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  • Life and Medical Sciences  (1,061)
  • Animals  (908)
  • 1980-1984  (1,969)
  • 1925-1929
  • 1981  (1,066)
  • 1980  (903)
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  • 1980-1984  (1,969)
  • 1925-1929
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  • 1
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 1 (1980) 
    ISSN: 0886-1544
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 1 (1980), S. 1-15 
    ISSN: 0886-1544
    Keywords: centrosomes ; kinetochores ; microtubule initiation ; nuclease enzymes ; electron microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A lysed cell system was used to study the organelle structure and nucleation of exogenous tubulin at kinetochores and centrosomes in mitotic PtK2 cells. We have used this lysed cell system in conjunction with nuclease digestion experiments to determine which specific nucleic acids (DNA or RNA) are involved in either the structure and/or microtubule-initiating capacity of kinetochores and centrosomes. The results indicate that DNase I specifically decondenses the kinetochore plate structure, with the eventual loss in the ability of the chromosomes to nucleate microtubule assembly. DNase I had no effect on either the structure or nucleating capacity of centrosomes. Both RNase T1 and RNase A specifically attacked the amorphous pericentriolar material of the centrosomes, with a concomitant loss in the ability of this material to nucleate microtubule formation. Neither RNase appeared to affect the structure or nucleating capacity of the kinetochore. Therefore, the two types of nucleases appear to exert preferential effects on the different types of microtubule initiation sites in mitotic mammalian cells. The results suggest that DNA is a major component of the kinetochore, while RNA is a major component of the amorphous pericentriolar material. These findings support the concept that microtubule initiation sites in mitotic cells contain nucleic acids which are essential for the structural and functional integrity of the sites.
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  • 3
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 1 (1980), S. 31-40 
    ISSN: 0886-1544
    Keywords: actin ; fascin ; actin cross-linking proteins ; fertilization ; microvilli ; sea urchin eggs ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Following fertilization, the sea urchin egg cortex undergoes a structural change involving the assembly and organization of actin filaments into microvilli. Antifascin localizes this actin cross-linking protein in the microvilli of the fertilized egg cortex but no organized staining is present in the unfertilized cortex. Determination of the actin content of eggs using the DNAase I inhibition assay indicates that actin is about 1.4% of the total protein. Approximately 90% of this actin is soluble in low calcium isotonic extracts of unfertilized eggs while only 60-65% can be recovered in identical extracts of fertilized eggs. Similar measurements for fascin using a radioimmunoassay indicate this molecule represents about 0.3% of the total egg protein, essentially all of which is recovered in low calcium isotonic extracts of unfertilized eggs. After fertilization only 65-70% of this actin cross-linking protein is in the soluble phase. These results demonstrate a markedly different solubility for actin and fascin after fertilization, when the indirect immunofluorescence staining localizes fascin in the microvilli, and are consistent with the idea that fascin organizes newly polymerized actin filaments into the microvillar cores. A consideration of the amounts of actin and fascin incorporated into the cortex after fertilization and the number of microvilli on the egg surface indicates that the measured values are sufficient to account for the observed microvillar elongation.
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  • 4
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 1 (1980), S. 17-29 
    ISSN: 0886-1544
    Keywords: Ca-ion ; Labyrinthula ; contraction ; glycerination ; Ca-reservoir ; cell movement ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Colonies of Labyrinthula, a colonial marine protist, expand by protrusive movements of the specialized slimeways. The movements recorded in time-lapse films are of two types - filopodial and lamellipodial - and occur at rates equivalent to those of cell translocation.Evidence is presented that Ca2+ regulates the contraction of the actomyosin system of filaments present in the slimeways of Labyrinthula. In glycerinated models or in colonies exposed to ionophore A23187 contraction is evidenced by the occurrence of periodic contractions of the slimeways, giving them the appearance of strings of beads. Glycerinated slimeways contract on the addition of Ca2+ and ATP while slimeways provided with ionophore A23187 contract on addition of Ca2+ alone. The concentration required is 1.1 × 10-7 M Ca2+ while concentrations of 6.2 × 10-8 or lower were ineffective. Rates of contraction were measured in time-lapse films which provide evidence that contractions and beading occur everywhere in the slimeway system. When beading occurs, the 6-nm filaments transform from an array of parallel single filaments into an interwoven meshwork.We have identified by pyroantimonate-OsO4 fixation, as possible Ca2+ reservoirs, deposits of Ca2+ in bothrosomes - structures through which cell secretions pass into the slimeways. The electron-dense deposits are located at the base of the bothrosome and disappear after incubation with EGTA. We propose that the translocation of cells as well as the movements of slimeways may be regulated by the cells through the local measured liberation of Ca2+ from the bothrosome where it is sequestered.
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  • 5
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 1 (1980), S. 41-61 
    ISSN: 0886-1544
    Keywords: mitosis ; mitotic spindle ; kinetochore ; microtubule ; micronucleus ; Tetrahymena ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Mitotic micronuclei were isolated from Tetrahymena thermophila and data on spindle ultrastructure were obtained from serial, transverse sections. Comparison of data from nuclei at meta- and early anaphase with data from nuclei at late anaphase showed that during anaphase, sister kinetochores move from the equator to the spindle poles, but kinetochore translocation occurs without any apparent change in either the number or length of kinetochore microtubules. This unprecedented result is ascribed significance with regard to the mechanism of kinetochore transport since there are only a limited number of ways that result could be achieved. The organization of the peripheral sheath changes during anaphase as evidenced by gaps in the sheath at late anaphase. Numerous kinetochore and non-kinetochore microtubules are located in polar regions of the spindle at late anaphase, whereas those regions contained only peripherally arranged microtubules at earlier stages. Tracking of individual kinetochore microtubules in late anaphase nuclei showed that some of them appeared to become incorporated into the peripheral sheath near the pole. At early and late anaphase, crossbridges connect adjacent microtubules throughout the spindle poleward to the kinetochores, as well as in the interzone.
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 1 (1980), S. 63-71 
    ISSN: 0886-1544
    Keywords: Physarum polycephalum ; myosin light chains ; polyacrylamide gel electrophoresis ; calcium ; cytoplasmic streaming ; actomyosin ATPase regulation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Myosin from the slime mold Physarum polycephalum contains three sizes of polypeptides: a heavy chain and two light chains, LC-1 and LC-2. Using a simple qualitative test for calcium binding by comparing electrophoretic migration of the polypeptides in sodium dodecy1 sulfate (SDS) acrylamide gels in the presence and absence of calcium, we have found that Physarum myosin light chain LC-2 migrates with an apparent molecular weight of 16,900 daltons in the presence of the metal ion chelator ethylene glycol bis (B-aminoethyl ether) N,N′-tetraacetic acid (EGTA). However, if calcium chloride is added to the sample prior to electrophoresis, the apparent molecular weight decreases to 16,100. Lanthanide and cadmium ions, but not magnesium, can substitute for calcium. Because the ionic radii of Ca2+, La3+, and Cd2+ are almost identical, we conclude that Physarum myosin LC-2 possesses a very size-specific binding site for calcium. Physarum myosin LC-1 and the heavy chain give no evidence for binding calcium by this test. Since cytoplasmic streaming in the plasmodium of Physarum requires calcium, our evidence indicates that the calcium-binding property of Physarum myosin LC-2 may be important in regulating the production of force by actomyosin in the ectoplasm. Unexpectedly, the myosin light chain in Physarum capable of binding calcium, LC-2, is the essential light chain, while LC-1 is a member of the regulatory class of myosin light chains [V. T. Nachmias, personal communication]. Until now, essential myosin light chains have not been shown to have high affinity divalent cation binding sites. This means a new version of the myosin-based model for actomyosin regulation by calcium may be required to explain cytoplasmic movement in Physarum, and perhaps in other motile systems involving cytoplasmic myosins as well.
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  • 7
    ISSN: 0886-1544
    Keywords: videomicroscopy ; differential interference microscopy ; streaming ; reticulopodial motility ; Allogromia ; microtubules ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A new method called Allen Video-enhanced Contrast, Differential Interference Contrast (AVEC-DIC) microscopy is shown to be sufficiently sensitive to detect several new features of microtubule-related motility in the reticulopodial network of the foraminifer, Allogromia. The method takes advantage of the variable gain and offset features of a binary video camera to operate the DIC microscope under conditions highly favorable for video imaging, but in which the optical image is virtually invisible to the eye yet retains its full information when viewed by a suitable video camera. The improvements are made possible by setting a dé Senarmont compensator to λ/9-λ/4 at maximal working aperture of internally corrected planapochromatic objectives. Under these conditions, the offset feature of the video camera can reject so much stray light from the instrument and specimen that contrast compares favorably with that observed in high-extinction images, and polarizing rectifiers offer scarcely any advantage. Freed from the constraints of the light-limited conditions of DIC microscopy, video images can be recorded 60 times per second, or over 1,000 times the rate of photomicrographs at comparable magnifications under high-extinction conditions.Application of this method to the reticulopodial network of Allogromia has shown that cytoplasmic organelles are translocated only in contact with single microtubules or bundles of microtubules, and that these organelles fail to move when separated from microtubules. Microtubules themselves undergo both axial translatory (“sliding”) and lateral “zipping and unzipping” movements that have been suggested to occur during mitosis and other biological processes.
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  • 8
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 1 (1981), S. 329-347 
    ISSN: 0886-1544
    Keywords: actin ; microfilaments ; heavy meromyosin ; mammary gland ; secretion ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Cytochalasin B, a microfilament-altering drug, inhibits lactose synthesis in lactating guinea pig mammary gland [Biochim. Biophys. Acta 392:20, 1975] but not primarily by inhibiting glucose transport [Eur. J. Cell Biol. 20:150, 1979]. In order to study the possible role of microfilaments in lactose synthesis and secretion, we isolated both the alveolar (milk-secreting) and myoepithelial (contractile) cells from lactating mammary gland. Light microscopy shows that the alveolar cell fraction (viability approximately 71%) is homogenous and that the cells retain strong polarity of secretory structures in the apical region. Two proteins were extracted from the alveolar cell fraction. One (mol wt 42,000) comigrates with skeletal muscle actin on SDS-PAGE gels. The other, a high-molecular-weight (180,000) protein (HMWP) may be analogous to actin-binding protein or clathrin. An extract from the myoepithelial cell fraction also contains a protein that comigrates with actin but no HMWP. Whole tissue extract contains the 42K protein, and a 185K HMWP. Examination of the alveolar cell extract by electron microscopic (EM) negative staining revealed meshworks of multistranded, interconnecting filaments, with attached globular structures (100-200 A) (possibly the HMWP) and single filaments (40-60 A diameter) branching off. To localize these filamentous structures in situ, whole tissue was glycerinated and incubated with rabbit skeletal muscle heavy meromyosin (HMM). Masses of filaments in myoepithelial cells served as convenient standards for HMM decoration. Decorated filaments have cross-arms or projections, unlike the narrow, smooth filaments of control tissue. Decorated filaments in alveolar cells are located beneath the plasma membrane, in close association with secretory vacuoles, and near the Golgi apparatus; filaments near the latter two are often oriented perpendicular to the plasma membrane. Microvesicles are embedded in meshworks under the plasmalemma and near the Golgi apparatus. Intermediate-sized (85-115 A diameter), non-decorated filaments diverge from the meshworks of decorated filaments. Microvesicles are associated with intermediate-sized filaments as well. The association of actin-like filaments with secretory vacuoles and microvesicles and their location in areas of the cell concerned with biosynthetic activities suggest a possible function in the intracellular transport of secretory products.
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  • 9
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 1 (1981), S. 455-468 
    ISSN: 0886-1544
    Keywords: intercellular bridge ; intercellular communication ; cytokinesis ; squid ; ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Incomplete cytokinesis followed by the disappearance of the midbody and spindle remnant results in intercellular bridges between the cells of the blastoderm of the squid embryo. An electron microscope study of the morphology of the stages of development of the intercellular bridge is presented. Cytokinesis ceased as the furrow base reached a diameter slightly larger than the midbody. As furrowing stopped, a dense material accumulated to form a cylindrical sheath 50 nm thick, lining the inner surface of the furrow base. Proteolytic enzymes showed this material to have a significant protein component. As the midbody broke down, vesicles lined the inner surface of the bridge sheath. In this configuration, there was cyto-plasmic continuity between the cells, and organelles appeared to pass through the bridge.The intercellular bridge could become temporarily closed. Vesicles entered the channel and fused with the vesicles lining the inner surface of the sheath. The vesicles enlarged until the channel became occluded with a series of transverse cisternae, the edges of which were embedded in the material of the sheath. When the bridge reopened, the transverse cisterna appeared to dissociate from the sheath, move out of the channel, and break down. Occasionally bridges were seen in which the bridge wall appeared distorted into lobes. It is suggested that such bridges might be in the porcess of breaking down, resulting in the final separation of the cells.
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  • 10
    Electronic Resource
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    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 1 (1981), S. 469-483 
    ISSN: 0886-1544
    Keywords: microtubules ; nucleation ; mitosis ; nocodazole ; immunocytochemistry ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The reassembly of microtubules is described in mitotic cells after release from nocodazole-induced block. The formation of microtubules was followed by light microscopic immunocytochemical staining using the PAP method, combined with to-luidine blue staining of the chromatin. The light microscopic observations on whole cells were compared with ultrastructural observations on thin sections. This step is essential to ascertain complete destruction of microtubules during the nocodazole treatment and to correlate immunocytochemical staining with the presence of microtubules.Removal of nocodazole (10 or 1 μg/ml) after a sufficiently long incubation to induce a complete disappearance of microtubules resulted in the appearance of tubulin staining specifically associated with the centromeres and with one or two isolated points in the cytoplasm. Electron microscopy confirmed that the staining was due to the massive accumulation of small microtubules at the kinetochores and centrosomes. Kinetochore nucleation was seen only in association with condensed metaphase-stage chromosomes and not with the less-condensed prophase chromosomes.In a second type of experiment cells were allowed to enter mitosis in the presence of an incompletely active concentration of nocodazole (0.1 μg/ml). The construction of the mitotic spindle was arrested; however, short microtubules were assembled at the kinetochores and centrosomes.These experiments demonstrate that in living mitotic PTK2 cells the kinetochores, as well as the centrosomes, exert a nucleating action on tubulin assembly.The further elongation of microtubules after removal of nocodazole was seen to occur preferentially along axes between the centrosomes and the kinetochores. This resulted in the construction of normal metaphases that evolved through anaphase and telophase. We have attempted to formulate a hypothesis that may explain the oriented assembly that seems to be essential in the construction of the spindle.
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  • 11
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    Cell Motility and the Cytoskeleton 1 (1981), S. 485-497 
    ISSN: 0886-1544
    Keywords: actin ; tubulin ; nucleotides ; polymerization ; microfilaments ; microtubules ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Both actin and tubulin, the major proteins of the cytoskeleton, bind nucleotide triphosphate (NTP) and exhibit the phenomenon of “polymerization-coupled” NTP hydrolysis. In this report I review the nature of polymerization-coupled NTP hydrolysis, and its possible role in the cellular function of actin and tubulin. Polymerization-coupled hydrolysis may be viewed as simply reflecting differences in the NTPase activity of free subunit as compared to polymer. Making assumptions concerning the values of various rate constants, it is possible to write expressions for the effects of NTP hydrolysis on the kinetics of polymerization. The role of NTP hydrolysis may be viewed in at least three different ways: (1) Hydrolysis alters the kinetics of assembly and disassembly. This leads to a consideration of the role of subunit flow in microtubule and microfilament function. (2) Hydrolysis is an essentially irreversible step that separates the assembly and disassembly reactions. This suggests a role of NTP in the regulation of polymer content during cellular cycles of assembly and disassembly. (3) NTP may allow transient stabilization of intersubunit bonds. This suggests a role of NTP in nucleation and possible regulation of nonequilibrium states of assembly.
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  • 12
    Electronic Resource
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    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 1 (1981), S. 499-515 
    ISSN: 0886-1544
    Keywords: dynein ; tubulin ; axonemes ; microtubules ; microtubule-associated proteins ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Microtubule-associated proteins (MAPs), isolated from brain tubulin, bound to and saturated outer fibers of Chlamydomonas flagella. MAPs present on these microtubules prevented the subsequent recombination of dynein. MAPs also bound to intact axonemes and thus did not specifically bind to the dynein binding sites on the A subfiber. A molar ratio of 1 mole MAP2 per 27 moles tubulin dimers at saturation of the outer fibers with MAP2 suggested that MAPs could effectively interfere with dynein recombination only if the MAPs were near the dynein binding sites to sterically prevent binding. However, electron microscopic observations indicated that MAPs were not localized but, instead, were dispersed around the outer fibers. In addition, MAP2 present at saturating amounts on in vitro assembled brain microtubules had no significant effect on dynein binding. Dynein-decorated microtubules contained clusters of arms suggesting that there may be cooperative interaction between the arms during dynein binding. Because the A subfiber of axonemes contains sites to which dynein preferentially attaches, MAPs may prevent recombination by interfering with cooperative binding to these specific sites. Dynein presumably binds with equal affinity to any protofilament on in vitro assembled microtubules, and, therefore, the MAPs may not be capable of effectively interfering with cooperative binding of dynein to these microtubules.
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  • 13
    Electronic Resource
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    Cell Motility and the Cytoskeleton 1 (1981), S. 167-178 
    ISSN: 0886-1544
    Keywords: nerve growth ; actin ; tubulin ; antibodies ; immunofluorescence ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Embryonic chick nerve cells, from dissociated dorsal root ganglia, were cultured on polylysine substrata and examined for tubulin and actin distribution by indirect immunofluorescence.Antibodies generated against chick brain tubulin produced specific fluorescence in growth cones, neurites, and cell bodies without revealing distribution differences or substructure in the nerve cells. However, at reduced antitubulin concentrations, differences were resolved. Tubulin fluorescence remained uniform and intense in neurites and cell bodies, but exhibited reduced intensity and patterning in growth cones. Nonneuronal cells in the reduced intensity and patterning in growth cones. Nonneuronal cells in the cultures served as controls for typical cytoplasmic tubulin fluorescence distribution. Straining controls demonstrated that fluorescence resulted from tubulin-antitubulin binding.Analogous studies, using antibodies generated against chick brain actin, demonstrated distribution differences at reduced antiactin concentrations, including “hot spots” of intense fluorescence in growth cones and a paucity of fluorescence in neurites.
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  • 14
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    Cell Motility and the Cytoskeleton 1 (1981), S. 237-245 
    ISSN: 0886-1544
    Keywords: centrioles ; symmetry ; triplet blades ; thermal fluctuations ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The paper suggests several principles of construction of a microscopically small device for locating the directions of signal sources in microscopic dimensions. It appears that the simplest and smallest device that is compatible with the scrambling influence of thermal fluctuations as are demonstrated by Brownian motion is a pair of cylinders oriented at right angles to each other. Nine equally spaced blades run in a pitched fashion along the mantle of each cylinder. The blades have a concave cross-section and bend around the circumference of the cylinder in a certain rotational pattern. Considering the striking similarity of this hypothetical device with centrioles, the paper puts forward the conjecture that centrioles locate the direction of hypothetical signals inside cells.
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  • 15
    Electronic Resource
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    Cell Motility and the Cytoskeleton 1 (1981), S. 247-260 
    ISSN: 0886-1544
    Keywords: cilia ; trachea ; ATP-reactivation ; ciliary activity ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Evidence for active sliding of microtubules during ciliary activity has been demonstrated in a number of organisms: sea urchin sperm flagella, protozoan cilia, and mollusc gill cilia. Although there is evidence that active sliding also occurs in mammalian sperm flagella, there is little or no information on whether active sliding of microtubules also occurs in the short (5-μm) cilia of the mammalian trachea or oviduct. Since these cilia are important in tracheobronchial clearance and ovum transport, respectively, it has been important to demonstrate that microtubule sliding is also involved in the activity of somatic cilia. Ciliated apical portions (cortices) and cilia were isolated from rabbit trachea and oviduct, using Triton X-100 to demembranate the cilia. Most of the ciliated cortices reactivated upon addition of ATP, whereas isolated cilia reactivated to a lesser extent. When preparations of cilia were digested with trypsin before or after ATP addition, disintegration of axonemal doublets occurred with about the same frequency as reactivation. These events were recorded using Nomarski optics and dark-field microscopy. When isolated cilia which had been digested by trypsin and exposed to ATP were also prepared for electron microscopy by negative staining, telescoping of doublet microtubules from axonemes could be shown. These results demonstrate that mammalian somatic ciliary doublet microtubules actively slide in a manner similar to that described for invertebrate cilia.
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  • 16
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    Cell Motility and the Cytoskeleton 1 (1981), S. 269-272 
    ISSN: 0886-1544
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
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  • 17
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    Cell Motility and the Cytoskeleton 1 (1981), S. 261-268 
    ISSN: 0886-1544
    Keywords: Tetrahymena ; chemotaxis ; temporal-gradient sensing ; modulation of turning frequency ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The motility pattern of Tetrahymena thermophila in a homogeneous attractant field consists of successive “runs” and “turns.” The turning frequency decreases or increases upon an abrupt increase in attractant or repellent concentration, respectively. The dose-response curve for leucine and methionine yields a saturation curve with half maximum modulation of the turning frequency at a concentration of 15 μM and 2 μM, respectively. The turning frequency is modulated at a threshold concentration of 0.02 μM and 0.50 μM for leucine and methionine, respectively. The decrease (increase) in turning frequency in the presence of an attractant (repellent) jump reverts to prestimulus frequency in a time proportional to the concentration jump. Hence, Tetrahymena seem to employ temporal-gradient sensing for chemotaxis. Spatial-gradient taxis is thus exerted by random walk, which is biased in the direction of the gradient.
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  • 18
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    Cell Motility and the Cytoskeleton 1 (1981), S. 273-273 
    ISSN: 0886-1544
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
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  • 19
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    Cell Motility and the Cytoskeleton 1 (1981) 
    ISSN: 0886-1544
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
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  • 20
    ISSN: 0886-1544
    Keywords: videomicroscopy ; polarization microscopy ; streaming ; reticulopodial motility ; Allogromia ; microtubules ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A new method is described for recording rapid processes of cell motility in polarized light. The Allen video-enhanced contrast (AVEC-POL) method of polarization microscopy achieves significant improvements in resolution, contrast, and the visibility of fine detail by a combination of novel adjustments to a standard (unrectified) polarizing microscope and video camera. Using the full working aperture of a high-power planapochromatic objective lens and compensator setting of λ/9-λ/4, visible images appear lacking in contrast. However, the same images viewed with an appropriate video camera equipped with an electronic offset adjustment can be made to appear with as much contrast as desired, revealing a significantly greater amount of fine detail in the image than can be seen by high extinction visual microscopy alone. At bias retardations between one-ninth and one-quarter wave, the diffraction anomaly observed near extinction disappears. Consequently, polarizing rectifiers are not required with the AVEC-POL method, and images previously requiring photographic exposures of around 20 seconds are sufficiently bright to be registered on the video monitor in 1/60 second. Using an intensity monitor, quantitative measurements of cellular birefringence can be retrieved from live or videotaped images displaying a linear relationship between contrast and phase retardation due to birefringence. The AVEC-POL method also renders accessible to polarized light analysis a number of objects that scatter or depolarize too much light to be studied by high extinction methods. The method is demonstrated on model objects and applied to the highly motile reticulopodial network of Allogromia laticollaris. Rapid motion in close association with microtubules can now be analyzed in greater detail at a significant reduction in the cost of recording.
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  • 21
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    Cell Motility and the Cytoskeleton 1 (1981), S. 303-327 
    ISSN: 0886-1544
    Keywords: cilia ; microtubules ; ATPase ; vanadate ; geometry of sliding ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A dynein arm attachment cycle produces sliding between adjacent doublet microtubules (N and N + 1) of cilia. In intact axonemes, in the absence of ATP, almost all arms appear attached at both ends (rigor). When ATP is added, most arms detach from doublet N + 1. In ATP and vanadate, the arms do not return to rigor, suggesting that ATP hydrolysis is required for re-extension and reattachment of the dynein arm, but not for detachment. Using solutions containing dynein to decorate dynein-less axonemal doublets, we confirm this interpretation. In the absence of ATP, both sides of each doublet decorate with arms. Addition of ATP, ATP and vanadate or AMP-PNP causes immediate arm detachment, but only in the first instance, where extensive ATP hydrolysis can occur, does decoration eventually reappear. Dynein decorates heterologous axonemal doublets and brain microtubules, as well as homologous doublets, suggesting that this mechanochemical cycle may have general applicability in microtubule-based cell motility.
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    Cell Motility and the Cytoskeleton 1 (1981), S. 433-443 
    ISSN: 0886-1544
    Keywords: Physarum ; acellular slime mold ; calcium ion ; calcium-ionophore ; cytoplasmic contraction ; oscillation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Calcium is now generally thought to play a key role in regulating a variety of cellular movements. When the plasmodium of Physarum polycephalum was treated with the calcium-ionophore A23187 or the quasi-ionophore amphotericin B, Ca2+ leaked out. Ca2+ efflux into the ambient solution from the plasmodial strand segment was measured by the luminescence of a photoprotein aequorin, and the tensile force production was recorded simultaneously. Ca2+ efflux oscillated with the same period as the cycle of tension generation in the strand, but the phase of cyclic changes in Ca2+ efflux was opposite to that of tension generation. That is, Ca2+ efflux fell in the increasing tension phase and rose in the decreasing tension phase. Cyclic changes in efflux of Ca2+ are provisionally interpreted as reflecting corresponding changes in concentrations of free Ca2+ in the cytoplasm.
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  • 23
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    Cell Motility and the Cytoskeleton 1 (1981), S. 445-454 
    ISSN: 0886-1544
    Keywords: taxol ; microtubules ; polymerization ; tubulin ; mitotic inhibitor ; protein self-assembly ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Dissociated bovine brain microtubule protein has been shown to reassemble at 0°C in the presence of the drug taxol. Tubulin polymerization was monitored both by electron microscopy of the polymeric structures and by incorporation of tritiated GTP into filterable polymeric structures. Most of the labeled guanine nucleotide uptake into tubulin polymeric structures occurred in the first 30 minutes of incubation with the drug. The initial polymerization event results in the formation of protofilamentous tubulin ribbons. The first microtubules were noted after 1 hour of incubation with the drug. After 20 hours of incubation at 0°C with taxol, the bulk of the polymerized tubulin appeared to be in the form of microtubules. Cold-stable tubulin rings with a mean diameter of 34 nm were present in the reaction mixture before the addition of taxol and throughout the 20-hour incubation. Most of the rings were apparantly not involved in the taxol-induced microtubule assembly. The results are consistant with a model whereby taxol induces an initial formation of protofilamentous ribbon structures, mostly from free tubulin dimers, and a slower subsequent folding of the ribbon structures into microtubules.
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    Cell Motility and the Cytoskeleton 1 (1981), S. 179-192 
    ISSN: 0886-1544
    Keywords: actin ; echinoderm ; fascin ; filopodia ; actin cross-linking protein ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Echinoderm coelomocytes transform from petaloid cells with large motile lamellipodia to filopodial forms. During this morphological transformation, actin filaments extensively reorganize from a random meshwork into tight bundles, which become the skeletons or cores of the filopodia. Antibody localization procedures show that fascin, a 58,000 dalton actin cross-linking protein, becomes incorporated into the filament bundles as they form. Isolated filopodial cores have a pronounced transverse striping pattern, which has been previously identified with fascin crosslinks, and gel electrophoresis identifies a protein in the cores that co-migrates with purified egg fascin. A few of the core fragments also have a distinctive “cap,” which we presume is the membrane insertion site for actin filaments.We have developed a radioimmunoassay for fascin and have used it to study the redistribution of this protein during transformation. Data from the assay indicate that fascin constitutes about 5% of the total cell protein and that substantially more fascin, approximately 1.5-2 times more, is found in the Triton-insoluble cytoskeletons of the filopodial cells than in the petaloid cells. Actin, measured by the DNAase I inhibition assay accounts for approximately 10% of the total cell protein. Approximately 65% of this actin is in a soluble non-filamentous form in the petaloid cells. Our results show that actin polymerization must occur during the cell shape change, since we find approximately 25% more actin in the filopodial cytoskeleton than in the petaloid cytoskeleton. The results show a preferential incorporation of fascin into the cytoskeleton as the cells form filopodia.
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    Cell Motility and the Cytoskeleton 1 (1981), S. 193-203 
    ISSN: 0886-1544
    Keywords: polygonal network ; rat aortic smooth muscle cell ; cell culture ; electron microscopy ; amino acid analysis ; elastin ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Cultured rat aortic smooth muscle cells (SMC) were examined by electron microscopy and found to contain polygonal networks of 75 A° thin myofilament bundles. The cells also had large bundles of longitudinally aligned thin myofilaments with periodically spaced dense bodies. Abundant plasmalemmal vesicles were present at the cell periphery, and the cells were connected by desmosomes. Intercellular spaces contained sparse amounts of elastic fibers, a material generally present in SMC cultures. Analyses of amino acids by automated column-chromatography showed that isodesmosine and desmosine, two amino acid residues unique for elastin, were present. Accordingly, it was concluded that polygonal networks, previously detected solely in cultured nonmuscle cells, were present in SMC.Other findings suggest (1) a change in myofilament arrangement takes place during cell migration, and (2) rat aortic SMC grown in tissue culture flasks is an important experimental tool in the study of cell motility since such myofilament rearrangements were observed to occur up to fourteen days in first passage.
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    Cell Motility and the Cytoskeleton 1 (1981), S. 205-235 
    ISSN: 0886-1544
    Keywords: capping of receptors ; cell locomotion ; cell-surface interactions ; frictional force ; membrane flow ; polymorphonuclear leukocytes ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: As a cell moves over a surface, the distribution of membrane proteins that adhere to the surface will be changed relative to the distribution of these molecules on a static cell. Observations of this redistribution offer, in principle, evidence as to the mechanisms of membrane dynamics during cell locomotion. Toward extracting such information we present and analyze a mathematical model of receptor transport in the membrane by diffusion and convection, as affected by the making and breaking of the bonds between the receptors and the surface as the cell moves.We show that the disruption of receptor-surface bonds at the tail of the cell provides a mechanism by which the frictional force opposing a cell's motion is exerted, and calculate the magnitude of this force as a function of cell velocity. Assuming this to be the major contribution to the frictional force, we show that when the shear force on a cell is above a critical value it is no longer possible for the cell to slide across the surface. For such large forces, it is still possible for the cell to roll; alternatively the cell can be torn free of the surface.Our analysis of existing data on movement of polymorphonuclear leukocytes indicates that cell motion is not accompanied by a bulk flow of membrane from the front to the back of the cell. The data also indicate that cells do not tend to roll as they move over a surface under normal conditions. The data are most consistent with a model where the membrane as a whole is stationary but where receptors that bind to the surface become coupled to sub-membrane contractile proteins.
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    Cell Motility and the Cytoskeleton 1 (1981), S. 387-397 
    ISSN: 0886-1544
    Keywords: birefringence ; polarizing microscope ; sea urchin egg ; cortex ; mitosis ; cleavage ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Birefringence (BR) at the cell surface of fertilized eggs of the sand-dollar, Clypeaster japonicus, during mitosis and cleavage was determined with a photoelectric BR detection apparatus [Hiramoto et al, 1981a]. The cortex of about 2 μm thickness is birefringent positive with respect to the normal to the cell surface. The hyaline layer is negatively birefringent. The halo-layer consisting of a row of microvilli surrounding the egg is positively birefringent in normal Ca-free sea water, while it is negatively birefringent in Ca-free sea water with high refractive index. The BR of the cortex gradually increases over the entire surface during mitosis until the onset of cleavage. The BR of the cortex at the polar region reaches a maximum shortly after the onset of cleavage and then decreases, while the BR of the cortex at the equatorial region begins to decrease shortly before the onset of cleavage, reaches a minimum shortly after the cleavage starts, and then increases again as the cleavage furrow advances. The coefficient of birefringence of the cortex is about 2.5 × 10-5 at the maximum. The BR change of the cortex during mitosis and cleavage is interpreted as a passive deformation caused by the constriction of the contractile ring as well as an active structural change of the cortex occurring in the dividing cell.
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    Cell Motility and the Cytoskeleton 1 (1981) 
    ISSN: 0886-1544
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Cell Motility and the Cytoskeleton 1 (1981), S. 417-431 
    ISSN: 0886-1544
    Keywords: spindle poles ; centrioles ; cell center ; scaffold ; electron microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: I have used fluorescence microscopy and antibodies to 10nm filaments and tubulin labelled with contrasting fluorochromes to compare the distribution of these proteins in endothelial cells during cell division. During interphase the two filament systems have entirely different distributions: The bulk of the 10nm filaments form a ring that surrounds the cell center and nucleus and remains parallel to the substrate, while the microtubules radiate from the cell center to the cell's border. When the mitotic spindle replaces the radial microtubule pattern in mitosis, the spindle poles remain within - and in close proximity to - the ring of 10nm filaments. This was confirmed by electron microscopy which showed the ring and centrioles in the same plane separated by a distance of 300-400 nm.
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    Cell Motility and the Cytoskeleton 1 (1981), S. 399-416 
    ISSN: 0886-1544
    Keywords: myosin heavy chain ; avian muscular dystrophy ; adult and embryonic fast white fibers ; slow red fiber ; rod ; subfragment-1 ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Avian muscular dystrophy is characterized by the degeneration of fast white skeletal muscle fibers, with onset during development. Using a one-dimensional peptide mapping technique, we have detected two forms of the myosin heavy chain in the fast white fibers of adult domestic chickens, one form characteristic of birds homozygous for muscular dystrophy, the other of their normal controls. Four dystrophic strains carrying the same gene for muscular dystrophy were examined.No differences were detected in the embryonic heavy chain peptide maps of normal and dystrophic chickens, consistent with the developmental onset of the condition. Differences were also absent from the peptide maps of heavy chains from slow red fibers, which are unaffected in dystrophy. No dystrophy-specific peptide map differences were detected in the three light chains. Analysis of peptide maps of rod and the heavy chain component of subfragment-1 from normal and dystrophic heavy chains indicates the presence of amino acid sequence differences in the two proteins.
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    Cell Motility and the Cytoskeleton 1 (1980), S. 99-112 
    ISSN: 0886-1544
    Keywords: cell motility ; extracellular matrix ; collagen ; glycosaminogly cans ; collagenase ; hyaluronidase ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The effect of specific components of the extracellular matrix on the motility of tissue cells was studied using organ-cultured aggregates of embryonic fibroblasts. Spherical aggregates of chick embryo heart and skin fibroblasts were fused with [3H]-thymidine-labeled aggregates of the identical cell type. The movement of labeled cells into the unlabeled partner aggregate served as an estimate of cell motility in the cultured tissue-like aggregates. Collagenase treatment decreased the collagen content of heart fibroblast aggregates and increased cell motility; ascorbic acid treatment increased the collagen content of skin fibroblast aggregates and decreased cell motility. Reduction of the glycosaminoglycan content with testicular hyaluronidase had no measurable effect on cell motility in heart fibroblast aggregates.
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    Cell Motility and the Cytoskeleton 1 (1980), S. 73-97 
    ISSN: 0886-1544
    Keywords: nematodes ; muscle structure ; mutants ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A search for new mutants with altered body-wall muscle cell structure has been undertaken in the nematode C elegans. One-hundred seventeen mutants were isolated after mutagenesis with ethyl methanesulfonate or ultraviolet light, enrichment by a motility-requiring test, and screening by polarized light microscopy; 102 of these mutants were in ten previously established genes, whereas 15 mutants permitted the identification of seven new complementation groups in C elegans. Two of the new genes map on linkage group I (unc-94 and unc-95) and four genes are sex linked (unc-96, unc-97, unc-98, and unc-99). One complementation group (unc-100) could not be mapped because of the special characteristics of its cohort mutants. Representative mutants of the mapped genes were examined by polarized light and electron microscopy. All of the mutants exhibit disruptions of the normal A and I band organization of thick and thin filaments. Several of the mutants produce collections of thin filament-like structures. In one of these cases, HE177 demonstrated collections of somewhat wider, intermediate-sized filaments as well, and the HE195 mutant produces paracrystalline aggregates of thin filaments amidst looser arrangements of similar structures. The mutants in newly identified genes, as well as the new mutants in previously established genetic loci, have promise as tools in the study of myofibrillar assembly and function. Among the 22 complementation groups associated with body-wall structure in C elegans, it is likely that some genes code for regulatory and morphogenetic functions in addition to the well-studied structural, contractile, and calcium-associated proteins in muscle.
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    Cell Motility and the Cytoskeleton 1 (1980), S. 113-129 
    ISSN: 0886-1544
    Keywords: tubulin ; Drosophila ; β-ecdysterne ; differentiating ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Drosophila Kc cells exposed to physiological doses of the moulting hormone, β-ecdysone, elongate, become motile, and subsequently aggregate. This pattern of morphogenesis was found to require the assembly of a microtubular cytoskeleton. Tubulin content was significantly increased in hormone-treated cells when compared to controls, as measured by a 3H-colchicine-binding assay. However, determinations of rates of tubulin synthesis and breakdown revealed no difference between control and hormone-treated cells for either parameter. When tubulin content was assayed by methods that do not depend on colchicine-binding activity, no difference between hormone-treated and control cells was observed. These results are discussed in terms of a model in which β-ecdysone affects the distribution of tubulin in “assembly-active” and “assembly-inactive” pools.
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    Cell Motility and the Cytoskeleton 1 (1980), S. 159-162 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Cell Motility and the Cytoskeleton 1 (1980), S. 131-140 
    ISSN: 0886-1544
    Keywords: sea urchin coelomocytes ; motility ; filopodial formation and elongation ; ciné film analysis ; scanning electron microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Sea urchin coelomocytes were examined during their morphological transformation from petaloid to filopodial forms by scanning electron microscopy and ciné film analysis. Petaloid coelomocytes have a variable morphology but, in general, consist of numerous thin sheets of cytoplasm, the petals, arranged in three dimensions around a central nuclear region. The transition to the filopodial form can occur in either substrate-attached or suspended cells and begins with the formation of several microspikes at the edge of each petal. These become more apparent as the cytoplasm between each microspike/filopodium is retracted centripetally. Concomitantly, the diameter of the flattened cell is increased by as much as twofold as the filopodia actively lengthen at a uniform, average rate of 0.5 μm/minute. The transformation process requires ca 15 minutes and is complete when the cell diameter no longer increases. These filopodia are functionally distinct from the passively produced retraction fibers observed in cultured mammalian cells. The formation of filopodia is biphasic and includes both a cytoplasmic retraction phase and an active extension phase.
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    Cell Motility and the Cytoskeleton 1 (1980), S. 141-157 
    ISSN: 0886-1544
    Keywords: axon guidance ; chemotaxis ; haptotaxis substrate pathways ; development ; pattern biology ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: In multicellular organisms, guidance cues are either diffusible molecules or cellular or extracellular surfaces that are found in reproducible locations and that orient migrating cells and cell processes. The pattern of the guidance cues usually determines the complex in vivo migration routes of motile cells and cell processes. Within organisms, guidance cues are found to be organized in two general patterns: (a) broad gradients - such as diffuse chemotactic gradients; (b) discrete routes (substrate pathways) - such as chemotactic gradients confined to long channels, and such as the axon surface which represents a long specific highway for migrating Schwann cells.
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    Cell Motility and the Cytoskeleton 1 (1980), S. 163-163 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Cell Motility and the Cytoskeleton 1 (1980), S. 167-167 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Cell Motility and the Cytoskeleton 1 (1981) 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Cell Motility and the Cytoskeleton 1 (1981), S. 349-362 
    ISSN: 0886-1544
    Keywords: Ca2+ ; flagella ; symmetry ; vanadate ; spermatozoa ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Increased Ca2+ concentration causes a reversible increase in asymmetry of the flagellar bending waves of “potentially symmetric” demembranated sea urchin spermatozoa. When these flagella are immobilized with 5 μM vanadate, increased Ca2+ concentration causes a reversible increase in the total bend angle between the tip and the base of the immobilized flagella. These effects of Ca2+, and the movement which can be activated by CaATP2-, can be inhibited by vanadate, but in both cases, high concentrations of vanadate, of the order of 100 μM, are required. These observations suggest that ATP, possibly in the form of CaATP2-, is required for the Ca2+-induced change in shape of the flagella, but other observations suggest that the magnitudes of asymmetry and total bend angle are more closely related to Ca2+ concentration than to CaATP2- concentration.
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    Cell Motility and the Cytoskeleton 1 (1981), S. 363-370 
    ISSN: 0886-1544
    Keywords: Ca2+ ; Mg2+ ; symmetry ; flagella ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Potentially asymmetric spermatozoa are obtained when spermatozoa are demembranated in the presence of a low Ca2+/Mg2+ ion concentration ratio. They swim with asymmetric bending waves even when reactivated at low Ca2+ concentrations, and become more asymmetric when Ca2+ is increased. Potentially symmetric spermatozoa, which swim with symmetric bending waves at low Ca2+ and become asymmetric as the Ca2+ is increased, can be obtained by exposing the flagella to a high Ca2+/Mg2+ ratio, either during or subsequent to demembranation. The rate of this conversion is an increasing function of temperature and Triton concentration. Potentially symmetric spermatozoa can be reconverted to potential asymmetry, if the exposure to high Ca2+/Mg2+ is brief, and is terminated by addition of EGTA and Mg2+ before diluting the spermatozoa. The conversion to potential symmetry may involve removal of a labile component from the axoneme.
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    Cell Motility and the Cytoskeleton 1 (1981), S. 371-385 
    ISSN: 0886-1544
    Keywords: rotating filaments ; cytoplasmic streaming ; Nitella ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Our knowledge about the actin-containing characean filaments on the basis of light and electron microscopical investigations is reviewed. Dynamic filamentous networks, known already from isolated droplets, were detected in Nitella rhizoidal cells using light microscopical techniques. Earlier light microscopic observations in cytoplasmic droplets are confirmed and complemented by new model experiments with rotating helices. The motile phenomena occurring at the filament bundles (ring formation, wave propagation, particle translocation, net dynamics, rolling motions, formation of side arms) can, in this way, be imitated in detail. Thus, the concept of cytoplasmic streaming as a translocation along bundles of rapidly rotating helical filaments is supported. In order to explain unidirectional cytoplasmic streaming, a periodic winding up and unwinding of fine filaments is postulated by which ions are periodically bound and displaced. The formation of side arms which is favored during unwinding results in a screw-mechanical different behavior of the filaments in the two directions of rotation and therefore causes permanent particle transport in one direction.
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    Gamete Research 4 (1981), S. 333-362 
    ISSN: 0148-7280
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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    Gamete Research 4 (1981), S. 463-472 
    ISSN: 0148-7280
    Keywords: oocyte ; meiotic competence ; estrogen ; PMSG ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Oocytes were removed from the follicles of rats at 15 to 31 days of age, and their ability to resume meiosis (“meiotic competence”) in vitro was correlated with their diameter and the stage of follicular development. The majority of oocytes explanted on day 15 did not resume meiosis when placed in culture, but the percentage of competent oocytes increased from 14.1% ± 3.0% on day 20 to 67.6% ± 3.3% on day 26 of age. This ability to resume maturation correlated well (r = 0.98) with the increase in diameter of oocytes and coincided with the development of antral follicles.Hypophysectomy on day 15 of age, but not on day 20, reduced the percentage (P 〈 0.001) and number (P 〈 0.001) of competent oocytes and was accompanied by a reduction in diameter of oocytes. Treatment with PMSG or E2 increased the number (P 〈 0.001) and percentage (P 〈 0.001) of competent oocytes. These results suggest that the ability of oocytes to mature in vitro is dependent upon stimulation by gonadotropins and that this action of gonadotropin may be mediated by production of estrogen within the follicles.
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    ISSN: 0148-7280
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Gamete Research 4 (1981), S. 486-486 
    ISSN: 0148-7280
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    ISSN: 0148-7280
    Keywords: oocyte-cumulus complex ; granulosa cell ; monolayer formation ; LH/FSH effects on progesterone ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Oocyte-cumulus complexes and granulosa cells were harvested from small (1-2 mm), medium (3-5 mm), and large (6-12 mm) porcine antral follicles and cultured for 2 and 3 days. The effects of various doses of purified hCG and human FSH on progesterone secretion and monolayer formation were examined. After a 2-day culture period it was found that FSH was more effective in stimulation of progesterone secretion by cultured oocyte-cumulus complexes than in granulosa cells harvested from small follicles (P 〈 0.01), whereas hCG was more effective in stimulating progesterone secretion in granulosa cells than in oocytecumulus complexes harvested from large follicles. In contrast, after a 3-day culture period, granulosa cells secreted more progesterone compared to oocytecumulus complexes under control conditions or in the presence of hCG or FSH. After 3 days both FSH and hCG stimulated progesterone secretion by oocytecumulus complexes and granulosa cells; however, the hormone effect was greater upon granulosa cells than oocyte-cumulus complexes. After 3 days of culture in the case of both follicular cell types, there was a greater response to FSH in the case of cells harvested from small compared to large follicles. The reverse was true in the case of hCG responsiveness.Monolayer formation ability of oocyte-cumulus complexes was greater in the case of complexes harvested from small and medium than complexes harvested from large follicles. Addition of hCG to the cultures led to a dose-dependent decrease in monolayer formation by oocyte-cumulus complexes harvested from all sizes of follicles.
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  • 49
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    Gamete Research 4 (1981) 
    ISSN: 0148-7280
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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  • 50
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    Gamete Research 4 (1981), S. 499-506 
    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.
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  • 51
    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.
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  • 52
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    Gamete Research 4 (1981), S. 525-533 
    ISSN: 0148-7280
    Keywords: capacitation ; fertilization ; spermatozoa ; in vitro ; Little Brown Bat ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The temperature dependence of capacitation in bat sperm (Myotis lucifugus lucifugus) was studied by monitoring fertilizations rates of zona-free hamster ova at different temperatures. Spermatozoa were cultured in BWW medium at temperatures 4°C, 24°C, 32°C, 42°C, and 55°C from 0-24 hr. Activation of sperm could be determined visually due to the change in movement seen through light microscopy. Activation was later confirmed by higher rates of fertilization. Preincubation of the bat sperm was found to have a direct effect on the success of penetration of the zona-free hamster ova. Holding bat spermatozoa at low temperature for long intervals allowed them to remain motile but unable to fertilize. Sperm are not irreversibly damaged, however, and activation, when the temperature is increased to 32°C, is faster than when sperm are intitially put at 32°C, resulting in good fertilization rates.
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  • 53
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    Gamete Research 4 (1981), S. 535-545 
    ISSN: 0148-7280
    Keywords: mouse ; embryo ; zygote ; electron microscopy ; reproductive physiology ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Initial changes in the fine morphology of apparently normal mouse zygotes and embryos were studied in serial sections of mouse oviducts that had been fixed in situ. These changes included extra sperm in the perivitelline space, nuclear budding, the presence of large nucleoli, perinuclear vesicles, the concentration of cytoplasmic organelles, and the extrusion of cytoplasmic ground substance. Initial changes in the nucleus and cytoplasm were undetectable when the zygotes and embryos were examined with the light microscope. It was concluded that serious abnormalities in zygotes and embryos, which may not be identified at the light microscope level, may be detected if they are examined in the electron microscope. Therefore, zygotes and embryos should be critically evaluated before being rated as normal.
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  • 54
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    Gamete Research 3 (1980), S. 121-132 
    ISSN: 0148-7280
    Keywords: sperm nucleus ; fish oocyte ; germinal vesicle (GV) ; nuclear formation ; chromosome ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The processes occurring from sperm penetration to chromosome formation in the cytoplasm of Oocytes matured in vitro, after removal of the germinal vesicle (GV) and before hormonal stimulation, were observed with electron microscope. The dechorionated oocytes, matured without the participation of the GV material, responded to sperm penetration by initiating a cortical reaction within 20 seconds after insemination. The pentrating sperm nuclei transformed to male pronuclei with vesiculation of the nuclear membrane, chromatin decondensation, and formation of a pronuclear membrane. Before cleavage, however, no chromosome formation was observed in these oocytes. Instead, the fully grown pronuclei change to a picnotic chromatin mass without or with an only fragmented nuclear membrane, then disappeared. On the contrary, sperm nuclei that penetrated into the cytoplasm of naked eggs containing GV material during maturation underwent pronuclear and chromosomal formation. Judging from these observation in Oryzias oocytes, the GV material seems to be unnecessary for the formation of pronucleus from the compact sperm nucleus, but is essential for the process of chromosomal formation.
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  • 55
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    Gamete Research 3 (1980), S. 179-202 
    ISSN: 0148-7280
    Keywords: Sertoli cell ; spermatogenesis ; junction ; germ cell ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 56
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    Gamete Research 3 (1980) 
    ISSN: 0148-7280
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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  • 57
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    Gamete Research 3 (1980), S. 203-209 
    ISSN: 0148-7280
    Keywords: actin ; mitochondrial movement ; spermiogenesis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The presence of actin filaments around mitochondria during vertebrate spermiogenesis was demonstrated by immunofluorescence and immuno-electron microscopy and by heavy meromyosin decoration. The presence of actin is supposed to be related to mitochondrial rearrangements occurring in the spermatid stage.
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  • 58
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    Gamete Research 3 (1980), S. 133-140 
    ISSN: 0148-7280
    Keywords: rat oocytes ; maturation ; oxygen consumption ; cumulus cells ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Oocytes collected from immature PMSG-treated rats on the morning of proestrus were allowed to mature in culture either surrounded by their cumulus cells or after denudation. It was found that the time course of oocyte nuclear maturation was similar whether the cumulus cells were present or not. The oxygen consumption of noncultured oocytes was 0.12 nl/hr/oocyte and increased by 40% after four to eight hours in culture with intact cumulus. Respiration of oocytes cultured without cumulus remained constant throughout the culture, except for a transient decrease after four hours.It is concluted that the cumulus cells do not affect the spontaneous nuclear maturation in vitro, but that the metabolism in oocytes cultured with intact cumulus is different from that of cultured denuded oocytes. Furthermore, it appears that the rise in oocyte oxygen consumption is not a prerequisite for nucler maturation.
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  • 59
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    Gamete Research 3 (1980), S. 141-148 
    ISSN: 0148-7280
    Keywords: cow blastocysts ; zona pellucida ; stability and location of antigenic material ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Labeling of the zona pellucida of cow blastocysts with zona-specific anti-serum shows that antigenicity is unaffected by abnormal cleavage, in vitro culture, or frozen storage. The uniform labeling in thin sections indicates that the zona pellucida is homogeneous antigenically. Heavier labeling of the inner and outer surfaces of the zona pellucida in thick sections appers to be due to greater porosity of these regions, in which the zona material becomes highly dispersed, or even partly solubilized, thereby permitting the formation of an antigen-antibody matrix.
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  • 60
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    Gamete Research 3 (1980), S. 169-177 
    ISSN: 0148-7280
    Keywords: oviduct ; oviductal fluid ; mucin ; steroids ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Utilizing the intra-abdominal flask technique to collect oviductal fluid, the presence of two or possibly three reproductive-tract-specific antigens have been observed in rabbit oviductal fluid. Two of these antigens may be accounted for by the two forms of uteroglobin. The other antigen has a molecular weight greater than 200,000 daltons and its concentration in oviductal fluid is under hormonal control. During pseudopregnancy (PSP), when progesterone concentrations are high, or upon progesterone administration, the concentration of this high molecular weight antigen doubles in oviductal fluid. This correlates well with the previously observed increase in release of secretory products from the oviductal epithelia.
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  • 61
    ISSN: 0148-7280
    Keywords: Bryophyta ; Phaeoceros ; spermatid morphogenesis ; spermatogenesis ; ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: An ultrastructural examination of spermatogenesis in Phaeoceros has shown nucleoli to be present in spermatogenous cells and to persist until the centrioles become associated with nuclei of young spermatids. At the onset of multilayered structure (MLS) formation, well-defined aggregations of osmiophilic strands begin to form in the nuclei of young spermatids and disappear shortly after chromatin condensation starts in the midstage spermatids. When the centrioles in the young spermatids are orientated perpendicular to the nuclear envelope, the nucleoplasm immediately in front of them is densely stained. Where the spline tubules of the MLS extend over the nucleus, the nuclear envelope is devoid of pores, and the inner nuclear membrane is contacted internally by the local deposition of dense staining nucleoplasm. Chromatin condensation begins with strands extending perpendicularly from the dense staining nucleoplasm beneath the spline and continues with the nuclear beak becoming filled with condensed chromatin. As the MLS lamellae disappear acropetally, the rear portion of the anterior mitochondrion (AM) extends back under the nuclear beak which now narrows to a size that approximates the anterior end of the nucleus of a spermatozoid. By the end of the mid-spermatid stage, the nucleus has coiled approximately one gyre of a helix and the five or six central slpine tubules extend over the plastid which is now located beneath the front end of the AM. Several profiles of endoplasmic reticulum confluent with the nuclear envelope are present. Possible factors which might play a role in determining the morphology of the mid-spermatids are discussed.
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  • 62
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    Gamete Research 3 (1980), S. 211-216 
    ISSN: 0148-7280
    Keywords: acrosome ; human sperm ; lectin ; capacitation ; fertilization ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Because the acrosome of human sperm is too small to be directly visualized by phase-contrast microscopy, acrosome reactions (that is loss of the acrosome) are generally not evaluated in studies of human sperm capacitation and fertilization. Nevertheless, it would be useful in such studies to have a technique for easily identifying and quantitating acrosome-reacted sperm. In this paper, we describe a method for labeling the human sperm acrosome with fluorescein-conjugated Ricinus communis agglutinin-60 (FITC-RCA); we show that in sperm without acrosomal caps, FITC-RCA labeling occurs either not at all or only in the equatorial segment of the acrosome. To determine if the absence of FITC-RCA labeling in the acrosomal cap region gives a reliable estimate of acrosome reactions, washed sperm or sperm incubated in a capacitating medium (BWW) were divided into two groups, which were then fixed for FITC-RCA labeling or transmission electron microscopy. Counts of acrosome reactions made by each method were similar, and we observed an increase in the percentage of reactions following incubation in BWW. We conclude that the FITC-TCA labeling technique is a reliable method for accurately scoring the percentage of acrosome-reacted human sperm.
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  • 63
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    Gamete Research 4 (1981) 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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  • 64
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    Gamete Research 4 (1981), S. 1-1 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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  • 65
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    Gamete Research 4 (1981), S. 3-13 
    ISSN: 0148-7280
    Keywords: oocyte maturation ; parthenogenesis ; preimplantation development ; gonadotropins ; mouse ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Of eggs ovulated in LT/Sv mice, 10-20% undergo spontaneous parthenogenetic activation, and 40-50% of the parthenotes develop to blastocysts when cultured in simple defined medium from the one-cell stage. Similar percentages of oocytes isolated from Graafian follicles undergo parthenogenetic activation after spontaneous maturation in simple defined medium, but embryonic development proceeds no further than the two-cell stage. The simple defined medium that supported preimplantation development of ovulated eggs and spontaneous maturation of extrafollicular oocytes contained no serum, free amino acids, or vitamins. The present experiments were conducted to determine what conditions during spontaneous maturation of extrafollicular oocytes could promote the ability of oocytes to develop to blastocysts after parthenogenetic activation and mimic the environment of preovulatory follicles.Cumulus-enclosed oocytes that were matured in simple medium supplemented with fetal bovine serum (FBS) developed to blastocysts after spontaneous parthenogenetic activation. Furthermore, minimum essential medium (MEM), a complex medium containing free amino acids and vitamins, could substitute completely for FBS for maturing oocytes from (C57BL/6J × LT/Sv)F1 mice, and to a lesser extent for maturing LT/Sv oocytes. Therefore, even though germinal vesicle breakdown in mouse oocytes and preimplantation development of mouse eggs can occur in the absence of an exogenous supply of free amino acids and vitamins, a complete, or normal, mouse oocyte maturation cannot. These results also demonstrated that gonadotropins are not necessary during oocyte meiotic maturation for parthenogenetically activated eggs to develop through the preimplantation stages.Luteinizing hormone or 17β-estradiol in MEM during oocyte maturation had no effect on the subsequent development of parthenotes. In contrast, follicle stimulating hormone (FSH) and progesterone in the maturation medium decreased the number of ova that subsequently cleaved, and FSH decreased the number of cleaved eggs that developed to blastocysts.
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  • 66
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    Gamete Research 4 (1981), S. 35-40 
    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.
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  • 67
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    Gamete Research 4 (1981), S. 41-47 
    ISSN: 0148-7280
    Keywords: zona pellucida ; antibody ; sperm receptors ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Mouse zonae pellucidae contain receptors for sperm throughout their structure since spermatozoa will bind to both the inner and outer surfaces of isolated zona fragments. Antibodies raised against mechanically isolated mouse zonae pellucidae were only capable of suppressing sperm binding to the outer zona surface in association with the formation of a precipitate in this region.These results indicate that such antisera are not capable of interacting directly with the sperm receptors on the zona pellucida but rely upon the less efficient mechanism of steric hindrance to prevent sperm from gaining access to the sperm binding sites.
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  • 68
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    Gamete Research 4 (1981), S. 57-63 
    ISSN: 0148-7280
    Keywords: taurine ; hypotaurine ; spermatozoa ; in vitro fertilization ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Taurine and hypotaurine were examined for their efficacy in replacing sperm motility factor (SMF), prepared from bovine adrenal cortex, for in vitro fertilization in the golden hamster. Combinations of these amino acids at concentrations of 0.001, 0.01, 0.1, and 1 mM together with 16 μM isoproterenol (a catecholamine β-agonist) were added to the sperm incubations. After three hours of sperm preincubation, oviductal eggs were added to the sperm suspensions and examined for penetration and stage of fertilization after three or five hours of culture. At 0.001 mM, neither taurine or hypotaurine was capable of maintaining motility of hamster sperm for four to 4½ hours or of inducing fertilization. With all other concentrations, both amino acids were found to maintain motility of sperm as well as SMF. Hypotaurine stimulated motility to a greater extent than taurine and both required isoproterenol for the greatest motility. A low proportion of cumulus-free ova were fertilized when sperm were preincubated with either amino acid alone over the range of 0.01 to 1 mM; however, over 80% fertilization was consistently obtained when isoproterenol was also present during sperm incubation. Proportions of ova fertilized with taurine or hypotaurine present during sperm preincubation were comparable to those achieved with SMF. The possibility that taurine or hypotaurine is the sperm motility factor is discussed.After three hours of sperm/egg incubation, a lag in the early events of fertilization was observed in experimental groups treated with one of the amino acids (0.01 mM) alone compared with groups treated with isoproterenol present. However, if sperm/egg incubation was extended from three to five hours, no increase in number of eggs penetrated was found. Therefore, the delay observed at three hours was considered a function of fewer numbers of capacitated sperm present in the absence of isoproterenol rather than of the need for an extended capacitation time.
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  • 69
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    Gamete Research 4 (1981), S. 121-131 
    ISSN: 0148-7280
    Keywords: preimplantation mouse embryos ; DNA synthesis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: This paper describes a sensitive, reproducible, and automated procedure to measure DNA synthesis in preimplantation mouse embryos. Conditions for the DNA synthesis assay have been optimized as follows: (1) 4 μCi/ml3H-thymidine (sp. act. 20 Ci/immole); (2) a labeling period from 2 to 7 hours; (3) a 3-hour preincubation period for blastocysts and from 0 to 7 hours preincubation for 8-cell embryos; and (4) from 1 to 64 embryos per assay. The amount of DNA synthesis per embryo was found to be directly proportional to the number of cells (nuclei) per embryo. The described assay should be useful for future studies on the effect of synthetic and natural compounds on the development of preimplantation mouse embryos, as measured by perturbations in embryonic DNA synthetic activity.
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  • 70
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    Gamete Research 4 (1981), S. 139-149 
    ISSN: 0148-7280
    Keywords: selenium ; rat sperm ; mitochondrial capsules ; cysteine ; protein ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The keratinous capsules surrounding rat sperm mitochondria were isolated 24 days after intratesticular injections of [75Se] selenite or [35S] cysteine. Dodecyl sulfate-polyacrylamide gel electrophoresis of purified, doubly labeled mitochondrial capsules revealed only a single 75Se-labeled component, whose molecular weight was 17,000, in agreement with previously reported observations obtained with cruder sperm fractions. Most of the 35S label and the major zone of stained protein on the gels coincided with the position of 75Se, suggesting that selenium is associated with a cysteine-rich structural protein. The level of selenium in rat sperm, 195 ± 3.2 ng/108 sperm (approximately 30 ppm), determined by hydride generation and atomic absorption spectrophotometry, is consistent with a structural function for this trace element in the sperm.
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  • 71
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    Gamete Research 4 (1981), S. 185-192 
    ISSN: 0148-7280
    Keywords: spermiogenesis ; sperm abnormalities ; sterility ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Purkinje cell degeneration (pcd) is a neurological mutation in the mouse that causes male sterility, but not female sterility. In order to assess the effects of this mutation on spermiogenesis, the structure of the testis and of epididymal spermatozoa was examined by transmission and scanning electron microscopy. In the mutant males, the sperm count was reduced, sperm were nonmotile, and 93% of the sperm were characterized by structural abnormalities of the head, the tail, or both. In the testes of mutant mice, Sertoli cell structure was normal, as were also the early stages of spermiogenesis. However, the elongating and maturing spermatids were characterized by abnormally shaped heads and tails with extraneous and ectopic outer dense fibers. These defects were common in the testes of the mutant mice and rare in the testes of the littermate control mice. It was concluded that the structural abnormalities of the pcd sperm occurred during spermiogenesis and were not due to degeneration of the sperm in the epididymis. These structural abnormalities are similar to those found in all other reported male sterile mutants of the mouse; therefore, although they are caused by the expression of the pcd gene, they are not unique to the expression of this gene.
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  • 72
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    Gamete Research 4 (1981), S. 219-229 
    ISSN: 0148-7280
    Keywords: fertilization ; sea urchin ; Arbacia punctulata ; Erythrosin B ; dyes ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Fertilization and ionophore activation of the sea urchin Arbacia punctulata were inhibited in the presence of six analogs of the dye fluorescein. The concentration of any one dye needed for blockage of sperm or Ca-ionophoremediated activation in 50% of the eggs (I50) was a function of the dye's lipid solubility. Substantially higher concentrations of each dye were required to block activation by Ca-ionophore (A23187) than were needed to inhibit sperm activation. A detailed study of the action of Erythrosin B (tetraiodofluorescein) showed that its effects were readily reversible. The I50 concentration of Erythrosin B increased as temperature increased from 10 to 25°C. The kinetics of blockage indicated that Erythrosin B blocked some early step in the program of fertilization. The results suggest that these anionic dyes may inhibit fertilization by preventing successful sperm-egg fusion.
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  • 73
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    Gamete Research 4 (1981), S. 241-250 
    ISSN: 0148-7280
    Keywords: spermatozoan movement ; insect; axoneme ; accessory tubule ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In the mosquito Aedes notoscriptus nine accessory tubules develop from the doublets of the axoneme whose single central fibre forms late in spermiogenesis. The 9 + 9 + 1 mosquito sperm-tail propagates two waves of differing frequency and amplitude. The structural basis for the movement is of interest.Mosquito sperm were demembranated until all movement ceased and were reactivated with a solution containing adenosine triphosphate (ATP). They immediately moved in a normal fashion. Like untreated sperm, they always propagated double waves. This indicates that the plasma membrane does not control the double wave.After sonicating sperm treated as above, the high-frequency, low-amplitude waves sometimes propagated alone beyond a damaged region of sperm-tail, whereas the large-amplitude, low-frequency waves never propagated alone beyond a damaged region. This suggests that the latter waves are generated by an anterior site, possibly the centriole or centriole adjunct. The structure that propagates the large amplitude wave is not known; possibly the accessory tubules may slide against the axonemal doublets.Any region of the sperm-tail is capable of generating and propagating the low-amplitude, high-frequency wave, for which the structure responsible is probably the axoneme.
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  • 74
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    Gamete Research 4 (1981) 
    ISSN: 0148-7280
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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  • 75
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    Gamete Research 3 (1980) 
    ISSN: 0148-7280
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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  • 76
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    Gamete Research 3 (1980), S. 1-7 
    ISSN: 0148-7280
    Keywords: mouse ; in vitro fertilization ; inhibitors of fertilization ; calcium ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Better than 75% fertilization of C57BL/6 mouse eggs with C57BL/6 sperm is obtained in vitro in a modified Kreb's-Ringer-bicarbonate medium containing 8 mM HEPES. No fertilization of obtained when Ca2+ is omitted from this medium. The drug verapamil, which interferes with Ca2+ channels and blocks the acrosome reaction [Schackmann et al, 1978] and fertilization in the sea urchin, also blocks fertilization of mouse eggs in vitro when included in complete medium at a concentration of 80 μg/ml. Tetraethylammonium, which inhibits delayed axonal potassium currents and prevents the acrosome reaction in sea urchin sperm, also completely inhibits fertilization of mouse eggs in vitro at a concentration of 5 mM. Tetramethylammonium, which does not inhibit potassium movements at the same concentration reduces fertilization by about 50%. The data are consistent with the hypothesis that ion movements are necessary for activation of the sperm and/or egg in mouse fertilization.
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  • 77
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    Gamete Research 3 (1980), S. 17-24 
    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.
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  • 78
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    Gamete Research 3 (1980), S. 45-57 
    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.
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  • 79
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    Gamete Research 3 (1980), S. 247-257 
    ISSN: 0148-7280
    Keywords: spermatozoa (salmonid) ; adenosine triphosphate (ATP) in sperm motility ; cyclic adenosine monophosphate (cAMP) in sperm motility ; reactivation of trout sperm motility ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The motility of salmonid spermatozoa initiated by dilution of the milt with ovarian fluid or isotonic saline is brief duration; it was believed that it can be activated only once in the life of the spermatozoon. Dilution of the milt with an equal volume of isotonic saline (0.12 M-NaCl) containing 5 mM-3-isobutyl-1-methylxanthine (MIX) prolonged and intensified sperm motiliy. When motility had stopped after initial mobilization with saline or ovarian fluid, it could be reactivated by addition of MIX; reactivated spermatozoa fertilized eggs. Dilution with saline containing K+ (24 mEq/liter) did not initiate sperm motility even in the presence of MIX. The spermatozoa were mobilized by subsequent with 0.12 M-NaCl. The concentration of adenosine triphosphate (ATP) in sperm suspensions dropped on dilution with saline and rose as motility ceased, but declined without subsequent recovery following dilution with MIX-saline. The concentration of cyclic adenosine monophosphate (cAMP) rose and fell sharply on initiation of motility and rose again after motility had declined. While salmonid spermatozoa can be mobilized by dilition with saline alone, the effectiveness of MIX in reactivating “spent” spermatozoa supports the assumption that cAMP plays a role in the initiation of sperm motility.
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  • 80
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    Gamete Research 3 (1980), S. 279-290 
    ISSN: 0148-7280
    Keywords: ovoperoxidase ; zona hardening ; zona pellucida ; mouse eggs ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: One consequence of fertilization or parthenogenetic activation of mammalian eggs is an altaration in the solubility proprieties of the zona pellucida, known as zona hardening. Several lines of evidence indicate that an ovoperoxidase, which is activated and/or secreted from mouse eggs. Following parthenogenetic activation, corss-links tyrosine residues in the zona pellucida and results in hardening of the zona. First, zona hardening, as determined by decreased solubility of the zona in pronase, is inhibited by several compounds known to inhibit peroxidases. Inhibitors of hardening include phenylhydrazine, sodium sulfite, sodium azide, and glycine ethyl ester. Second, tyrosine analogs inhibit zona hardening, unless the phenolic hydroxyl group or ortho position is blocked. That is, O-methyltyrosine (methyl substitution of phenolic hydroxyl) does not inhibit hardening; o-methyltyrosine (methyl substitution of one ortho position) partially inhibits, whereas tyramine and N-acetyltyrosine (free hydroxyl and ortho positions) effectively block hardening. Finally, exogenous horseradish peroxidasepromotes limited hardening of the zona in unactivated eggs. These results are consistent with a peroxidase catalyzed cross-linking of tyrosines in the zona that results in hardening of the zona pellucida.
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  • 81
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    Gamete Research 3 (1980), S. 291-298 
    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.
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  • 82
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    Gamete Research 3 (1980), S. 307-307 
    ISSN: 0148-7280
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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  • 83
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    Gamete Research 3 (1980), S. 305-306 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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  • 84
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    Gamete Research 3 (1980), S. 309-316 
    ISSN: 0148-7280
    Keywords: fertilization ; membrane potential ; bonellin ; amino acid incorporation into proteins ; DNA synthesis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We here describe further studies on the action of bonellin on sea-urchin eggs. Bonellin brings about Some of the changes that are known to occur in the egg upon fertilization. In particular, it appears to cause the increased rate of incorporation of amino acids into proteins, the increase of the voltage noise, and the exocytosis of some of the cortical granules. A comparison with the effect of ammonia is discussed.
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  • 85
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    Keywords: female meiosis ; estrogen synthesis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: It has been suggested that the production of estrogens by the fetal ovary may modulate the entry or progression of meiosis in the female mammalian fetus. In the present study the possibility that the site of this steroid synthesis is the rete ovarii system was explored in the fetal mouse of gestational ages 12.5 to 18 days. The method of ultracytochemical localization of 3β hydroxy-steroid ferricyanide was used. Reaction product was found in the cytoplasm of the rete ovarii (prefollicular) cells as early as day 14 with increasing amounts seen at later gestational ages. The presence of this essential enzyme system in cells closely applied to oogonia and oocytes during an active meiotic period must be considered in developing concepts of meiotic entry.
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  • 86
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    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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  • 87
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    Gamete Research 3 (1980), S. 369-377 
    ISSN: 0148-7280
    Keywords: yolk ; preimplantation embryo ; ultrastructure ; hamster ; mouse ; rat ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Yolk material of preimplanation stages of embryos of the hamster, mouse, and rat were examined by a standardized electron microscopical procedure. The material was encountered as fibrils, scattered more or less densely in the cytoplasm. In the hamster, the material was present in large masses and the fibrils had a chain-like appearance when cut longitudinally. The ultrastructure of the fibrils was compatible with a helical pattern. The fibrils had a width of about 40 nm and the pitch (the axial distance of the repeating unit) was about 30 nm. In the mouse, the yolk material was dispersed in the cytoplasm forming small plaque-like groups. Also, in this species the fibrils were chain-like but smaller than in the hamster. The fibrils were often closely situated, resulting in images with varying crystalline appearances. In the rat, the yolk appeared as light areas occupying a substantial part of the cytoplasm. The fibrils in the yolk plaques were sparse and diffusely outlined. They were thinner than the fibrils of the mouse-yolk material, did not display any helical pattern at the resolution used, but showed a periodicity.
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  • 88
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    Gamete Research 3 (1980), S. 405-406 
    ISSN: 0148-7280
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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  • 89
    ISSN: 0148-7280
    Keywords: sperm-zona contact ; fertilization ; peptides ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: An investigation was made as to the nature of two of the factors, termed S1, released within the first 30 minutes after contact is made between capacitated hamster sperm and the zona pellucida in vitro. Previous studies showed that these S1 factors were detected two and 20 to 25 minutes after the gametes were combined and that, based on filtration studies, the former possessed a molecular weight of less than 5,000 daltons. The present results show that the quantity of the 20-25-minute S1 factor released into the supernatant increased linearly as a function of the sperm concentration. This factor passed unimpeded through a filter with a 5,000 molecular weight cutoff but only 42% of the activity traversed a filter with a cutoff of 2,000 daltons. The two-minute S1 factor, in the virtual total absence of cells, was stable for 10 to 15 minutes, but lost significant activity upon longer incubation. Under the same conditions, the 20-25-minute factor lost approximately 25% of its activity within 15 minutes, but remained stable at this level for at least 45 minutes of incubation. Both S1 factors were not affected by a mixture of glycosidases, but were inactivated by subtilisin, trypsin, and leucine aminopeptidase which was contaminated with endopeptidases. The activity of the two-minute S1 factor appeared more susceptible to the action of the proteases than that of the 20-25-minute S1 factor. In contrast to previous results obtained with the two-minute S1 factor, the release of the 20-25-minute S1 factor was not inhibited by the inclusion of soybean trypsin inhibitor a t concentrations which are known to inhibit penetration of the zona by the sperm. The results suggest that the two- and 20-25-minute S1 factors are peptides which are not identical.
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  • 90
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    Gamete Research 4 (1981), S. 15-23 
    ISSN: 0148-7280
    Keywords: secretory IgA ; oviduct fluid ; acrosin ; spermatozoa ; fertilization ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A major inhibitor of acrosin in rhesus monkey and rabbit oviduct fluid, isolated by isoelectrofocusing in sucrose gradients, displayed a broad peak in the acidic region of the column and was demonstrated to contain secretory IgA specific for acrosin. Its identity was established by immunodiffusion, by the removal of acrosin inhibition with antisera to IgA (α-chain), and by its correct molecular weight during ultracentrifugation. Purified human serum IgA also inhibited rabbit, rhesus monkey, and human acrosins, but neither purified human IgG nor IgM had any inhibitory effect on these acrosins. Neither oviduct fluid secretory IgA nor purified human serum IgA inhibited the activity of bovine pancreatic trypsin. The high specificity of secretory IgA for acrosin and its presence in every rabbit and rhesus monkey oviduct fluid specimen examined suggests a possible regulatory role for this antibody in reproduction.
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  • 91
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    Gamete Research 4 (1981), S. 25-34 
    ISSN: 0148-7280
    Keywords: fertilization ; pronuclei ; chromatin decondensation ; protamine ; kinase ; glutathione reductase ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In order to derive information about possible mechanisms by which the sperm head is converted into the male pronucleus during fertilization in the rabbit, unfertilized egg homogenate was assayed for two enzyme activities. Protamine was extracted from rabbit sperm, purified, and labelled with [14C] in an in vitro reaction and used as a probe to assay for a protein kinase which could transfer [32P]PO4 from [γ-32P]ATP onto the substrate. A kinase with a pH optimum of approximately 8.0 to 8.5 is described. Assays for the enzyme glutathione reductase were performed using homogenates from eggs or embryos at three early stages of development. Results suggest that oocytes can oxidize 2.58 × 10-6 μmol NADPH per minute per oocyte, unfertilized eggs 5.16 × 10-7 μmol NADPH per minute per ovum, and 20- to 24-hour postcoitus fertilized eggs 2.30 × 10-6 μmol NADPH per minute per ovum. The relevance of these observations to male pronuclear formation is discussed.
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  • 92
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    Gamete Research 4 (1981), S. 49-56 
    ISSN: 0148-7280
    Keywords: meiotic maturation ; progesterone ; mianserine ; oestradiol ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Maturation of Xenopus laevis oocytes can be induced by mianserine, a tricyclic antidepressant. K+-free medium facilitates this maturation process. Mianserine must be kept in contact with the oocytes during the whole process of maturation for maximal efficiency. It is inactive after injection into the oocytes. Mianserine induces the formation of maturation-promoting factor (MPF) in the treated oocytes. Mianserine-induced maturation is strongly inhibited by theophylline, even in K+-free medium. Progesterone displays synergistic effects with mianserine for the induction of maturation. Likewise, oestradiol shows cooperative maturing effects with progesterone as well as with mianserine. It is suggested that mianserine exerts its primary effects on oocyte maturation by inhibiting a membrane adenylate cyclase.
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  • 93
    ISSN: 0148-7280
    Keywords: spermatozoon-crayfish (Astacus astacus) ; ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The star-like spermatozoon of Astacus astacus consists of a spheroidal central body around which various prolongations of same, denominated spines, are arranged. In the interior of the gamete the following parts may be distinguished: (1) The acrosomic region, formed by a complex vesicle, or thick-walled, helmetshaped body, whose opening is orientated towards the nuclear region. In the interior of the vesicle different structures can be appreciated. (2) The nuclear region, which is formed by a large cupuliform nucleus limited by a double membrane. In the nucleoplasm numerous bundles of microtubules, mixed with noncondensated chromatin fibers, are found. (3) The laminar region, present in other Decapoda, is practically nonexistent. Within the spines of these spermatozoa, only microtubules can be observed. The morphology of this crayfish is similar to that presented by Brachiura, another group of Reptantia.
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  • 94
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    Gamete Research 4 (1981), S. 83-95 
    ISSN: 0148-7280
    Keywords: human spermatozoa ; flagellum ; motion ; microcinematography ; kinematics ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A microcinematographic (50 f/s) study was performed on motile human spermatozoa. Eighty percent were found to have a linear trajectory and a pseudo-sinusoidal head displacement pattern. Throughout their progression, the spermatozoa periodically rotated on their longitudinal axis at a frequency equal to that of flagellar wave formation. These waves were found always to begin on the same side of the cell and to propagate in the flattened plane of the head until the moment of rotation. At this time the wave had reached a point near the middle of the flagellum. Beyond this point, the flagellum moves out of the plane of the head. Different variables used to characterize the movement of spermatozoa included the velocity of progression, amplitude and velocity of head displacement, frequency of rotation, wave amplitude, and propagation velocity of the flagellar wave. Among these variables, it was the propagation velocity of the wave that was found to be best correlated with the velocity of spermatozoan progression. This flagellar wave exhibited two stages, one of initiation and one of propagation.
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  • 95
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    Gamete Research 4 (1981), S. 113-119 
    ISSN: 0148-7280
    Keywords: human spermatozoa ; morphology ; semen analysis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Human spermatozoa were treated with the detergents Hyamine 2389, Triton X-100, or sodium dodecyl sulphate (SDS) and surface replica preparations and Papanicolaou-stained smears made. From the ultrastructural studies it was found that Hyamine caused a dissolution of the plasmalemma to reveal the outer acrosomal membrane that was often absent over the anterior cap region; whereas Triton, in addition, caused severe damage to the entire acrosome. Treatment with SDS rapidly cleaved the tails from the majority of the spermatozoa, stripped the plasmalemma from both the head and tail regions of the cells, and also removed the whole of the outer acrosomal membrane plus most of the acrosomal contents. However, when viewed in the Papanicolaou-stained preparations, the spermatozoa were all of apparently normal morphology.These studies make it clear that some caution must be used when interpreting human sperm morphology assessments made under the light microscope. While it is not suggested that the more involved technique of surface replication should be used in routine semen analyses, it does represent a valuable technique for use with research material.
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  • 96
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    Gamete Research 4 (1981), S. 133-138 
    ISSN: 0148-7280
    Keywords: inhibition of fertilization ; bovine zonae pellucidae ; passive immunization ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Passive immunization with antiserum prepared against isolated bovine zonae pellucidae inhibited fertilization in the cow. The minimum dosage of antiserum (titer 27 by immunofluorescence) required for complete inhibition was 2 ml/kg of body weight.
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  • 97
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    Gamete Research 4 (1981), S. 151-169 
    ISSN: 0148-7280
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 98
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    Gamete Research 4 (1981) 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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  • 99
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    Gamete Research 4 (1981), S. 171-183 
    ISSN: 0148-7280
    Keywords: mouse ; spermatogenesis ; sperm differentiation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A high frequency (42%) of sperm from the inbred homozygous mouse strain PL/J are abnormal. Head shape abnormalities occur in 15% of the total sperm; and 27% of the sperm are headless, with the mitochondria condensed into a mass at the caudal end of the midpiece region. The sperm without heads exhibit relatively normal motility. Electron microscopy of the testes indicates that some of the abnormal sperm in PL/J males result from a failure of the paired centrioles to attain a normal position on the nucleus opposite the acrosome prior to implantation, or to attach at all. The centrioles that are not attached to the nuclear envelope can differentiate to form the principal piece and midpiece region. The frequency of headless variants in heterozygous F1 indicates that the trait is mainly recessive. The offspring from the backcross of the F1 to homozygous PL/J parents did not give a clear-cut segregation pattern. The frequency of abnormal sperm in the F1 and the backcross is higher when the female parent is a PL/J.
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  • 100
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    Gamete Research 4 (1981), S. 193-202 
    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.
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