratus, has been carried out in an effort to establish the sequence of events involving dispersion of the paternal chromatin. Subsequent to loss of the nuclear envelope the condensed sperm chromatin begins to disperse under the influence of egg cytoplasmic factors. However, this process does not proceed at a uniform rate as is observed in other species examined to date. Portions of the paternal genome rapidly transform into dispersed chromatin while other adjacent regions disperse at a reduced rate. This variation in the time sequence of dissociation of the paternally derived chromosomes results in a reticulum of electron lucent and electron dense chromatin within the developing male pronucleus. As the paternally derived chromatin is dispersing and migrating centrad, membranous vesicles of maternal origin become aligned along the peripheral aspect of the chromatin. Deposition of a continuous bilaminar nuclear envelope around the dispersing sperm chromatin results in the formation of the definitive male pronucleus. At the time the male pronucleus is formed the paternally derived chromosomes have not completely dispersed and are visualized as a reticulum of condensed and dispersed chromatin. These results indicate that not all the paternally derived chromatin is modified in the same manner during pronuclear development.
    Additional Material: 5 Ill.
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    URL: http://dx.doi.org/10.1002/mrd.1120050208
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  • 15
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    Evidence for acrosin-like enzyme in sperm extract and its involvement in fertilization of the ascidian, halocynthia roretzi (1982)
    New York, NY : Wiley-Blackwell
    Gamete Research 5 (1982), S. 291-301 
    Details
    ISSN: 0148-7280
    Keywords: ascidian ; sperm ; acrosin-like enzyme ; protease ; lysin ; fertilization ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The presence of a protease has been demonstrated in sperm of the solitary ascidian, Halocynthia roretzi, by using t-butyloxycarbonyl-L-Val-L-Pro-L-Arg-4-methylcoumaryl-7-amide (Boc-Val-Pro-Arg-MCA) and other arginyl or lysyl MCA derivatives as substrates. Several properties of the enzyme were investigated in a crude extract. The activity had a pH optimum near 8.0 and was enhanced by the addition of CaCl2. The Km value of 87μM was determined for Boc-Val-Pro-Arg-MCA under the optimal conditions. An apparent molecular weight was estimated to be 35,000 by gel filtration. The enzyme was inhibited with diisopropyl fluorophosphate, leupeptin, antipain, p-aminobenzamidine, Val-Pro-Arg-CH2Cl, and soybean trypsin inhibitor, but scarcely inhibited with chymostatin, elastatinal, p-chloromercuribenzoic acid, tosyl-Lys-CH2Cl, and tosyl-Phe-CH2Cl. Boc-Val-Pro-Arg-MCA, the most susceptible of the substrates examined, showed the most effective inhibition against fertilization of ascidian eggs.Thus, this enzyme in ascidian sperm extract has features closely similar to mammalian acrosin [EC 3.4.21.10], and we conclude that the enzyme is involved in fertilization as one of the lysins.
    Additional Material: 2 Ill.
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    URL: http://dx.doi.org/10.1002/mrd.1120050309
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  • 16
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    A modified column for sperm isoelectric focusing (1983)
    New York, NY : Wiley-Blackwell
    Gamete Research 7 (1983), S. 325-329 
    Details
    ISSN: 0148-7280
    Keywords: isoelectric focusing apparatus ; sperm ; protein ; density gradient ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A modified U-shaped column is described for efficient isoelectric focusing of spermatozoa, other cells, and protein. The washed spermatozoa of the rabbit showed a PI of 4.4. After sonication, heads and tails focused at same pH, indicating similar and equal charge.
    Additional Material: 6 Ill.
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    URL: http://dx.doi.org/10.1002/mrd.1120070404
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  • 17
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    Plasma membrane structure in spermatogenic cells of the swordtail (teleostei, Xiphophorus helleri) (1983)
    New York, NY : Wiley-Blackwell
    Gamete Research 7 (1983), S. 309-324 
    Details
    ISSN: 0148-7280
    Keywords: Teleost ; sperm ; membrane ; ultrastructure ; lectin-binding ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The plasma membrane of spermatogenic cells of the teleost Xiphophorus helleri was examined ultrastructurally and cytochemically in order to characterize the temporal development of the membrane specializations characteristic of the mature spermatozoon. Mature sperm display a mosaic distribution of Concanavalin A and Ricinus comrnunis I binding sites; the anterior region of the head displays an intense binding that is not seen in other surface regions. This asymmetric binding is evident in early spermatids and the area of lectin binding appears associated with the plasma membrane overlying the nucleus. Transmission electron microscopy reveals that the plasma membrane over the anterior region of the head is characterized by an ordered glycocalyx and a tight adherence to the underlying nucleus. Additional membrane differentiations were revealed both in the midpiece region where a “submitochondrial net” is attached to the plasma membrane and at the base of the axoneme where the plasma membrane possesses a “collar-like” arrangement of circumferential rings. The possible functions of these differentiations, as well as their correlation to differentiations seen in sperm of other animal groups, are discussed.
    Additional Material: 15 Ill.
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    URL: http://dx.doi.org/10.1002/mrd.1120070403
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  • 18
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    Possible involvement of a sialylated component of the sperm plasma membrane in sperm-zona interaction in the mouse (1984)
    New York, NY : Wiley-Blackwell
    Gamete Research 10 (1984), S. 153-163 
    Details
    ISSN: 0148-7280
    Keywords: mouse ; sperm ; egg ; sperm plasma membrane ; in vitro ; binding ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have studied the molecular mechanisms of gamete interaction in vitro in the laboratory mouse, Mus musculus. In particular, we were interested in whether this interaction is similar to a lectin-hapten-mediated process. Inhibition of sperm-zona binding was examined using various concentrations (.25 mM to 50 mM) of different sugars (sialic acid α-methylmannose, glucose, fucose, galactose, and N-acetyl-glucosamine). Sperm-zona binding was significantly decreased when eggs were pretreated with 10 mM of sialic acid or α-methylmannose but not by other sugars tested. Furthermore, treatment of capacitated sperm with neuraminidase destroyed their ability to bind and fertilize eggs. We have also used a specific lectin for sialic acid from the horseshoe crab (Limulus polyphemus) to agglutinate mouse sperm. The lectin (.120 ng/ml) mediated agglutination of mouse sperm (105 sperm/ml) was routinely observed to increase from a 10% agglutination immediately following their isolation from the epididymis to 100% agglutination 90 minutes later. Collectively, these results suggest the appearance of specific sugar moieties on the surface of the sperm plasma membrane which, in this particular species of mouse, are sialylated glycoproteins acting as ligands for specific receptors on the surface of the egg. These are the first data to indicate that sperm-egg recognition and attachment is a lectin-hapten-mediated process in the mouse.
    Additional Material: 6 Ill.
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    URL: http://dx.doi.org/10.1002/mrd.1120100207
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  • 19
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    Rabbit spermatozoa undergo an acrosome reaction in the presence of glycosaminoglycans (1983)
    New York, NY : Wiley-Blackwell
    Gamete Research 8 (1983), S. 11-19 
    Details
    ISSN: 0148-7280
    Keywords: sperm ; acrosome reaction ; glycosaminoglycans ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Rabbit-ejaculated spermatozoa were incubated in a chemically defined medium containing comercially available glycosaminoglycans (GAGs) at concentrations ranging from 0.1 to 100 μg/ml. Sperm were stained and examined for the degree of acrosome reaction and viability after 9 h of incubation. There were significant dose and treatment effects of the induction of the acrosome reaction. Viability did not differ significantly betweendoses or treatment. Heparin enhanced the acrosome reaction between concentrations of 0.1 to 1.0 μg/ml, whereas higher levels depressed the percentage of sperm undergoing the acrosome reaction. Seminal plasma added to sperm cultures depressed the stimulatory effect of GAGs. Treatment of chondroitin-4-sulfate with chondro-4-sulfatase prohibited the stimulatory effect. It is concluded that GAGs, components of the female reproductive tract, may promote the acrosome reaction so that successful fertilization can occur.
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    URL: http://dx.doi.org/10.1002/mrd.1120080103
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    Adenylate cyclase activity of membrane fractions isolated from sea urchin sperm (1984)
    New York, NY : Wiley-Blackwell
    Gamete Research 9 (1984), S. 87-97 
    Details
    ISSN: 0148-7280
    Keywords: sperm ; membrane ; adenylate-cyclase ; cAMP ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The adenylate cyclase activity of sperm membrane fragments isolated from Lytechinus pictus sperm according to Cross [20] has been studied. Two distinct fractions preferentially coming from the flagellar plasma membrane are obtained. Surface I125-labeling experiments performed by Cross [20] indicate that these membranes are representative of the entire sperm plasma membrane. Both fractions are enriched in their adenylate cyclase activity: the specific activity of the top membranes is eightfold higher than in whole sperm, whereas that of the middle membranes is 15-fold higher. The cyclase seems to be associated with the membranes. Lytechinus pictus egg jelly has no effect or slightly inhibits the adenylate cyclase activity of the isolated sperm plasma membrane fragments. Mg++ and Na+ stimulated their cyclase activity about sevenfold at 2.5 mM Mn++ and 3.2 mM ATP. At this ATP to Mn++ ratio, high concentrations of Ca++ have a small stimulatory effect.
    Additional Material: 5 Ill.
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    URL: http://dx.doi.org/10.1002/mrd.1120090108
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    Vitelline layer lytic activity in sperm extracts of sea urchin, Hemicentrotus pulcherrimus (1981)
    New York, NY : Wiley-Blackwell
    Gamete Research 4 (1981), S. 193-202 
    Details
    ISSN: 0148-7280
    Keywords: lysin ; protease ; sperm ; sea urchin ; vitelline layer ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A factor which dissolves the vitelline layer was extracted from sperm of the sea urchin, Hemicentrotus pulcherrimus. Turbidity of the suspension was reduced when isolated vitelline layers were mixed with this sperm factor. When the mixture was subjected to SDS polyacrylamide gel electrophoresis, some of the protein bands of the vitelline layer were seen to be missing. The lytic activity of the factor was heat labile, completely inhibited by L-1-tosyl-amide-2-phenyl-ethylchloromethyl ketone and partially inhibited by soybean trypsin inhibitor. Chymotrypsin activity was detected, but not trypsin, arylsulfatase, or glycosidase. These results suggest that a chymotrypsin-like enzyme participates in lysis of the vitelline layer by the fertilizing spermatozoon.
    Additional Material: 7 Ill.
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    URL: http://dx.doi.org/10.1002/mrd.1120040304
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    22Na+ and 86Rb+ fluxes in spermatozoa and oocytes of arbacia punctulata (1982)
    New York, NY : Wiley-Blackwell
    Gamete Research 6 (1982), S. 135-143 
    Details
    ISSN: 0148-7280
    Keywords: sperm ; oocyte ; 22Na+ ; 86Rb+ ; fluxes ; Arbacia ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The fluxes of 22Na+ and 86Rb+ in Arbacia sperm and oocytes were studied in order to determine how these cells carry out cation exchange with the sea environment. The uptake of these ions by serum followed a pattern of early rapid influx (initial 0.5 min) and subsequent efflux (1-3 min) followed by a gradual uptake (after 3 min). Neither the uptake nor the efflux of these cations by Arbacia sperm were affected by ouabain, suggesting that influx and efflux of 22Na+ and 86Rb+ in Arbacia sperm occur predominantly by passive transport. The 22Na+ uptake by Arbacia oocytes showed a steady increase after an initial rapid uptake. A slight but significant inhibition of 22Na+ uptake was observed with ouabain. However, 86Rb+ uptake by the oocytes reached an early equilibrium and was not affected by ouabain. The uptake of Rb+ by Arbacia oocyte is by passive transport while that of Na+ is both by passive and active transport.
    Additional Material: 5 Ill.
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    URL: http://dx.doi.org/10.1002/mrd.1120060207
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    Fusion of hamster and pig zona-free eggs stimulated by boar and guinea pig sperm at fertilization in vitro (1982)
    New York, NY : Wiley-Blackwell
    Gamete Research 6 (1982), S. 189-197 
    Details
    ISSN: 0148-7280
    Keywords: egg fusion ; sperm ; capacitation ; fertilization ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In the course of in vitro fertilization of zona-free hamster and pig eggs by boar and guinea-pig spermatozoa it was observed that homologous and heterologous eggs fused together, forming cell hybrids between two or more cells. The fusogenic activity was attributed to spermatozoa and this was the hypothesis tested. The fusogenic activity (coinciding with sperm penetration activity) was dependent on the duration of sperm preincubation, which may be regarded as capacitation in vitro. Fusion occurred only after 3 hr of sperm preincubation and a narrow optimum was detected at 4-4.5 hr. Fusion of eggs was also dependent on sperm concentration. A relatively high proportion of fusions was observed at a sperm concentration of 4.0 × 104 per ml and an optimum was attained at a concentration of 5.0 × 105 per ml. The first fusions were observed at 90 min after semination. After 3 hr more than a half of the eggs reacted, and by 20 hr of incubation 80% of ova were fused. The fusability of eggs was tested and found to occur at 14 hr after ovulation. The fusion process was also studied using transmission electron microscopy. It is supposed that the process of egg fusion may be caused either by a similar mechanism to sperm-egg fusion, or by products released during the sperm acrosome reaction.
    Additional Material: 4 Ill.
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    URL: http://dx.doi.org/10.1002/mrd.1120060302
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    Sperm motility in the horseshoe crab. III. Isolation and characterization of a sperm motility initiating peptide (1982)
    New York, NY : Wiley-Blackwell
    Gamete Research 6 (1982), S. 315-326 
    Details
    ISSN: 0148-7280
    Keywords: sperm ; motility ; peptide ; Limulus ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Sperm motility in Limulus is initiated by a sperm motility initiating factor (SMI) that emanates from Limulus eggs. This report describes the partial purification of SMI (greater than 230-fold purification with respect to protein content) with 40% recovery. SMI appears to be a hydrophobic peptide of 500-2,000 MW. Although probably not purified to homogeneity, SMI is estimated to be active at a concentration of less than 0.2 μM.
    Additional Material: 3 Ill.
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    URL: http://dx.doi.org/10.1002/mrd.1120060404
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    Sperm motility in the horseshoe crab. IV. Extracellular ions and intracellular pH are not mediators of motility initiation (1982)
    New York, NY : Wiley-Blackwell
    Gamete Research 6 (1982), S. 327-342 
    Details
    ISSN: 0148-7280
    Keywords: sperm ; motility ; ions ; pH ; Limulus ; calcium ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Upon dilution into sea water, Limulus spermatozoa undergo a brief flurry of motility (duration 〈 60 sec), after which they are nonmotile until encountering a sperm motility initiating peptide (SMI) that emanates from eggs. Utilizing highly purified SMI extracts and simplified seawater formulations (from which individual ions have been deleted), we found that no specific extracellular ion is required for either dilution-initiated or SMI-initiated motility. Indeed, deletion of one ion (Na+) produced dilution-initiated motility of very long duration (several hours). When motility is initiated by SMI (in normal seawater) there is an increase in intracellular pH (pHi), as indicated by the fluorescent probe, 9-amino acridine; however, this pH, change is not a trigger for motility. As a more general method examining ion movements, the fluorescent probe diS-C3-(5) was used to qualitatively measure changes in the membrane potential of spermatozoa. Although crude SMI extracts caused membrane depolarization, further purification resulted in an almost complete separation of this activity from SMI, thus showing that SMI activation is apparently an electroneutral event. (The membrane-depolarizing factor has a molecular weight 〉 30,000 and does not initiate acrosome reactions.) Experiments utilizing the ionophore A23187 and Ca+2-blocking agents (verapamil and TMB-8) provided tentative evidence that mobilization of intracellular Ca+2 may be required for motility initiation. These results show that neither changes in pHi nor the influx of specific extracellular ions are direct mediators of SMI-initiated motility; however, experiments with pharmacologic agents indicate a possible role for intracellular Ca+2.
    Additional Material: 7 Ill.
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    URL: http://dx.doi.org/10.1002/mrd.1120060405
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    Adenylate cyclase activity in porcine sperm in response to female reproductive tract secretions (1983)
    New York, NY : Wiley-Blackwell
    Gamete Research 7 (1983), S. 169-177 
    Details
    ISSN: 0148-7280
    Keywords: porcine ; sperm ; adenylate cyclase ; phosphodiesterase ; female secretions ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Adenylate cyclase activities were studied in porcine sperm in the presence and absence of Mn++ before and after incubation in vivo and in vitro. Incubation of sperm in vivo for 30 min increased the Mg++-stimulated adenylate cyclase activity from 35.1 pmoles cyclic AMP formed per mg protein per 10 min to 50.4 pmoles. The activity stimulated by Mg++ and Mn++ increased from 392 to 729 pmoles after 30 min of in vivo incubation. Activity after incubation in vivo for 120 min was not different from activity after 30 min. In vitro incubation of porcine sperm in Ca++-free Ringer-fructose resulted in no change, but incubation in oviductal and uterine flushings obtained from gilts soon after ovulation increased Mg++-stimulated activity by 24% and Mg++-+ Mn++-stimulated activity by 49%. In vitro incubations in preovulatory flushings plus follicular fluid or in bovine serum albumin also increased adenylate cyclase activity.
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    URL: http://dx.doi.org/10.1002/mrd.1120070209
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    Association of newly synthesized 3H-arginine-labeled proteins with chromatin structures in fertilization (1984)
    New York, NY : Wiley-Blackwell
    Gamete Research 9 (1984), S. 399-408 
    Details
    ISSN: 0148-7280
    Keywords: sperm ; fertilization ; decondensation ; nuclear protein ; mouse ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Zona-free hamster eggs have been fertilized in vitro with boar spermatozoa in a medium enriched by arginine-3H and the sites of localization of newly synthesized arginine-3H-labeled proteins have been investigated using fine-structure autoradiography. It was confirmed that such proteins are synthesized during fertilization and that they accumulate to a notable degree in decondensing sperm chromatin as well as in the chromatin of the female pronucleus and also of the second polar body. A similar process did evidently take place also in defective pronuclei, characterized by a core of a still condensed chromatin and by remaining nuclear membrane. In such male pronuclei the highest concentration of the label was seen just on the border of the condensed chromatin, on the expected site of nuclear protein exchange. It is supposed that, at least in this experimental system, any morphologically detectable sperm decondensation is accompanied immediately by a shift from sperm basic nuclear proteins to other nuclear proteins.
    Additional Material: 9 Ill.
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    URL: http://dx.doi.org/10.1002/mrd.1120090405
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    Structural components of sea urchin sperm nuclei (1984)
    New York, NY : Wiley-Blackwell
    Gamete Research 9 (1984), S. 469-479 
    Details
    ISSN: 0148-7280
    Keywords: sperm ; nucleus ; histones ; nuclear matrix ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The sea urchin sperm nucleus rapidly loses its conoid morphology and becomes more voluminous and spherical upon its entry into the egg cytoplasm during fertilization. This investigation has attempted to determine what are the structural constraints placed upon the sperm nucleus, so that further investigations might determine the egg cytoplasmic factors that are responsible for modifying nuclear morphology. Isolated sperm nuclei were subjected to various extraction procedures in order to remove the majority of the proteins (histones) and also the DNA; subsequently, the residual structures were processed for and examined by electron microscopy. The data presented in this investigation demonstrate the removal of the sperm nuclear histones plus other nonhistone proteins has no effect on the conoid morphology of the sperm nucleus, yet this protein removal has a profound effect on the structure of the nuclear chromatin. It is also shown that removal of the majority of the nuclear DNA has no effect on the shape of the sperm nucleus. These results indicate that there are other components (possibly a nuclear matrix) associated with the sperm nucleus that are responsible for maintaining its conoid morphology.
    Additional Material: 4 Ill.
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    URL: http://dx.doi.org/10.1002/mrd.1120090410
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    Isolation of the inner acrosomal-nuclear membrane complex from rabbit spermatozoa (1983)
    New York, NY : Wiley-Blackwell
    Gamete Research 7 (1983), S. 215-226 
    Details
    ISSN: 0148-7280
    Keywords: sperm ; cytoplasmic droplet ; membrane ; acrosome ; nucleus ; decondensation ; rabbit ; Bio-Glas 2500 ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Rabbit spermatozoa were passed through a Bio-Glas 2500 (BG) column to yield pure cytoplasmic droplets and consequently the droplet-free spermatozoa. Rapid decondensation of sperm nuclei was achieved by 1 mM dithiothreitol (DTT) and 1.5 M NaCl. This treatment caused removal of the plasma and the outer acrosomal membranes. The viscosity of decondensed nuclei was reduced by pancreatic DNase. Further elution of the suspension from BG column yielded a membrane complex. On the basis of electron microscopic observations and the enzyme analysis, these membranes appeared to be the inner acrosomal-nuclear membrane complex (IANC). The IANC was also prepared from isolated sperm head by DTT-NaCl and Dnasetreatment and low-speed centrifugation.
    Additional Material: 9 Ill.
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    URL: http://dx.doi.org/10.1002/mrd.1120070303
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    Visualization of the F9 antigen on sperm from normal and T/t-complex mutants by immunoscanning electron microscopy (1981)
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 2 (1981), S. 171-183 
    Details
    ISSN: 0192-253X
    Keywords: sperm ; F9 antigen ; T/t-complex ; immunolabeling ; scanning electron microscopy ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The antigens defined by conventional syngeneic antiserum against F9 embryonal carcinoma cells were localized on mature sperm using immunolabeling and scanning electron microscopy. Labeling patterns were compared for normal (+ / +) mice and mice bearing recessive t-haplotypes. The results showed that antigens detected by intact anti-F9 antiserum are expressed similarly in all genotypes, except for sperm from mice bearing the t12-haplotype where the frequency of labeled cells was reduced. Labeling with the IgM fraction of anti-F9 antiserum was lower on sperm from all t-genotypes examined, with sperm from + /t12 males showing the most marked reduction. In all cases, the labeling patterns were similar, and included a labeling of the whole sperm head with complete anti-F9 antiserum and a restriction of the label to the postacrosomal region when the IgM fraction was used.
    Additional Material: 5 Ill.
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    URL: http://dx.doi.org/10.1002/dvg.1020020204
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