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  • 1
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 3 (1983), S. 247-259 
    ISSN: 0886-1544
    Keywords: spermatozoa ; Ciona ; axoneme ; quiescence ; twist ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A simple planar model of sliding can predict the amount of sliding required to form a certain degree of bend. The accuracy of this prediction relies on the assumptions that no twists occur in the axoneme and that no sliding occurs at the base. However, previous studies indicated that twists may occur.This paper explores a new method for quantitating and analyzing twists. Preliminary results using this method showed that there were twists. In order to control for possible artifacts due to fixation and other preparative procedures, the characteristic S-shaped quiescent state of Ciona spermatozoa was studied.Analyses of platinum replicas of those flagella in which this waveform is well preserved suggest that most, if not all, of the twists observed are due to the artifact of a curved shape settling onto a surface. Detailed analyses indicate that if twists do occur in quiescent sperm, they are probably less than 0.4 radian. Since axonemes are evidently easily twisted in rigor, and even after fixation, caution should be exercised in interpretation of axonemal twists.
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Gamete Research 8 (1983), S. 201-218 
    ISSN: 0148-7280
    Keywords: protein kinase ; cyclic AMP ; phosphorylation ; flagella ; motility ; spermatozoa ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Demembranated spermatozoa of Ciona do not become motile when provided with MgATP, unless their motility is activated in vivo before demembranation or unless the demembranated spermatozoa are activated in vitro with cAMP or with the catalytic subunit of a cAMP-dependent protein kinase. CAMP causes a greater than fivefold enhancement of 32P incorporation by demembranated spermatozoa. Analysis by one-dimensional PAGE and autoradiography shows several axonemal protein bands that become 32P-labeled during in vitro activation with cAMP and identifies protein bands whose labeling is specifically reduced if motility of the spermatozoa is activated before demembranation, suggesting that these proteins also become phosphorylated during activation of motolity in vivo. These phosphorylated proteins appear to include dynein heavy-chain components, but axonemal tubulin is not phosphorylated. Partially phosphorylated spermatozoa can be activated by an increase in KCI concentration, which appears to dissociate one phosphorylated component from the axoneme.
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  • 3
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Gamete Research 7 (1983), S. 367-376 
    ISSN: 0148-7280
    Keywords: spermatozoa ; erythrocytes ; epididymis ; proteins ; sperm binding ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Tissue pieces from the caput epididymidis of the rat were incubated in vitro with (35S) methionine to produce radioactive secretory proteins. The radioactive secretory proteins so formed were tested for their ability to bind to washed rat spermatozoa collected from the rete testis and cauda epididymidis, and to rat erythrocytes. The sperm and erythrocytes bound approximately 5% of the total radioactive protein. Binding was protein-specific in that only selected proteins became associated with the cells. Binding was not cell-specific, however, since testicular spermatozoa, caudal spermatozoa, and erythrocytes all bound the same proteins to a similar degree.
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  • 4
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Gamete Research 7 (1983), S. 63-73 
    ISSN: 0148-7280
    Keywords: effect ; spermatozoa ; mouse ; embryo ; development ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The effects on mouse embryo development in vivo of varying the numbers of spermatozoa used in artificial inseminations was studied. The two criteria used in the evaluation of the progress of embryo development were 1) ability to reach the two-cell stage and 2) success of development from the two-cell stage through implantation. A 44% reduction in the yield of two-cell embryos and a 67% reduction in the number of implants was observed when C3HeB/FeJ females were inseminated with one-twentieth the number of spermatozoa estimated to be present in a typical ejaculate. The reduction in the yield of two-cell embryos was substantially reversed by a second insemination of a large number of heat-inactivated spermatozoa 12 hr after the first insemination. The sperm-dependent reduction in development from the two-cell stage through implantation was prevented only by normal viable (unheated) spermatozoa. These results were rationalized by the hypothesis that in female C3HeB/FeJ mice spermatozoa serve physiological functions beyond the fertilization of ova and that spermatozoa may act to foster early embryo development through modulation of the environments embryos experience as they move through the reproductive tract.
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  • 5
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Gamete Research 7 (1983), S. 75-84 
    ISSN: 0148-7280
    Keywords: fate ; spermatozoa ; female ; mouse ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Mechanisms were sought through which the control of preimplantation mouse embryo development by spermatozoa might be effected. A potential route for the transmission of sperm-dependent stimuli to C3HeB/FeJ females was uncovered. It was found that within 24-48 hr after artificial insemination with spermatozoa, in which the DNA had been labeled with tritiated thymidine, a minimum of 9% of the radioactivity was transported across the uterine walls. It was deposited among the maternal tissues in a pattern that differed from the patterns of isotope distribution obtained when either free tritiated thymidine or Escherichia coli cells containing DNA labeled with tritiated thymidine were used instead of labeled sper-matozoa. In sperm-treated animals the ovaries, the adrenals, and a mesenteric lymph node exhibited strikingly large accumulations of radioactivity. The heart, spleen, and uterus manifested lesser accumulations of label, but were higher than liver, kidney, lung, brain, muscle, and intestine. The specific activity of the lymph node was found to decrease during the 12-72-hr period following insemination. This result led to the hypothesis that the lymphatic system could serve as a route for the dissemination, to maternal tissues, of radioactivity originally associated with spermatozoa deposited in the uterus. Heat-inactivated spermatozoa, which have the potential for facilitating the first cleavage of fertilized embryos, exhibited a distribution pattern indistinguishable from untreated spermatozoa. Sperm protein kinase was found to survive the heat inactivation of spermatozoa. This stability was interpreted as being compatible with the kinase functioning as an intermediary in the transmission of sperm-dependent stimuli that control preimplantation embryo development in mice.
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Gamete Research 8 (1983), S. 255-265 
    ISSN: 0148-7280
    Keywords: antigens ; plasma membrane ; intracellular ; monoclonal antibodies ; spermatozoa ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Fluorescent antibody labeling techniques are frequently used to investigate the topography of antigens on spermatozoa. It is generally assumed that these procedures detect molecules only on the sperm surface but we now show that this assumption is not always valid. Using monoclonal antibodies that recognize either surface or internal antigens we demonstrate how spurious conclusions can be made, and we suggest simple procedures for assigning the position of an antigen to the cell surface or to an intracellular organelle. Antibodies against plasma membrane antigens should stain 100% of normal intact spermatozoa, but this proportion should be greatly reduced if the spermatozoa have previously been demembranated. If ≪ 100% of spermatozoa are stained but the proportion increases following permeabilization, then the possibility should be considered that the antigens are intracellular. We conclude that assignment of an antigen to a regional domain on the sperm surface using fluorescent antibody techniques should be validated by a demonstration that the antigen is actually located on the cell surface.
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  • 7
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Gamete Research 7 (1983), S. 401-406 
    ISSN: 0148-7280
    Keywords: epididymis ; secretion ; surface protein ; spermatozoa ; reptilian ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A major soluble protein occurring in two forms, a monomer (protein L) and a polymer, has been identified in the voluminous secretory granules produced by the epithelium of Lacerta vivipara epididymis. An antiserum was raised against protein L and used as an immunohistochemical probe. Indirect immunofluorescence microscopy has indicated that protein L is able to bind to the heads of the spermatozoa. By incubating spermatozoa with buffers of increasing ionic strength and by using nonionic detergents it was not possible to remove completely the protein L. Therefore it was concluded that the binding of protein L to the spermatozoa was not labile and might play an important role in the physiology of the spermatozoa, which is presently under investigation with this convenient model.
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  • 8
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Gamete Research 8 (1983), S. 325-333 
    ISSN: 0148-7280
    Keywords: cytoplasmic droplet ; spermatozoa ; bovine ; crystalloid ; ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: An elongate crystalloid inclusion has been noted in the cytoplasmic droplet of cauda epididymal bovine spermatozoa. The crystalloid appears to be composed of an aggregate of parallel 10 nm diameter filamentous elements that are associated laterally with one another. It has a regular cross-banding pattern that repeats at 13-15-nm intervals. A purified fraction of detached droplets was prepared by centrifugation of sperm suspensions onto Percoll gradients. The detached droplets also exhibited the crystalline inclusion. The origin and possible functions of this structure are discussed.
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  • 9
    ISSN: 0148-7280
    Keywords: spermatozoa ; boar ; crater defect ; electron microscopy ; nucleus ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Nuclear vacuoles resembling the “crater defect” described in bull spermatozoa were observed in 14 boars. Both the incidence of the defect and semen quality were monitored with phase contrast microscopy over a three-month period. The percentages of cratered spermatozoa varied widely both among boars and in ejaculates from the same boar taken on different days. The presence of cratered spermatozoa at a level of 5% or more appeared to be associated with low semen quality. The defect was studied with scanning and transmission electron microscopy and was found to consist of nuclear invaginations, about 0.5 μm in diameter, containing some scanty amorphous electron-dense material. In boars showing a high incidence of spermatozoa with crater defects, abnormalities of the acrosome and perforatorium were common.
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  • 10
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Gamete Research 3 (1980), S. 317-322 
    ISSN: 0148-7280
    Keywords: spermatozoa ; cell surface ; lectin receptor ; autoantigen ; Ricinus ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The isolated rabbit sperm plasma membrane autoantigen RSA-1 has been identified as a receptor for the lectin, Ricinus communis I (RCA). Using purified RSA-1 labeled with125 I, the autoantigen was shown to bind to RCA affinity columns and the eluted fraction bound to specific anti-RSA-1 alloantiserum immunoadsorbent columns.
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  • 11
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Gamete Research 3 (1980), S. 351-367 
    ISSN: 0148-7280
    Keywords: marsupial ; spermatozoa ; nucleus ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Spermatozoa of six species of Australian marsupials have been studied. The nucleus is highly unstable when compared with those of eutherian mammals. When thin films of spermatozoa in buffered saline are air-dried on glass slides, the nucleus disintegrates and flattens, leaving the acrosome, midpiece, and tail intact. This spreading of the nucleus can be inhibited by seminal plasma proteins and by bovine serum albumin, but is potentiated by detergents. The nucleus also decondenses spontaneously in the presence of high concentrations (〉0.25M) of calcium and magnesium salts, leaving the head membranes, acrosome, midpiece, and tail intact. This is inhibited by EDTA. In some species, certain areas of the nucleus appear more resistant t o Ca++/Mg++ treatment, and the initial stages of decondensation are uneven. Ultrastructurally the Ca++/Mg++ dispersed chromatin shows a moderately fine, branching, fibrillar structure, interspersed with dense granules. Treatment with disulphide bond cleaving agents together with detergents results in rapid and complete dispersal of the chromatin and acrosome, and slow digestion of midpiece and tail structures. Treatment with HCl, NaCl, KCl, EDTA, detergents, and sucrose has no effect on nuclear integrity, but treatment with NaOH (0.9-1.0M) results in complete digestion of the whole sperm. These findings are discussed in the light of evolutionary differences between marsupial and eutherian mammals in terms of sperm structure and composition.
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  • 12
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Gamete Research 7 (1983), S. 49-61 
    ISSN: 0148-7280
    Keywords: spermatozoa ; isolation ; Centrifugation ; Percoll ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A procedure using centrifugation in density gradients composed of Percoll was developed for isolation of spermatozoa from mammalian semen. To evaluate the technique, rabbit, human, or bovine semen was layered over continuous Percoll gradients ranging in density from 1.02 to 1.13 gm/ml and centrifuged at 1,500g for 45 min. After centrifugation, the seminal plasma remained above the gradient, whereas the spermatozoa and seminal particles were distributed within the gradient according to their buoyant densities. Unlike most washing techniques, no sperm pellet was formed; instead, the spermatozoa were concentrated into a compact band above the most dense layer of Percoll. The spermatozoa recovered from the gradient were easily resuspended by gentle techniques. Thus, the mechanical stress to the spermatozoa was minimized. Osmotic stress to the spermatozoa was also negligible as the Percoll gradients were isotonic throughout. Spermatozoa obtained by this technique possessed motility equivalent to that of spermatozoa in the unfractionated semen. Sperm suspensions recovered from the gradients contained less than 5% of the nonspermatozoal particles present in the original samples of unfractionated semen. Soluble seminal components were also efficiently removed from the spermatozoa. Thirty billion bovine spermatozoa could be fractionated on a single gradient without loss of effectiveness. Recovery of spermatozoa from these preparative separations averaged 80%. These results demonstrated that Percoll was a superior medium for efficient density gradient isolation of motile spermatozoa free of contamination by other seminal constituents.
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  • 13
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Gamete Research 7 (1983), S. 199-214 
    ISSN: 0148-7280
    Keywords: ultrastructure ; spermatozoa ; nucleus ; Bivalvia ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Sperm ultrastructure and spermiogenesis of the three bivalve species Musculus discors, Nucula sulcata, and Dreissena polymorpha have been studied. During spermatid differentiation in Musculus discors and Nucula sulcata the nucleus attains an elongated rod-like shape. The spermatozoon from Nucula sulcata was found to have a cup-shaped acrosome and five mitochondria surrounding two centrioles in the middle piece. The spermatozoa from Musculus discors has a long complex acrosome. From the distal centriole striated processes extend and attach to the plasma membrane. The spermatozoon of the fresh water species Dreissena polymorpha agrees in all main features with those of other invertebrate groups with external fertilization. It is thus of the primitive type with barrel-shaped nucleus and four to five mitochondria1 spheres in the middle piece. The acrosome is a prominant, complex structure at the apex of the mature spermatozoon.A comparison of sperm ultrastructure among bivalves indicates that there is a certain correlation between the evolution of the elongated sperm nucleus and large, yolk-rich eggs. In species with an elongated sperm nucleus the increased egg size has often led to a lecithotrophic or direct development. The elongated nucleus is a slight modification of the primitive type. There is a great variation in acrosome structure among bivalve spermatozoa, reflecting diverging functional demands at fertilization of the eggs.
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  • 14
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Gamete Research 7 (1983), S. 347-350 
    ISSN: 0148-7280
    Keywords: Taurine-related compounds ; taurine antagonists ; taurine uptake inhibitors ; fertilization in vitro ; hamster ; spermatozoa ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Several taurine-related compounds, taurine antagonists and taurine uptake inhibitors were tested for their effect on hamster sperm motility in vitro. Hypotaurine was approximately three times more effective than taurine. N-methyltaurine and taurocyamine were less effective. Inactive taurine-related compounds were not effective blockers of taurine's spermtimulating activity. However, 1-(4-nitrophenyl)-2-dimethylaminomethyl-l-propenone, a taurine uptake inhibitor, completely suppressed sperm motility at a molar concentration equal to, or less than, that of the taurine added to the incubation medium and also suppressed the motility-sustaining action of the cumulus oophorus.
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  • 15
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Gamete Research 8 (1983), S. 309-323 
    ISSN: 0148-7280
    Keywords: spermatozoa ; Nematoda ; evolution ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The main features of the Nematode sperm cell are the absence of a flagellum and of an acrosome. Transition forms have never been described, as in other animal phyla also reaching the aflagellate condition, like Platyhelminths and Arthropods. The absence of the flagellum must be considered as a definitive acquisition in the group. In addition, centrioles have been demonstrated to be lacking in most cases. The absence of the acrosome is the second general feature of the Nematode sperm cell. Among other features, more or less common to the Nematodes, the most important and general is the presence in the cell periphery of spheroidal membranous vesicles, originated from the Golgi complex but not involved in fertilization or in the production of ascaridin granules. These are absent only in the Ascarid Aspiculuris and the Dorylaimiid Xiphinema, both kinds of sperm having a peculiar shape. These granules are possibly involved in cell motility. Some Nematode sperm have proteinaceous crystalline inclusions originated from the rough endoplasmic reticulum, called ascaridin granules, the role of which remains obscure. A third important feature is the absence of a nuclear envelope, characterizing all described Nematode spermatozoa, the only exception being the Enoplid Mesacanthion, which seems to be for this reason the most primitive model in the group. Other features are the reduced number, or total absence of, the chondriome, an amoeboid movement not owing to an actomyosin system and a dense halo of 10-nm filaments surrounding the perinuclear cytoplasm.In this apparently homogeneous picture, three main evolutionary steps can be recognized. The first one, represented by the primitive Enoploid Mesacanthion, is that of a sperm conserving the nuclear envelope, surrounded by a few mitochondria and many membranous vesicles. The second, the most typical of the group, present in high Enoplida, and in Rhabditida, Strongylida, Ascarida, Spimrida, Trichinellida, is that of roundish, amoeboid spermatozoa devoid of a nuclear envelope but containing mitochondria, membranous vesicles, filaments, microtubules, sometimes centrioles, and sometimes ascaridin granules. The third step is apparently a simplification of the second; in fact, in Tylenchida and Dorylaimiida, the sperm is devoid of membranous vesicles, while in Mononchida and Dioctophymatida it is devoid of mitochondria. Aspiculuris, also devoid of membranous vesicles and having a big mitochondrial derivative, can be assigned to the same level. Nematode sperm evolution does not seem therefore to be a progressive acquisition of new characters, but rather a radiation from an already perfect model of some further simplifications occurring in parallel in most of the orders.
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