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  • 1
    Electronic Resource
    Electronic Resource
    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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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Journal of Morphology 179 (1984), S. 95-114 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Quantitative lateral and dorsoventral cineradiography shows that the masticatory movements of the mandible, condyles, tongue, and hyoid of Pteropus giganteus (Chiroptera) move along highly regular paths that are characteristic for each of the three food types tested.Mandibular movements are predominantly orthal, although a small forward translation occurs early in opening and small lateral deflections occur in both opening and closing phases. These deflections are related to the existence of active (bolus bearing) and balancing sides of the jaws, chewing being not truly bilateral. The deflections are associated with a shift of both condyles toward one side. In consequence the active condyle is located in a lateral part of the associated fossa, the inactive condyle in a medial part. Food transfer from side to side involves a reversal of the chewing direction during opening. Such reversals are especially frequent near the end of a chewing sequence.The fore, middle, and hind parts of the tongue differ in their movement patterns. Movements of the fore part, and to a lesser extent of the middle part, follow the open-close movements of the lower jaw. The hind part of the tongue moves predominantly dorsally during slow closing and ventrally during fast opening and fast closing. All three parts move forward during slow closing and slow opening, and backward during fast opening and fast closing. Movements of the hyoid are closely synchronized with those of the hind part of the tongue. Furthermore, tongue and hyoid movements are synchronized with jaw movements. All cycles of Pteropus giganteus are transport cycles, and the synchrony appears to reflect the consistency of the food (soft pulp, juices). Food consistency also accounts for the high swallowing rate and the absence of any significant difference between nonswallowing and swallowing cycles.
    Additional Material: 11 Ill.
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  • 3
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 2 (1982), S. 497-508 
    ISSN: 0886-1544
    Keywords: cilia ; electric motor control ; ciliates ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: We have studied the motor responses to membrane hyperpolarization of the marginal cirri in Stylonychia using voltage-clamp, high-speed cinematography, and computer-processing techniques. The cirri started beating when voltage step amplitudes rose beyond 5 mV. The power stroke was oriented toward the posterior cell and (hyperpolarizing motor activation). The frequency rose slightly during a voltage step, and decreased with similar rates for 100 ms following the step end. Amplitude and duration of the step tended to increase the motor response of the cirri. The late response declined exponentially. The time constant of the decay rose with the step amplitude. Among three response parameters tested (frequency, duration, number of cycles), the number of evoked ciliary cycles was best correlated with the amplitude of the hyperpolarization. Comparisons with the responses to depolarizing voltage steps reveal similarities in the relaxation of ciliary activity which appears to be uncoupled, in part, from the electric membrane events during the voltage stimulus.
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  • 4
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 2 (1982), S. 205-210 
    ISSN: 0886-1544
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Additional Material: 2 Ill.
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  • 5
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 2 (1982), S. 483-496 
    ISSN: 0886-1544
    Keywords: Cilia ; Ca ; motor control ; ciliates ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: We have studied quantitative aspects of ciliary motor responses to membrane depolarization in the ciliate Stylonychia using voltage clamp and high-speed cinematograhpy techniques and employing computer-processing methods for evaluation. Depolarizations beyond 4 mV activate the cirri (compound cilia) which are at rest in the absence of a stimulus. The power stroke of activiated cirri is oriented toward the cell anterior. The frequency and duration of beating increase with rising depolarization. With very large positive stimuli (≥ 150 mV) activation of the response is delayed until the end of the voltage step (“off-response”). The peak frequecy is essentially unaltered during sustained depolarization. The frequency drops exponentially following repolarization of the membrane. The time constant of the decay in ciliary activity rises with the amplitude, not with the duration of the depolarization. The ciliary motor response is most adequately represented by the number of evoked ciliary cycles (ciliary work), and appears to be related to the amplitude of the depolarization.
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 4 (1984), S. 443-468 
    ISSN: 0886-1544
    Keywords: actin ; microfilaments ; HMM ; phagocytosis ; cytochalasin ; Paramecium ; fluorescence microscopy ; electron microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Using heavy meromyosin (HMM) or the fragment S1 of myosin as probes for actin microfilaments, we studied their organization in Paramecium both by fluorescence and electron microscopy.In interphasic cells, HMM decorates (a) most prominently the periphery of nascent and young food vacuoles and their route during the early phase of their intracellular transit; (b) a thin meshwork radiating from the gullet throughout the cytoplasm; (c) a small area beneath the pore of contractile vacuoles and beneath the cytoproct when open to release food residues. Most of these HMM-decorated structures are in close contact with microtubular arrays. All HMM decoration disappears in dividing cells and in cytochalasin-treated cells. In vivo, the drug immediately blocks food vacuole formation but does not affect cytokinesis, cyclosis, contractile vacuole pulsation, defecation, or nuclear movements.The data show that, as in the cells of other organisms, actin microfilaments form defined arrays that undergo physiologically controlled cycles of assembly/disassembly. These arrays contribute (at least in the phagocytotic process) to diverse types of movement: constriction, membrane fusion, and migration of food vacuoles. However, aside from their massive concentration along the phagocytotic tractus, actin microfilaments are neither major structural components of Paramecium cytoplasm nor the only cytoskeletal components ensuring motility or contractility processes.
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  • 7
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Gamete Research 10 (1984), S. 423-432 
    ISSN: 0148-7280
    Keywords: lectins ; germ cells ; gametogenesis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Fluorescent lectins were used to study the chemical nature of carbohydrate moieties present on the surface of female and male germ cells isolated from mouse gonads during fetal and early posnatal development. Concanavalin A (ConA), lens culinaris agglutinin (LCA), ricinus communis agglutinin (RCAI) and wheat germ agglutinin (WGA) bound intensely to the germ cell plasma membrane at all stages studied. Other lectins such as ulex europaeus agglutinin (UEAI) and agglutinin (SBA) did not bind or bound moderately (SBA to female germ cells only).Distinct developmental-related changes were observed when female germ cells were labeled with fluorescein-conjugated peanut agglutinin (PNA) or dolichos biflorus agglutinin (DBA). DBA and PNA binding was absent or weak in fetal female and male germ cells, but became intensely positive in oocytes in the immediate postnatal period. The percentage of oocytes stained with DBA increased during the first three days after birth, and from day 3-4 onwards all oocytes were strongly labeled.I suggest that these changes in lectin binding reflect changes in biochemical structure of the oocyte surface related to differentiative events occurring in the mouse ovary immediately after birth.
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  • 8
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Gamete Research 8 (1983), S. 309-323 
    ISSN: 0148-7280
    Keywords: spermatozoa ; Nematoda ; evolution ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The main features of the Nematode sperm cell are the absence of a flagellum and of an acrosome. Transition forms have never been described, as in other animal phyla also reaching the aflagellate condition, like Platyhelminths and Arthropods. The absence of the flagellum must be considered as a definitive acquisition in the group. In addition, centrioles have been demonstrated to be lacking in most cases. The absence of the acrosome is the second general feature of the Nematode sperm cell. Among other features, more or less common to the Nematodes, the most important and general is the presence in the cell periphery of spheroidal membranous vesicles, originated from the Golgi complex but not involved in fertilization or in the production of ascaridin granules. These are absent only in the Ascarid Aspiculuris and the Dorylaimiid Xiphinema, both kinds of sperm having a peculiar shape. These granules are possibly involved in cell motility. Some Nematode sperm have proteinaceous crystalline inclusions originated from the rough endoplasmic reticulum, called ascaridin granules, the role of which remains obscure. A third important feature is the absence of a nuclear envelope, characterizing all described Nematode spermatozoa, the only exception being the Enoplid Mesacanthion, which seems to be for this reason the most primitive model in the group. Other features are the reduced number, or total absence of, the chondriome, an amoeboid movement not owing to an actomyosin system and a dense halo of 10-nm filaments surrounding the perinuclear cytoplasm.In this apparently homogeneous picture, three main evolutionary steps can be recognized. The first one, represented by the primitive Enoploid Mesacanthion, is that of a sperm conserving the nuclear envelope, surrounded by a few mitochondria and many membranous vesicles. The second, the most typical of the group, present in high Enoplida, and in Rhabditida, Strongylida, Ascarida, Spimrida, Trichinellida, is that of roundish, amoeboid spermatozoa devoid of a nuclear envelope but containing mitochondria, membranous vesicles, filaments, microtubules, sometimes centrioles, and sometimes ascaridin granules. The third step is apparently a simplification of the second; in fact, in Tylenchida and Dorylaimiida, the sperm is devoid of membranous vesicles, while in Mononchida and Dioctophymatida it is devoid of mitochondria. Aspiculuris, also devoid of membranous vesicles and having a big mitochondrial derivative, can be assigned to the same level. Nematode sperm evolution does not seem therefore to be a progressive acquisition of new characters, but rather a radiation from an already perfect model of some further simplifications occurring in parallel in most of the orders.
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  • 9
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Gamete Research 5 (1982), S. 125-135 
    ISSN: 0148-7280
    Keywords: nicotine ; sea urchin eggs ; electrical events ; polyspermy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have studied some of the effects of nicotine on sea urchin eggs, spermatozoa, and their interaction using electrical recording techniques and fertilization-rate experiments. Pretreating eggs with nicotine enhances the fertilization rate, whereas this drug has an inhibitory effect on spermatozoa. Pulse-treated eggs or eggs fertilized in the presence of nicotine give rise to attenuated step depolarizations, which may be attributed to a decrease in membrane resistance (Rm) of the egg or, in the latter case, to an alteration to the spermatozoon. Concurrently, with the change in the step depolarization there is a reduction in amplitude of the fertilization potential (FP) suggesting that the cortical reaction is in some way altered. Nicotine has no effect on the Rm of fertilized eggs or oocytes, where there are no cortical granules. We suggest that nicotine alters the cortex of sea urchin eggs-possibly by causing a partial dissolution of cortical granules-which renders the eggs more receptive to spermatozoa. The reductions in amplitude of the step depolarization and the FP are consequences of this alteration.
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  • 10
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    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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