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  • 1985-1989  (1,389)
  • 1985  (1,389)
  • Life and Medical Sciences  (1,389)
  • 1
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 5 (1985), S. 1-15 
    ISSN: 0886-1544
    Keywords: contractile non-actin filaments ; dinoflagellates ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The motility and fine structure of the marine planktonic dinoflagellate Kofoidinium and members of other related genera have been investigated. Several types of shape change were found to occur: slow morphogenetic changes (which also occurred as restitution movements in response to injury), movements associated with the ingestion of food and the evacuation of wastes, and more rapid movements concerned with the capture of prey. All these movements seemed to be brought about by the contraction of refractile tracts within the cytoplasm, which were found to contain 2.3-nm filaments, some with a complex striated appearance. This and other evidence suggests that these filaments, which have counterparts in many other protists, are not actin filaments.
    Additional Material: 22 Ill.
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 5 (1985) 
    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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  • 3
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 5 (1985), S. 175-193 
    ISSN: 0886-1544
    Keywords: primary cilia ; connective tissues ; secretory organelles ; extracellular matrix ; cybernetic probe ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: More than 300 primary cilia have been identified electronmicroscopically in a variety of embryonic and mature connective tissue cells. To further define the enigmatic function of these cilia, we examined the interrelationships between the basal apparatus and cytoplasmic organelles and the ciliary shaft and the extracellular matrix. The basal diplosome was consistently associated with the secretory organelles including the maturing face of the Golgi complex, Golgi vacuoles and vesicles, the microtubular network, the plasma membrane, and coated pits and vesicles. Small vesicles and amorphous granules were also observed within the ciliary lumen and adjacent to the ciliary membrane. Microtubule-membrane bridges linked axonemal tubules to the ciliary membrane. The position, projection, and orientation of the axoneme were influenced by the structural organisation and mechanical properties of the matrix and frequently caused angulation of the ciliary shaft relative to the basal body. Located midway between the secretory apparatus and the extracellular matrix, primary cilia would appear ideally situated to mediate the necessry interaction between the cell and its surrounding environment prerequisite to the formation and maintenance of a functionally effective matrix. We propose that primary cilia in connective tissue cells could act as multifunctional, cellular cybernetic probes, receiving, transducing, and conducting a variety of extrinsic stimuli to the intracellular organelles responsible for effecting the appropriate homeostatic feedback response to changes in the extracellular microenvironment.
    Additional Material: 17 Ill.
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  • 4
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 5 (1985), S. 239-249 
    ISSN: 0886-1544
    Keywords: tektins ; microtubules ; flagella ; cilia ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Affinity-purified antibodies against Strongylocentrotus purpuratus sperm flagellar tektin polypeptides have been tested for cross-reactivity with microtubules isolated from various sources, using indirect immunofluorescent staining and antibody binding to nitrocellulose replicas of SDS polyacrylamide gels. The antitektins reacted with sperm tail axonemes from four genera of sea urchins and with cilia from sea urchin embryos. Antibody binding was observed only if the specimens were prefixed by methods that would not preserve them well at an ultrastructural level. However, even after such fixation regimes, no antibody binding was detected to cytoplasmic microtubule arrays in the same embryos, to mitotic spindles isolated from sea urchin or to gill cilia from a mollusc. We conclude that, if tektins are present in sea urchin egg cytoplasmic microtubules, they are sufficiently different from the sperm tektins to have no common strongly antigenic determinants.
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  • 5
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 5 (1985), S. 333-350 
    ISSN: 0886-1544
    Keywords: eel sperm ; 9+0 flagellum ; motility ; helicoidal bending ; reactivation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The sperm flagella of the eel, Anguilla anguilla, are capable of vigorous motion in spite of having an axoneme with reduced structure that lacks the outer dynein arms, radial spokes and spoke heads, the two central tubules and the central tubule projections that are all part of the standard “9+2” axoneme. These sperm progress forward rapidly as a result of the propagation of helicoidal waves distally along the flagellum. Their flagellar beat frequencies are high, 93 Hz at 21°C, and they roll at a frequency of about 19 Hz. Eel sperm could be demembranated with Nonidet P-40 and reactivated with MgATP2- in 0.22 M K acetate at pH 8.1. The reactivated motility closely resembles that of the live sperm, with a beat frequency of 69 Hz, but the demembranated flagella are unusually fragile, and commonly disintegrate by a combination of splitting, coiling, and sliding within a few minutes. Little reactivation is obtained if acetate is replaced by Cl- in the reactivating medium. The Michaelis constant for beat frequency (0.2 mM) is similar to that obtained for several “9+2” flagella. These sperm, however, appear to lack the mechanism by which Ca2+ regulates waveform. Our results indicate that eel sperm flagella, which at rest are straight, are induced to bend helicoidally by ATP, as the result of sliding between tubules that is blocked at both the base and tip of the organelle. The flagellar waveform consists of a series of planar bends separated by short regions of right-handed twist, which give it an overall left-handed helicoidal form.
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 5 (1985), S. 377-392 
    ISSN: 0886-1544
    Keywords: newt ; lung ; cilia ; beat frequency ; waveform ; models ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Highly coupled newt lung ciliated cell models were used to study the effects of MgATP concentration on ciliary beat frequency and waveform. Models were prepared from ciliated lung cells of the newt Taricha granulosa by trypsin dissociation of the epithelium, demembranation with Triton X-100, and reactivation with MgATP, as described previously [Weaver and Hard, 1985]. Beat frequencies were measured stroboscopically. Ciliary waveforms of reactivated models and intact mucociliary epithelial sheets were determined by single frame analysis of high-speed movies. Waveform parameters calculated included the durations of the effective and recovery strokes, the angular velocities of the ciliary base and tip, the position of the bend along the ciliary shaft during the recovery stroke, the velocity of recovery stroke bend propagation, and the ratio of the duration of recovery stroke bend propagation to the duration of the recovery stroke itself. We found that beat frequency varied biphasically in response to MgATP at 21°C, as shown previously for isolated, individual, newt lung axonemes. Apparent Fmax (maximum beat frequency) and Km values of 25 Hz and 0.14 mM, and 35 Hz and 0.47 mM, respectively, were obtained for each linear segment of the biphasic double reciprocal plot. Demembranation did not alter either ciliary waveform or the pattern of coordination. In this system, metachrony is antilaeoplectic and ciliary waveform appears to be regulated independent of beat frequency.
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  • 7
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 5 (1985), S. 17-29 
    ISSN: 0886-1544
    Keywords: cell motility ; membrane recycling ; immunofluorescence microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Mouse peritoneal macrophages subjected to gradients of activated mouse serum were found by immunofluorescence observations to have their Golgi apparatus and their microtubule-organizing center largely oriented in the direction of the gradient. By analogy with similar results obtained with motile fibroblasts, it is proposed that these two organelles are rapidly and coordinately reoriented inside the macrophages in order to direct the insertion of new membrane mass, via vesicles derived from the Golgi apparatus, into the leading edge of the cell. Consistent with the importance of such membrane insertion to cell migration, we found that the ionophore monensin, an inhibitor of Golgi functions, inhibited cell motility in the chemostactic gradient. It was further shown that several inhibitors of chemotaxis (monensin, cytochalasin D, cycloheximide) did not inhibit the reorientation of the Golgi apparatus/microtubule-organizing center in cells exposed to a chemotactic gradient, and that the reorientation required extracellular Ca+2.
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  • 8
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 5 (1985), S. 491-506 
    ISSN: 0886-1544
    Keywords: Somitogenesis ; neurulation ; alpha-actinin ; morphogenesis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A discrete stage in two different morphogenetic processes has been examined employing fluorescently labelled alpha-actinin as a probe to localize native alpha-actinin and antibodies to localize fibronectin and collagen type I. The stage of somitogenesis examined is the transition from the compact mesenchymal somitic mass to the epithelial somitic vesicle (ie, epithelialization of the somite). The stage of neurulation examined is the transition from the relatively flat neuroepithelium to the approximation of the neural folds. Before these morphogenetic movements begin, the neuroepithelium is sitting upon a basal lamina and interstitial collagen, and the somite is surrounded by a meshwork of interstitial collagen. During both of these processes, the cells become narrowed at their apices in the region of the tissue that is becoming concave, and alpha-actinin is localized in the apices. The localization of intracellular alpha-actinin and extracellular fibronectin, and the distribution of collagen, suggest that there is a coordinated appearance and distribution of these molecules that is temporally associated with these discrete morphogenetic events.
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  • 9
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 5 (1985), S. 195-208 
    ISSN: 0886-1544
    Keywords: central pair ; radial spoke ; flagella ; mutant ; Chlamydomonas ; suppressor ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Flash photomicrography at frequencies up to 300 Hz and computer-assisted image analysis have been used to obtain parameters describing the flagellar bending patterns of mutants of Chlamydomonas reinhardtii. All strains contained the uni1 mutation, to facilitate photography. The radial spoke head deficient mutant pf17, and the central pair deficient mutant, pf15, in combination with suppressor mutations that restore motility without restoring the ultrastructural or biochemical deficiencies, both generate forward mode bending patterns with increased shear amplitude and decreased asymmetry relative to the “wild-type” uni1 flagella described previously. In the reverse beating mode, the suppressed pf17 mutants generate reverse bending patterns with large shear amplitudes. Reverse beating of the suppressed pf15 mutants is rare. There is a reciprocal relationship between increased shear amplitude and decreased beat frequency, so that the velocity of sliding between flagellar microtubules is not increased by an increase in shear amplitude. The suppressor mutations alone cause decreased frequency and sliding velocity in both forward and reverse mode beating, with little change in shear amplitude or symmetry.
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  • 10
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 5 (1985) 
    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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  • 11
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 5 (1985), S. 265-265 
    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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  • 12
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 5 (1985), S. 463-473 
    ISSN: 0886-1544
    Keywords: sponge dissociates ; cell migration ; time-lapse analysis ; cell aggregation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The production of both lamellipodia and peculiar thin protuberances (scleropodia) characterizes the preaggregative motility of cells after dissociation of the sponge Clathrina.The locomotory paths taken by cells before aggregation were recorded by time-lapse microcinematography. Changes of direction in successive 50-s time intervals and 50-s mean velocities of each cell were both taken into account as statistical variables. Their distributions give probability density curves that seem to fit bilateral exponential functions. The analysis of the angles of turn indicates a tendency for the cells to persist in their direction of motion and to make counter-clockwise turns. Implications of such in vitro cellular behaviors in aggregative and in vivo processes are suggested.
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  • 13
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 5 (1985), S. 81-101 
    ISSN: 0886-1544
    Keywords: fast axonal transport ; isolated axoplasm ; video microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The development of AVEC-DIC microscopy and the application of this method to the study of fast axonal transport in isolated axoplasm extruded from the giant axon of the squid Loligo pealei provides a new paradigm for analyzing the intracellular transport of membranous organelles. The size of the axon, the number of transported particles, and the absence of permeability barriers like the plasma membrane in this preparation permit many experiments that are difficult or impossible to perform using other model systems. The use and features of this preparation are described in detail and a number of properties are evaluated for the first time. The process of extrusion is characterized. Particle movement is evaluated both in the interior of extruded axoplasm and along individual fibrils that extend from the periphery of perfused axoplasm. The role of divalent cations, particularly Ca2+, and the effects of elevated Ca2+ on axoplasmic organization and transport are analyzed. A series of pharmacological agents and polypeptides that alter cytoskeletal organization are used to examine the role of microfilaments and microtubules in fast transport. Finally, the effects of depleting ATP and of adding ATP analogues are discussed. The extruded axoplasm preparation is shown to be an invaluable model system for biochemical and pharmacological analyses of the molecular mechanisms of intracellular transport.
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  • 14
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 5 (1985), S. 225-237 
    ISSN: 0886-1544
    Keywords: neural crest ; migratory behavior ; microfilaments ; stress fibers ; tractional force ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: We have investigated one aspect of the migratory behavior of quail neural crest (NC) cells by comparing the organization of microfilament bundles and the ability to distort migratory substrata by NC, somite, and notochord cells in vitro. In contrast to the numerous cytoplasmic stress fibers in somite-derived fibroblasts and notochord cells revealed by rhodamine-phalloidin staining and thin-section electron microscopy, microfilaments in NC cells are restricted to the cell cortex. To test the relative degrees of tension generated by these cell types on the underlying substratum, cells were cultured in collagen gels and on distortable silicone rubber sheets. Explanted somites and notochords produced dramatic radial alignment of 750 μg/ml collagen gels, whereas neural crest cells only aligned gels of lower concentrations. Fibroblasts did not migrate individually from explanted somites and notochords into 250 μg/ml collagen gels as readily as into higher concentration collagen lattices. In contrast, neural crest cells migrated into matrices of low concentration as well as into higher concentration collagen gels. Neural crest cells and their pigmented derivatives did not distort silicone rubber sheets, whereas somite and notochord-derived fibroblasts wrinkle this substratum after 4 days in culture. Thus, the differences in organization of the actin cytoskeleton reflect the tractional force exerted by these cells on their substratum. We hypothesize that the migratory behavior of NC cells in vivo may be related to their ability to translocate through embryonic extracellular matrices while generating relatively weak adhesions with the substratum, whereas the stronger forces generated by other embryonic cell types upon the delicate extracellular matrix may restrict their migration and may be associated with other morphogenetic events.
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  • 15
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 5 (1985) 
    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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  • 16
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 5 (1985), S. 31-51 
    ISSN: 0886-1544
    Keywords: microtubules ; birefringence ; flow birefringence ; tubulin ; polarization microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Understanding the molecular basis of mitotic movements in living cells will require correlative experiments on intact cells, cell models, purified tubulin, and perhaps other biopolymers. Birefringence is one assay that is useful in all of these experimental situations. Heretofore, studies of birefringence changes during mitosis have lacked a quantitiative basis for interpretation in terms of microtubule number and packing density. One of the aims of this work was to establish that relationship.Purified calf brain tubulin was polymerized to equilibrium and oriented in the hydrodynamic field of a microcapillary flow birefringence apparatus. The relationship between birefringence and microtubule packing density was determined by a combination of optical, electron microscopic, and biochemical methods. The data correlate surprisingly well with those obtained by others from in vitro measurements on isolated mitotic spindles. Using the flow birefringence data, the sensitivity of polarizing microscopes for detecting microtubules was examined and found to depend on microtubule packing density, object thickness, and instrumental factors that limit both the detection and measurement of weakly birefringent objects. Because of the dependence of measurement sensitivity on object thickness, a method of measuring the thickness of microtubule bundles using the dispersion of birefringence was developed. This method is capable of measuring thickness to within two or three Airy diffraction units and does not require any assumptions regarding object symmetry.
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  • 17
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    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 5 (1985), S. i 
    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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  • 18
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 5 (1985), S. 209-224 
    ISSN: 0886-1544
    Keywords: flagellar dynein ; cyclic nucleotides ; sperm activation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Methods of demembranation and reactivation of Lytechinus pictus sperm were developed that result in non-motile sperm which take on a stable bend of about 3.5 radians at the proximal end of the cell. The middle and distal portion of the flagellum is relatively straight or slightly bent in the same direction forming a somewhat “C” shaped sperm cell. In these studies, we refer to this characteristic shape as the quiescent form, and as opposed to “rigor wave” sperm, the quiescent form is induced and maintained by a relatively high concentration of MgATP2- (〉 0.2 mM). Other conditions important to the production and maintenance of the quiescent form in demembranated sperm include: starting with concentrated, undiluted sperm, maintaining low Ca++ in the demembranation buffer, using a minimum of 0.2 mM MgATP2- and pH of 7.9-8.1 in the reactivation buffer. Deviation from some of these conditions results in a dramatic increase in motile, asymmetrically beating sperm. Addition of 0.4 mM CaCl2 to the reactivation buffer increased the proximal bend angle to 5 radians. The induction and maintenance of the stationary bend is mediated by dynein activity: “rigor wave” sperm were transformed to the quiescent form upon 0.2 mM ATP addition; micromolar vanadate abolished the quiescent form by “relaxation” of the proximal bend; and the vanadate relaxed sperm were restored to quiescent form by catechol. Importantly, 20 μM cAMP activated motility of the otherwise quiescent-form sperm. Quiescent-form, demembranated sperm were also activated by mild trypsin digestion. These and other data suggest that the quiescent-form sperm are trapped at the end of the principal bend, and these data are consistent with the proposal that the single stationary bend results from asymmetry of active microtubule sliding [Gibbons and Gibbons, (1980): J. Cell Biol. 84:13-27].
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  • 19
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 5 (1985), S. 267-292 
    ISSN: 0886-1544
    Keywords: mammalian cilia ; respiratory tract ; mucociliary clearance ; laterofrontal cirri ; Mytilus edulis ; beat cycle ; computer analysis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The beat cycles of rabbit tracheal cilia in culture and Mytilus laterofrontal cirri were recorded using a phototransistor, transillumination, video, and phase-contrast microscopy. The photoelectronic signal and video image of the ciliary activity were simultaneously recorded as a composite image. The photoelectronic signal was converted into a digital signal by a data acquisition system for further computer processing.By the selection of a small detector area and accurate detector alignment, a simple, repetitive photoelectronic signal representing ciliary activity was obtained. This signal records the ciliary beat frequency and demonstrates the triphasic nature of the beat cycle. The photoelectronic signal can be precisely correlated with the ciliary activity by analysis of the composite video recordings to provide the duration of the effective, recovery, and rest phases of the beat cycle. The videophotoelectronic signal correlations were verified by high-speed cinematography. High-speed films of ciliary activity were digitized, and the image density of selected pixels was analyzed by computer with respect to time and ciliary motion.These studies indicate that duration of the phages of the beat cycle are differentially reduced with increased beat frequency; the effective phase duration was quickly reduced to a minimum. This was followed by the reduction of the duration of the recovery phase to a minimum. The rest phase continues to be reduced without reaching a minimum, over the range of beat frequencies observed. These results suggest that ciliary beat frequency may be regulated either by modifying the rates of dynein-microtubule interactions or the rate of transition from one beat phase to the next.
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  • 20
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    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 5 (1985) 
    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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  • 21
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    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 5 (1985), S. 351-354 
    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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  • 22
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 5 (1985) 
    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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  • 23
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 5 (1985), S. 447-461 
    ISSN: 0886-1544
    Keywords: chemokinesis ; orthokinesis ; klinokinesis ; polymorphonuclear leucocytes ; locomotion ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Evidence is presented to show that klinokinesis, which was previously demonstrated in bacteria and amoeba only, may also occur in metazoan cells. The chemotactic peptide formyl-Met-Leu-Phe (fMLP) elicited orthokinetic and klinokinetic responses of human blood-borne polymorphonuclear leucocytes (PMNs) under the test conditions used. Increased speed (orthokinesis) was due to an increase in the proportion of migrating cells as well as in the speed of the locomoting subset. The klinokinetic effect was manifested by a decrease in the klinolocomotion index, the mean angle of changes in direction ≥ 90°, and the frequency of turns ≥ 90°. The klinolocomotion index was inversely related to speed. This explains the synergistic effect of klinokinesis and orthokinesis in this system. Colchicine alone had and orthokinetic effect which was exclusively due to alterations in the proportion of migrating cells and it altered the turning behaviour without exerting a klinokinetic effect. However, colchicine had marginal orthokinetic and klinokinetic effects on fMLP-stimulated cells resulting in reduced translocation. The relationship between klinokinesis and mean angle or frequency of turns has been analysed. Klinokinesis was a substantial though not the major element of the chemokinetic response to fMLP under the conditions used. No other metazoan cells have been shown to possess such a complete pattern of responses, including orthokinesis, klinokinesis, and chemotaxis, which regulate locomotion.
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  • 24
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    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 5 (1985), S. 529-543 
    ISSN: 0886-1544
    Keywords: actin ; regulatory protein ; cytoskeleton ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: We have isolated a 30,000-dalton protein from Dictyostelium which cosedimented with and affected the low shear viscosity of actin. At low concentrations, this protein increased the low shear viscosity to greater than that of the actin control, whereas higher concentrations decreased viscosity. The viscosity decrease correlated with the formation of actin filament bundles, as seen electron microscopically. This protein resembled a previously reported actin-binding protein from Dictyostelium [Fechheimer and Taylor, 84, J Biol Chem 259:4514] in electrophoretic mobility, Stokes radius, and ability to crosslink filaments, but was shown to be different by peptide mapping, lack of immunologic crossreactivity, and lack of sensitivity to calcium.
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  • 25
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    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 5 (1985), S. 507-527 
    ISSN: 0886-1544
    Keywords: axonal transport ; microtubules ; organelles ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A model for fast axonal transport is developed in which the essential features are that organelles may interact with mechanochemical cross-bridges that in turn interact with microtubules, forming an organelle-engine-microtubule complex which is transported along the microtubules. Computer analysis of the equations derived to describe such a system show that most of the experimental observations on fast axonal transport can be simulated by the model, indicating that the model is useful for the interpretation and design of experiments aimed at clarifying the mechanism of fast axonal transport.
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  • 26
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    Cell Motility and the Cytoskeleton 5 (1985), S. 137-173 
    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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  • 27
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    Cell Motility and the Cytoskeleton 5 (1985), S. 475-489 
    ISSN: 0886-1544
    Keywords: digital image processing ; flagella ; cilia ; bends ; Hemicentrotus ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A novel method of digital image analysis of the bends of eukaryotic flagella and cilia was devised. In the analysis system, all image pixels were systematically extracted and processed to measure angular direction and curvature. Simulation experiments on theoretical model pictures of flagella with sine-generated or arcstraight line bending waves demonstrated that the method can be used with considerable high accuracy. This method then revealed abrupt changes in slope of the curvature in sperm flagella and embryo cilia of the sea urchin, Hemicentrotus pulcherrimus. This indicates that the digital image processing used may be helpful in the study of flagellar and ciliary movements.
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  • 28
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    Cell Motility and the Cytoskeleton 5 (1985), S. 61-75 
    ISSN: 0886-1544
    Keywords: dynein ; erythro-9-[3-2-(hydroxynonyl)]adenine (EHNA) ; ATPase ; inhibition ; axoneme ; cytoplasm ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: In current purification strategies, affinity for microtubules or calmodulin is used to identify and purify cytoplasmic dynein-like ATPase from cell-free extracts of unfertilized sea urchin eggs. However, affinity purification procedures, though they define dynein-like ATPase activity, have not yet proven to be quantitative. An alternative purification strategy capable of producing a high yield of enzyme would require a specific assay in order to monitor cytoplasmic dynein purity at each step.In this study, we make a detailed comparison of the effects of EHNA on 22 different ATP-metabolizing enzyme activities, including 13 Mg++-ATPases. We isolate cytoplasmic dynein-like ATPase activity from three species of sea urchin eggs and sperm and show by means of dose-response curves that their sensitivities to inhibition by EHNA are very similar to one another. We demonstrate further that the EHNA dose-response characteristics of fourteen other ATP-metabolizing enzyme activities, including seven nondynein Mg++-ATPases, differ quantitatively from those of dynein-like ATPases.In studies of three other agents (vanadate, Ca++/calmodulin, and Triton X-100), we find that dynein-like ATPases vary by two orders of magnitude in their sensitivities to inhibition by vanadate, and little or no stimulation by either Ca++/calmodulin or Triton X-100 is seen. Our results suggest that inhibition by EHNA is a universal and specific property of dynein-like ATPases, which ultimately should prove useful in the quantitative purification and characterization of cytoplasmic dynein-like ATPase (s).
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  • 29
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    Cell Motility and the Cytoskeleton 5 (1985), S. 103-122 
    ISSN: 0886-1544
    Keywords: gliding ; cell motility ; cytoskeleton ; diatoms ; capping ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Gliding motility was investigated in the marine diatom, Amphora coffeaeformis. Ultrastructural, biochemical, and pharmacological protocols were employed to probe the possible involvement of cytoskeletal proteins and a secretory process in gliding motility. Motility rate was measured using a video recording apparatus, and the effects of various cytoskeleton-disrupting drugs on motility were tested. Cytochalasins D and E, podophyllotoxin, and vinblastine (all at 25 μUg/ml) reversibly inhibited motility, as did monensin (10 μUM) and pronase (25 μUg/ml). Biochemical protein analysis of whole-cell extracts by one- and two-dimensional polyacrylamide gel electrophoresis revealed polypeptides comigrating with rabbit skeletal muscle actin and bovine brain tubulin; however, specific assays used to separate actin from whole-cell preparations gave ambiguous results. Ultrastructural studies revealed the presence of extracellular material between the raphe canal and the substratum in motile cultures. An assay was devised for the detection of radioactively labeled material (MW 〉 1800 Daltons) released by motile cultures into the culture medium. When cultures were treated with either an anticytoskeletal drug or monensin, motility was inhibited while the amount of measured radioactivity increased over solvent-treated control groups. The results from this study indicate possible roles for both actin- and tubulin-based structures in gliding motility of Amphora. Though secretion may be necessary for gliding to occur, its exact relationship to motility was not deduced. The data obtained in this study are compatible with a theory for the mechanism of gliding which involves the surface translocation of externally exposed membrane proteins against an immobile matrix of substratum-attached secreted material to generate the force required for movement.
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  • 30
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    Cell Motility and the Cytoskeleton 5 (1985), S. 123-136 
    ISSN: 0886-1544
    Keywords: locomotion and shape control ; epithelial cells ; calcium ; reflection-interference contrast-microscopy ; cinematography ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The role of calcium in the induction of locomotion, control of direction of locomotion, and modulation of shape of epithelial cells derived from Xenopus laevis tadpole epidermis is investigated. Local influx of calcium is achieved by electrophoretic release of small amounts of calcium from a micropipette (tip diameter 0.1-0.5 μUm) closely apposed to the cell body or lamella. The cells are made permeable for calcium by calcium ionophore A23187, and they are kept in Ca++-free, Mg++-rich EGTA Ringer. Another method used to induce Ca++ influx is local application of A23187 while cells move in normal culture medium.Influx of Ca++ into the lamella induces a localised increase in thickness and enlargement of the lamella. Stationary cells become active and show movement in the direction of the Ca++ gradient. Fried-egg-shaped cells tend to acquire a semicircular shape and start moving. Moving cells change the direction of their locomotion, following the direction of Ca++ release. Influx of Ca++ in the cell body region induces its contraction concomitant with an increase in lamellar area.These observations suggest the presence of two different Ca++-sensitive components: an actomyosin meshwork in the cell body and an actin gel in the lamella. Influx of Ca++ induces contraction of actomyosin and solation of actin gel. Interaction of these two systems would explain modulation of shape and generation of locomotion in epithelial cells.
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  • 31
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    Cell Motility and the Cytoskeleton 5 (1985) 
    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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  • 32
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    Cell Motility and the Cytoskeleton 5 (1985), S. 293-309 
    ISSN: 0886-1544
    Keywords: Mytilus edulis ; 5-hydroxytryptamine ; lateral cilia ; laterofrontal cirri ; beat frequency ; methylxanthine ; filter-feeding ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The laterofrontal (LF) cirri on isolated gill filaments of Mytilus edulis, prepared in natural seawater, are active and initially beat with an average frequency of about 8 Hz (with a range of 6-14 Hz). However, the lateral (L) cilia on these filaments are arrested in a position at the end of their recovery stroke. Perfusion of the filament with artificial seawater (ASW), with or without 1% ethanol, has little or no biological effect on the activity of the LF cirri, although a transitory decrease in frequency often accompanies the perfusion process. The L cilia remain arrested during perfusion with ASW. The exposure of the gill to low levels of 5-hydroxytryptamine (5HT) (10-8 〈 5HT 〈 10-7 M) has no effect on the activity of the LF cirri but stimulates the L cilia to beat. Exposure to higher concentrations of 5HT (〉 10-7 M) elevates the beat frequency of the L cilia and simultaneously inhibits the activity of the LF cirri, leading to their arrest in a position at the end of the effective stroke. This arrest of the LF cirri occurs as the L cilia attain a 5HT-induced beat frequency between 12 to 14 Hz. The influence of 5HT on the L cilia and the LF cirri can be reversibly mimicked or enhanced by the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX). A concentration of 0.5 mM IBMX mimics low 5HT concentrations (about 10-7 M) by stimulating the L cilia to beat without affecting the beat frequency of the LF cirri. A combimation of 10-7 M 5HT and 0.5 mM IBMX in ASW mimics high (〉 10-6 M) 5HT concentrations by arresting the LF cirri and increasing the beat frequency of the L cilia. Under these conditions, the threshold of the LF cirri arrest response is again found to occur as the L cilia attain a beat frequency of 12 - 14Hz. These results suggest that the mechanisms of LF cirri arrest and L cilia activation are mediated by 5HT -induced changes in intracellular cyclic AMP levels.
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  • 33
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    Cell Motility and the Cytoskeleton 5 (1985), S. 393-413 
    ISSN: 0886-1544
    Keywords: chromosome movement ; Colcemid ; nocodazole ; meiotic prophase ; microtubules ; vinblastine ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The effects of Colcemid, nocodazole, and vinblastine on microtubules and on the movement of chromosomes during late diakinesis were investigated in spermatocytes of the crane fly Nephrotoma suturalis. The kinds of movements observed in untreated cells - sex bivalent rotations, sex bivalent excursions, and rotations and positional changes of autosomal bivalents - also were observed in drug-treated cells. These results were obtained in living cells in which the disruption and inhibition of microtubule assembly had been confirmed with polarized light microscopy. Effects of Colcemid and nocodazole also were assessed in fixed cells using electron microscopy. The results are in agreement with a hypothesis that microtubules are not a force-generating component of the molecular machinery that brings about prophase movements.
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  • 34
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    Cell Motility and the Cytoskeleton 5 (1985), S. 53-60 
    ISSN: 0886-1544
    Keywords: calcium ; Chlamydomonas ; flagella ; motility ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Ca2+ has profound effects on the movement of cilia and eukaryotic flagella, including those of Chlamydomonas. Two clear changes seen in Chlamydomonas flagella with changes in Ca2+ are beat frequency and symmetry. Photographic and computer assisted analysis of flagellar bending patterns on a uniflagellate mutant of Chlamydomonas have been used to examine details of the effects of Ca2+ on the movement of ATP-reactivated, demembranated flagella. In addition to the forward mode bending pattern seen at low Ca2+ concentrations (10-9 M), which has a frequency of about 50 Hz and the reverse mode bending pattern seen at high Ca2+ concentrations (10-4 M) with a frequency around 70 Hz, we carefully examined bending patterns in the intermediate Ca2+ concentration range of 1-6.5 × 10-6 M. In this intermediate range, the bending patterns have significantly reduced asymmetry and slightly increased frequency, compared to the motility observed at low Ca2+ concentrations. These observations indicate that changes in these two parameters of motion do not occur in parallel and suggest that the effects of Ca2+ may be a multicomponent process. Physiologically, these changes in the beat pattern at intermediate Ca2+ may signal either (1) the beginning stages of transition to the symmetrical, high-frequency beating seen at high Ca2+, or (2) a more normal forward mode motility for the trans flagellum as suggested by Kamiya and Witman [1984]. No large amplitude bending patterns associated with transitions between forward and reverse mode beating in intact cells were seen at the intermediate Ca2+ concentrations.
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  • 35
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    Cell Motility and the Cytoskeleton 5 (1985), S. 415-430 
    ISSN: 0886-1544
    Keywords: Actin ; immunofluorescence ; NBD-phallacidin ; Chlamydomonas ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: We have localized actin in gametes of Chlamydomonas reinhardi by two approaches: (1) indirect immunofluorescence with an affinity-purified antibody and (2) staining with NBD-phallacidin, a fluorescent reagent that binds only to F-actin [Barak et al, 1980, Proc Natl Acad Sci, 77:980-984]. Staining of either mating type “plus” (mt+) or “minus” (mt-) gametes with antiactin antibody resulted in similar fluorescent images: most of the actin was located peripherally along the lateral and posterior aspects of the cells. There was diffuse staining centrally, but the flagella did not stain. No brightly stained spot was observed near the mt+ mating structure, the site where the fertilization tubule elongates with concomitant polymerization of actin [Detmers et al, 1983, J Cell Biol, 97:522-532]. Gametes stained prior to mating with NBD-phallacidin showed no fluorescence above background, indicating that there were no concentrations of F-actin in these cells. This suggested that the cytoplasmic staining observed with antiactin represented primarily a nonfilamentous form of the protein. In mating gametes staining with NBD-phallacidin was detected only in the fertilization tubule, indicating that this was the only dense accumulation of filamentous actin within the cells. Mating gametes stained with antiactin antibody exhibited cytoplasmic fluorescence that was slightly more punctate than prior to mating, and the fertilization tubule was brightly stained. Our observations suggest that the site-specific polymerization of actin within the fertilization tubule occurs in the absence of a concentrated pool of actin subjacent to the mating structure.
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  • 36
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    Cell Motility and the Cytoskeleton 5 (1985), S. 545-557 
    ISSN: 0886-1544
    Keywords: neutrophils ; cytoskeleton ; actin polymerization ; NBDphallacidin ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The studies presented here characterize a simple, quantitative NBDphallacidin extraction assay for determining the F-actin content of fMLP-activated neutrophils. The NBDphallacidin extraction assay is based upon the specificity of NBDphallacidin binding to F-actin and the solubility of NBDphallacidin in methanol. Cells are fixed, permeabilized, and stained with NBDphallacidin; the cells are then pelleted, the bound NBDphallacidin is extracted into methanol, and the RFI (excite 465; emit 535) of the solution is determined. Binding of NBDphallacidin to neutrophils is saturable and 90% of bound NBDphallacidin is displaced by nonfluorescent phalloidin. The extraction of bound NBDphallacidin into methanol is complete and the excitation/emission characteristics of NBDphallacidin are not altered by extraction. The assay is relatively inexpensive, applicable to the study of cells in suspension or on substratum, allows kinetic studies with 5-10s time resolution, and is not affected by the shape of the cell or the distribution of the probe. We used the NBDphallacidin extraction assay to study the kinetics of fMLP-induced change in the F-actin content of neutrophils and the effect of tBOC peptide, an inhibitor of fMLP binding, on these changes. The extraction assay reveals a rapid, sequential fMLP-induced increase followed by a decrease in F-actin content. The tBOC peptide inhibits fMLP-induced actin polymerization. Addition of tBOC during fMLP-induced polymerization or at times when F-actin content is maximal enhances F-actin depolymerization. The rate of F-actin depolymerization is ≥ fourfold faster in the presence than in the absence of tBOC. The results show that (1) The NBDphallacidin extraction assay is useful for studying the kinetics of change in F-actin content of nonmuscle cells; (2) fMLP receptor occupancy is required for fMLP-dependent polymerization but not depolymerization; and (3) both the actin polymerizing and depolymerizing processes are active in the cell within 5 s after fMLP stimulation. Implications of these observations for understanding the observed increase and, then, decrease in F-actin content of fMLP-activated cells are discussed.
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  • 37
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    Cell Motility and the Cytoskeleton 5 (1985), S. 311-322 
    ISSN: 0886-1544
    Keywords: Spectrin ; TW 260/240 ; chicken intestinal brush border ; actin assembly ; actin filament cross-linking ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: TW 260/240 is a tissue-specific spectrin found in the terminal web region of the chicken intestinal bruish border. We have examined the effects of TW 260/240 on assembly rates and critical concentrations (Co's) for monomer addition at the barbed and pointed ends of the actin filament. For these studies, acrosomal processes (AP) from Limulus sperm were used as nuclei for actin assembly. Under conditions which favor the interaction of TW 260/240 for actin (20-75 mM KCl, 2 mM Mg++) no effect on either elongation rates or Co's at either end of the actin filament was observed in the presence of this spectrinlike protein. The Limulus AP nucleation assay also allowed visualization of the kinetics of filament binding and cross-linking by TW 260/240. Ultrastructural analysis of TW 260/240 binding to actin filaments at their growing ends indicates that TW 260/240 tetramers bind laterally to the filament. Finally, evidence is presented that indicates that filaments cross-linked by TW 260/240 are stabilized against shear-dependent breakage.
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  • 38
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    Cell Motility and the Cytoskeleton 5 (1985), S. 323-331 
    ISSN: 0886-1544
    Keywords: Tetrahymena ; cell model ; ATP concentration ; Ca sensitivity ; backward swimming ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Using Tetrahymena pyriformis, strain w, and Tetrahymena thermophila, B-1868, we prepared cell models that showed ciliary reversal with change in Ca-ion concentration, as was also noted for the Paramecium cell model. No differences could be found between these two strains in the reactivation state, and their response to environmental conditions was essentially the same. The reactivation rate was 90% or more. Swimming velocity of the cell model was found to be 200 ± 49 μm/sec at 25.0°C ± 0.5°C, while the velocity for the living cells was 527 ± 101 μm/sec. Swimming velocity with change in environmental conditions, such as pH, Mg-ATP, and Ca-ion concentrations, was studied. Compared to the cell model of Paramecium, the Tetrahymena cell model had higher sensitivity toward Ca-ion in the reactivation medium. The effects of chlorpromazine, and inhibitor of calmodulin, on the swimming behavior of the cell models were studied.
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  • 39
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    Cell Motility and the Cytoskeleton 5 (1985), S. 431-446 
    ISSN: 0886-1544
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Microtubules assembled from sea urchin eggs with the use of taxol contain a 77,000-dalton protein as the major nontubulin component [Vallee and Bloom (1983): Proc Natl. Acad. Sci. U.S.A. 80:6259-6263]. We have raised five monoclonal antibodies to this protein to aid in its characterization. Immunoblot analysis of the sea urchin microtubule purification fractions indicated that the protein copurified quantitatively with microtubules. All five antibodies stained the mitotic spindle of dividing sea urchin eggs by immunofluorescence microscopy, indicating that the protein was a component of the mitotic spindle and suggesting that it was actually localized on microtubules in vivo. Immunofluorescent staining of higher resolution was observed in a subpopulation of the coelomic cells found in adult sea urchins, confirming that the 77,000-dalton protein is indeed present on microtubules in vivo. Because taxol was not used for the immunofluorescence experiments, we conclude that the microtubule-associated protein (MAP)-like behavior of the 77,000-dalton protein in vitro was not induced artifactually by taxol. To determine whether this protein is a component of sea urchin microtubules in general, cilia obtained from blastula stage embryos and sperm tail flagella were analyzed with the antibodies. The protein was undetectable by both immunoblot analysis and immunofluorescence microscopy in both preparations of axonemal microtubules. These results indicated that the 77,000-dalton MAP is restricted to cytoplasmic and mitotic microtubules in the sea urchin. Furthermore, in view of its particular abundance in embryos, whose microtubules are devoted substantially to mitosis, the 77,000-dalton MAP is likely to play an important role in regulating the activity of mitotic spindle microtubules in the sea urchin.
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  • 40
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    Cell Motility and the Cytoskeleton 5 (1985), S. 251-263 
    ISSN: 0886-1544
    Keywords: Chlamydomonas ; movement ; flagellar beat ; stigma ; high-speed microcinematography ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Using high-speed microcinematography (100-500 f/s) the movement of Chlamydomonas reinhardtii mutant 622 E has been studied with frame-by-frame analysis. The stigma lies in the cell equator, displaced out of the flagellar plane anticlockwise by an angle of about 45°. During forward movement the cells rotate anticlockwise about their long axis with a frequency of 1.4-2 Hz (maximum 2.5 Hz). The rotation is caused by a lateral component of 3-dimensional beating of the flagella during the effective strokes. The helical swimming path is a result of an unequal flagellar beating. This is normally synchronous, but synchrony is interrupted from time to time by an additional beat of the outward directed flagellum, in our study one after about every twenty beats on average. These results are discussed and compared with the results published by other groups.
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  • 41
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    Cell Motility and the Cytoskeleton 5 (1985), S. 355-375 
    ISSN: 0886-1544
    Keywords: Newt ; lung ; cilia ; cell models ; ciliary coordination ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Demembranated ciliated cell models are useful for studying mechanisms responsible for the regulation of ciliary coordination and waveform. This paper describes procedures for isolating ciliated cells from the newt, Taricha granulosa, by trypsin dissociation, their subsequent demembranation by Triton X-100, and their reactivation with MgATP to produce highly motile, coordinated, ciliated cell models. Reactivation of cell models with a high degree of mechanochemical coupling depended on avoiding mechanical damage and maintaining optimal conditions during all stages of isolation and reactivation. Highly motile models were prepared from cells incubated in trypsin, treated briefly with EDTA, separated by gentle agitation, and concentrated by centrifugation at low gravitational forces. Optimal demembranation and reactivation conditions were similar to those described previously for isolated newt lung axonemes. Under these conditions, nearly 100% of the models were reactivated when provided with MgATP and 90-95% beat with coordinated waves. The ciliary tufts beat at frequencies within the range measured in living cells and their reactivated motility was stable for at least 30 min at constant MgATP. These highly coupled models were used to show (1) that development of coordination in the ciliary tuft occurs at a higher substrate concentration range (10-25 μM) than that required to initiate motility per se (2-10 μM); (2) that outer dynein arms may not contribute to beat frequency at substrate concentrations below 35 μM; and (3) that vanadate has effects both on beat frequency and coordination of the tufts.
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    The @Anatomical Record 211 (1985) 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
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    The @Anatomical Record 211 (1985), S. 1-8 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Ultrastructural localization of carbonic anhydrase was determined by applying Hansson's histochemical method to glutaraldehyde-fixed frozen sections of guinea pig peritoneal polymorphonuclear leukocytes (PMNs) and lysosomes isolated from rat liver tissue after the animal had been injected with Triton WR-1339. A positive histochemical reaction for carbonic anhydrase in PMNs was found in the matrix of lysosomes. After PMNs phagocytized polystyrene latex particles or emulsified paraffin oil droplets, a positive reactivity for carbonic anhydrase was found in the space between the lysosomal membrane and the particle. Liver lysosomes also revealed positive carbonic anhydrase histochemical reactivity. To confirm the histochemical reaction, indirect immunoferritin labeling was conducted with rabbit antibody to human red blood cell carbonic anhydrase C on glutaraldehyde-fixed, freezethawed human PMNs. Immunolabeling was observed in lysosomes. These results suggest that carbonic anhydrase is a constituent of lysosomes of PMNs and liver cells.
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    The @Anatomical Record 211 (1985), S. 17-23 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The effect of adriamycin on rat incisor was investigated 1 day after administration. Rats were injected intravenously with adriamycin at a dose of 5 mg/kg body weight and sacrificed by perfusion with a 2.5% phosphate buffered glutaraldehyde solution. The principal effect of the drug on the incisor was the production of cell degeneration. This was extensive in the apical region, being present along the entire periphery of the dental papilla. In sections examined more incisally, cell degeneration gradually disappeared from the labial portion of the incisor but was present in the lingual portion. This degeneration of cells was not present at the site where mature odontoblasts had differentiated on the lingual surface of the pulp chamber. It appears that the affected cells were early preodontoblasts and the precursors of preodontoblasts. However, as these cells became more differentiated they apparently became more resistant to the drug's effect.
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    The @Anatomical Record 211 (1985), S. 24-28 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The external genitalia of four adult female pygmy chimpanzees (Pan paniscus) were examined during a 2-year period. It was found that the labia majora are retained in adults of this species and that, when tumescent, the labia minora effectively relocate the frenulum and clitoris so that they point anteriorly between the thighs. When detumescent, the configuration of the labia minora and clitoris resembles that of immature common chimpanzees (P. troglodytes). It is suggested that the simple, structural relocation of the clitoris from the normal condition noted in adult P. troglodytes makes possible the homosexual, intergenital rubbing observed in P. paniscus, when ventroventral juxtaposition of the individuals permits eye-to-eye contact. In addition, this change probably increases sexual stimulation of the female during heterosexual, ventroventral copulations. The possibility of intense, face-to-face sexual interactions has been achieved by P. paniscus without the evolution of bipedalism and the consequent reorientation of the genitalia seen in Homo sapiens.
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    The @Anatomical Record 211 (1985), S. 9-16 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The ability of PGE2 to stimulate bone resorption in vitro and in vivo is well established but the effects of this compound on bone formation are still controversial. Recent clinical reports have suggested that long-term infusion of PGE in infants with cyanotic heart diseases led to a stimulation of periosteal bone formation and to hyperostosis.In the present report, we describe the effects of PGE2 (10-5 M) in bone organ cultures on bone resorption, measured by the release of 45Calcium and the number of osteoclasts in sections of cultured bones, and bone volume, by measuring separately medullary and cortical areas. PGE2 induced a marked increase in 45Ca release and in cortical and medullary osteoclast numbers over 4 days in vitro; despite this increase in bone resorption, cortical bone volume remained constant, indicating a parallel increase in bone resorption and formation at this site. Morphological and quantitative data demonstrated a higher extent of osteoblastic surface along the periosteum of PGE2-treated bones when compared with control cultures. Medullary bone volume, on the other hand, decreased sharply during the culture period, demonstrating a lack of parallel increase in bone formation at this site.It is concluded that, under these experimental conditions, prostaglandin E2 stimulated both resorption and formation along the periosteum and only bone resorption along the endosteum of the cultured bones. The overall effect of PGE2 on bone as a whole, however, was net bone loss.
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  • 47
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    The @Anatomical Record 211 (1985), S. 29-33 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Since previous studies have shown that an active pineal gland exerts an inhibitory effect on circulating levels of thyroxin in the Syrian hamster, a study was conducted to determine whether the histology and ultrastructure of the thyroid gland supported the conclusions drawn from the hormone data. The ultrastructure of thyroid glands of blinded male Syrian hamsters was compared to that of intact controls kept under a 14L/10D photoperiod, to that of blinded hamsters also pinealectomized, and to that of blinded hamsters receiving 80 μg/ml of melatonin in the drinking water. Serum thyroxin (T4) and serum thyrotropin (TSH) concentrations were determined by radioimmunoassay. After 10 weeks serum thyroxin concentrations were less than 50% of controls and concentrations were significantly reduced. EM examination revealed that blinded hamsters had an increased number of follicular cells with flattened epithelium and nondilated endoplasmic reticulum compared to intact controls. In blinded hamsters that were pinealectomized or treated with melatonin in the drinking water, the ultrastructure of the thyroid was not different from controls and serum thyroxin concentrations were restored to near normal. These ultrastructural data support the conclusion that the pineal gland is required to obtain inhibition of the pituitary-thyroid axis in blinded hamsters and that melatonin has a counter-inhibitory effect when administered via the drinking water.
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  • 48
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    The @Anatomical Record 211 (1985), S. 43-47 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The distribution of the intermediate filament protein vimentin in peripheral lymphoid tissues was determined using a monoclonal antibody. Frozen sections of tissue were stained using an avidin-biotin immunoperoxidase method. The antibody stained endothelial cells in spleen, lymph node, and tonsil. Unusual rod-like structures were revealed in the sinusoid-lining cells of the spleen. A variety of reticulum cells was detected, including fibroblastic reticulum cells, histiocytic reticulum cells (tingible body macrophages), and splenic marginal zone macrophages. Very few lymphocytes were immunoreactive. Only weak cytoplasmic immunoreactivity was observed in lymphocytes of the periarteriolar lymphocyte sheath of the spleen. The monoclonal antibody employed appears to be of limited usefulness in detecting normal lymphocytes, but is strongly reactive with endothelial structures and some types of reticulum cells.
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  • 49
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    The @Anatomical Record 211 (1985), S. 48-56 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Mouse blastocysts were cultured in hanging drops for up to 6 days in order to study development under conditions that avoid the distortion of embryos typically seen when they are allowed to attach to a glass or plastic surface. The survival rate of embryos in hanging drops was equal to that of embryos attached to culture dishes and superior to that of embryos suspended in gyrating flasks. Development of the embryonic portion was similar to that in vivo and on culture dishes but slower than in vivo; the egg cylinder stage was reached after 8-10 equivalent gestation days (4 to 6 days in culture), while that stage is reached at 5.5 to 6 days in vivo. The trophectoderm, however, developed in a unique manner. The cells migrated away from the inner cell mass (ICM), similar to embryos on a culture dish, but without a surface on which to spread they clustered distal to the ICM. In vivo, trophectoderm remained covering the ICM. By 5 days in hanging drop culture the embryos had developed a segmented appearance with trophoblast giant cells at the abembryonic pole, extraembryonic cells not covered by vacuolated endoderm in the central region, and embryonic endoderm surrounding a developing proamniotic cavity in embryonic ectoderm at the embryonic pole. These observations suggest that the trophectoderm is able to follow a developmental program independent of that in the embryonic portion and that its behavior is dominated by the different adhesive properties of the trophoblastic and embryonic cells.
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  • 50
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    The @Anatomical Record 211 (1985), S. 57-68 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Cephalic neural crest (NC) cells enter a cell-free space (CFS) that contains an abundant extracellular matrix (ECM). Numerous in vitro investigations have shown that extracellular matrices can influence cellular activities including NC cell migration. However, little is known about the actual ECM composition of the CFS in vivo, how the components are distributed, or the nature of NC cell interactions with the CFS matrix. Using ultrastructural, autoradiographic, and histochemical techniques we analyzed the composition and spatial organization of the ECM found in the CFS and its interaction with mesencephalic NC cells. We have found that a specific distribution of glycoproteins and sulfated polyanions existed within the CFS prior to the translocation of NC cells and that this ECM was modified in areas occupied by NC. The interaction between the ECM components and the NC cells was not the same for all NC cells in the population. Subpopulations of the NC cell sheet became associated with ECM of the ectoderm (basal lamina) while other NC cells became associated with the ECM of the CFS. Trailing NC cells (NC cells that emerge after the initial appearance of NC cells) encountered a modified ECM due to extensive matrix modifications by the passage of the initial NC cell population.
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  • 51
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    The @Anatomical Record 211 (1985) 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
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  • 52
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    The @Anatomical Record 211 (1985), S. 102-109 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The aim of this study was to develop a set of practical metamorphic stages for the tiger salamander Ambystoma tigrinum that would (1) provide an early and quantifiable marker of the onset of metamorphosis and (2) not be dependent on knowledge of maximal values of gill length or tail height. Twenty morphological parameters were obtained from Ambystoma tigrinum larvae as they progressed through laboratory-induced metamorphosis (21°C, continuous light). These parameters included measures of body weight, gill length, and tail height. Initiation or duration of metamorphosis could not be predicted when parameters were expressed relative to the number of days larvae were held in laboratory conditions.Early phases of metamorphosis were characterized by regression of the lower tail fin. Selected morphological parameters were expressed relative to the day on which each subject exhibited complete regression of the lower fin. This technique made it possible to order these parameters into a progressive series of metamorphic phases. Seven stages of metamorphosis were derived from this series, ranging from larvae (stage I) to complete regression of the gills (stage VII). Application of these stages does not require prior knowledge of maximal (initial) values of gill length or tail height.
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  • 53
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    The @Anatomical Record 211 (1985), S. 126-132 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The purpose of this paper is to develop statistical methods that take radiation spread into account in analyzing data from different autoradiographic experiments. The method uses the probability circle analysis of Salpeter and McHenry (1973) to obtain the probable source of each radioactive emission and the circle and point counting method of Williams (1969) to estimate the relative area occupied by each cellular site. Two levels of analysis are presented. The first level of analysis is concerned with estimating relative activities and standard errors for cellular items that are larger than the probability circle. The second level of analysis involves estimating relative activities and standard errors for cellular sites that are smaller than the probability circle and are therefore observed in circles containing another item such as cytoplasmic matrix. Two different tests of hypotheses are discussed. The first null hypothesis is that the radioactivity is randomly distributed among the cellular sites. The second null hypothesis is that there is no difference between two different treatments in the relative activities for a given site.
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  • 54
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    The @Anatomical Record 211 (1985), S. 117-125 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The role of calcium in initiating smooth muscle contraction is widely accepted. The sources of this calcium are thought to be located both intracellularly and extracellularly. We have recently developed a method by which the cellular localization of calcium in smooth muscle can be determined. This method involves exposing the tissue to 45Ca, rapidly freezing and vacuum dehydrating the tissue, and preparing the tissue for electron microscopic autoradiography (EM ARG). In the present study the distribution of calcium in control and potassium-contracted tissue of the rabbit vas deferens was compared. No significant differences in distribution were observed in the two treatments. This finding provides morphological support for the hypothesis that the calcium used in potassium-induced contraction is primarily of extracellular origin. In addition, significant sequestration by intracellular organelles does not occur during a potassium contraction. In other experiments, the effect of rinsing tissue in cold calcium prior to freezing was investigated. From these data is appears that calcium is removed from the cytoplasmic matrix, plasma membrane, and organelles in a nonuniform manner. Further investigation into these findings should enable us to characterize more precisely the intracellular redistribution of calcium that occurs as a result of a variety of physiological manipulations.
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  • 55
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    The @Anatomical Record 211 (1985), S. 133-141 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: For periods ranging from 26 to 87 weeks, the morphological characteristics of the flexor carpi radialis (FCR) muscle were examined in four cats trained to perform weight-lifting exercise. Four untrained, sex and weight-matched cats served as controls. The right FCR from each cat was surgically isolated, attached to a tension transducer, and set at its optimal length. The forelimb was perfused with 2% glutaraldehyde in 0.1 M cacodylate buffer. Small bundles of fibers were teased from their origin and insertion tendons and embedded in Epon. Spaced serial sections were used to examine the morphological features of the fibers for trained and control animals. Ultrastructural examination revealed muscle fiber degenerative changes, such as pyknotic nuclei, disruption of the sarcolemma, vacuolation, and disorganization of myofilaments. Such changes were observed at a higher frequency in trained muscle than in control muscle. Spaced serial sections of fiber bundles showed that the degree of degeneration varied along the length of the fiber. Fiber area measurements showed that trained muscle had both larger and smaller fibers than control samples. The very small fibers observed in the trained muscle were considered to be regenerating or “new” fibers since they had not undergone degenerative changes. “Satellite-like” cells were observed in trained muscle. Such cells resembled satellite cells but also contained developing myofilaments. Since evidence of degeneration-regeneration was observed in control samples, but at a lower frequency, it was postulated that weight-lifting exercise accelerates muscle fiber turnover in the cat FCR.
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  • 56
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    The @Anatomical Record 212 (1985), S. 282-287 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Testes of sexually mature men were studied histochemically with 20 fluorescein isothiocyanate-labeled lectins. Based on their pattern of reactivity with intratesticular spermatogenic cells, lectins were divided into five groups: (1) lectins reacting with all spermatogenic cells (Suc. ConA, WGA, LCA, PHA-E, PHAL, STA, MPA, and RCA-II); (2) lectin reacting with spermatocytes, spermatids, and spermatozoa, but not with spermatogonia (RCA-I); (3) lectins reacting with spermatids and spermatozoa only (BPA, PNA, SBA, and VVA); (4) lectins reacting only with spermatozoa (HPA, GSA-I, UEA-II, and GSA-II); and (5) lectins with no distinct staining of spermatogenic cells (DBA, LBA, and UEA-I). All lectins from groups 1-4 were reactive with ejaculated spermatozoa. On the basis of the staining patterns of the head region of ejaculated spermatozoa, four lectin reactivity groups were defined: (1) lectins reacting with the plasma membrane of the whole head (BPA, WGA, LCA, STA, RCA-II, PHA-E, PHA-L, RCA-I, UEA-II, and GSA-II); (2) lectin reacting with the acrosomal cap and postacrosomal region of the plasma membrane (Suc. ConA); (3) lectin reacting with the acrosomal cap region of the plasma membrane (PNA); and (4) lectins reacting with the midregion of the sperm head in a bandlike manner (HPA, VVA, SBA, GSA-I, and MPA). These data provide a map of lectin binding sites on human testicular spermatogenic cells and ejaculated spermatozoa and show that the distribution of glycoconjugate domains of spermatogenic cell changes during differentiation and maturation.
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  • 57
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    The @Anatomical Record 212 (1985), S. 301-306 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Target cells for 1,25(OH)2 vitamin D3 metabolites are identified in developing rodent teeth by the use of thaw-mount autoradiography. Following the injection of [26, 27-3H]-1,25(OH)2 vitamin D3 into 18-day- and 20-day-old fetal rats and neonatal mice, nuclear concentration of radioactivity is found in different cell types. In incisors of both animal groups, strong nuclear labeling is present predominantly in pulp cells, while relatively weakly labeled cells are found in the layers of odontoblasts, ameloblasts, and stratum intermedium. In molars, nuclear labeling is absent in fetal rats, but is present in 2-day-old neonates in pulp cells and cells in the layers of stratum intermedium of the first molars, but not in the second molars. The absence of labeled pulp cells in the progenitor regions of incisors and in molars of 20-day-old fetal rats, and differential ontogenic appearance of labeled pulp cells in molars, indicates that there is a critical period of receptor emergence. The finding that labeled pulp cells exist in the regions of incisors and molars where secretory odontoblasts are present suggests that nuclear uptake of 1,25(OH)2 vitamin D3 is related to cell maturation and differentiation, and topographically related to the formation of dentin. The results further suggest that, in contrast to bone, the predominant effect of 1,25(OH)2 vitamin D3 is not on tooth cells which are directly involved in the formation of calcified tissue, i.e., ameloblasts and odontoblasts, but rather on supporting tissues such as pulp cells and stratum intermedium.
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  • 58
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    The @Anatomical Record 212 (1985), S. 326-326 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
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  • 59
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    The @Anatomical Record 212 (1985) 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
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  • 60
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    The @Anatomical Record 211 (1985), S. 414-419 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The effects of the diabetes (D) mutation on utero-ovarian structure and function were examined in match-paired D and control (C) C57BL/KsJ mice between 2 and 16 weeks of age. Between 4 and 8 weeks of age, the uterine epithelium of D mice exhibited a remarkable increase in the amount of cytoplasmic lipid stores as compared with that of C animals. Associated with progressive hyperglycemia between 8 and 16 weeks of age, uterine atrophy and continued lipid accumulation occurred. Both serum progesterone and estradiol levels were lower in D than C mice between 8 and 16 weeks of age. In addition, the uteri of D mice failed to accumulate as much 3H-estradiol as C uteri at 16 weeks of age. These data demonstrate that changes in uterine structure that are attributable to depressed ovarian activity in diabetics underlie the reproductive failure in these animals. The temporal association between the onset of reproductive tract involution and the expressed hyperglycemic condition suggests a causal association between these events in the genetically diabetic C57BL/KsJ mouse.
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  • 61
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    The @Anatomical Record 211 (1985), S. 420-426 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Measurements of whole population cell kinetics of mouse intestinal epithelium during postnatal development are reported. Swiss albino mice aged 1, 2, 3, 4, 6, 8, 10, 12, 16, 19, 24, and 28 weeks were studied. Isolated epithelial preparations of jejunum and colon were used. Most kinetic parameters studied either increased or decreased with age to reach a steady level sometime after weaning. For example, before weaning about 30% of crypts were observed to be branching, while after weaning, the population of crypts that were branching decreased to adult levels of 5-10% in jejunum and 1-2% in colon. Thus, there was very active crypt formation before weaning, which likely continued into adult life but at a lower level. Villus formation appeared to be occurring in animals before weaning (i.e., 1-3 weeks), while it stopped with weaning, and thus the mean villus height increased to a plateau, which was constant with age after 4 weeks. In contrast, the mean villus width increased steadily with age. As the width of villi increased with age, the number of crypts assoiated with a villus also increased (presumably as a result of net crypt production in the adult). These measurements and many others (proportion of cells in S phase, number of cells/cm2, number of cells/villus, number of cells/crypt, etc.) are described.
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  • 62
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    The @Anatomical Record 211 (1985), S. 184-191 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The lymph node contains blood vessels of a special type, termed “high endothelial venules” (HEVs), which are involved in the process of lymphocyte recirculation. In standard tissue sections, many HEVs exhibit a nearly closed or closed lumen containing small lymphocytes but few, if any, erythrocytes. The question arose as to whether the appearance of HEVs in tissue sections is influenced by the routine method of animal sacrifice and/or of tissue processing. Therefore, the present work investigated the effects on HEVs of sacrificing rats as well as of excising and fixing their nodes with various procedures. It was observed that procedures involving animal bleeding or blood loss from nodes increase the percentage of HEVs exhibiting a nearly closed or closed lumen. The results further revealed that the endothelial thickness and other morphological features of HEVs are modified by this artifactual narrowing of HEVs. The possible significance of the phenomenon is discussed.
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  • 63
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    The @Anatomical Record 211 (1985), S. 174-183 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: This study used membranous chorion of the macaque monkey placenta to examine uptake and processing of exogenous proteins. Tissue was incubated with either cationic or anionic horseradish peroxidase. Incubation time was varied between 5-25 min to follow the endocytic pathways. In spite of some differences in binding, uptake and processing of the isozymes was similar. In the presence of tracers at 37°C both horseradish peroxidases were taken up in large (150-175) nm diameter coated vesicles. In addition, coated tubules 300-400 nm in length and 50-100 nm in diameter were seen in the apical cytoplasm. Often, the tubules bud off small (85-105 nm diameter) protein-filled coated vesicles which traversed the cytoplasm and fused with the basal-lateral plasma membrane. In other cases, the tubules or vesicles lost their clathrin coats and fused to form larger endocytic vesicles which later fused with phagolysosomes. After longer incubation, larger uncoated vesicles (endosomes) were seen releasing their contents at the basal-lateral membrane. These results suggest that multiple transport pathways exist in this epithelium. The first, involving only coated structures, may function to sort and concentrate specific ligands important for embryonic development. The second, involving the formation and translocation of large uncoated vesicles to the basal-lateral membrane, may also provide nutrients to the embryo. A third pathway directs the protein to phagolysosomes where it is presumably degraded. Degradation products could be used by the cells of the membranous chorion or may provide nutrients to the embryo.
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  • 64
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    The @Anatomical Record 212 (1985), S. 353-357 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Hypophysectomized female hamsters primed with 30 i.u. pregnant mare serum gonadotropin (PMSG) were injected with anti-PMSG serum 72 h later to induce atresia in Graafian follicles. The earliest histological evidence of atresia was seen in the cumulus oophorus at 4 h with a significant increase in pyknotic nuclei. The mural granulosa cells, however, did not show pyknoses until 12 h following anti-PMSG serum. The highest number of pyknotic nuclei in the cumulus oophorus was seen at 24 h, when pseudomaturation of the oocyte occurred. Breakdown of the mural granulosa cells was greatest by 48 h. Morphometric measurements of the follicles showed a significant decline in the thickness and area by the granulosa layer at 48 h following injection of anti-PMSG serum.The thecal compartment showed a decline in number of mitoses at 4 h and 12 h and in area at 48 h. However, no pyknotic thecal cells were observed during the 72 h following injection of anti-PMSG serum. The results support previous work using this model, which showed major biochemical changes in the granulosa cells at 12 and 48 h. The theca, however, was comparatively unaffected during atresia. The various studies using this model shew changes in atretic follicles similar to those seen in other models and in the intact animal. The major advantage of this medel is that it produces a large synchronous population of follicles that rapidly becomes atretic.
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  • 65
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Adrenergic nerves reach the ovary via two routes: along the arteries to the ovary and via the suspensory ligament. Results from earlier investigations suggest that denervation of the nerves along the arteries does not influence the ovulatory process. In the present study we have examined whether denervation by transection of the ovarian suspensory ligament influences the ovulatory process.Partial denervation of the ovary by transection of the ovarian suspensory ligament, sham operation, or only anesthesia were performed on immature 25-day-old rats. To induce ovulation, pregnant mare serum gonadotropin (PMSG) was injected in the morning (0800-0930), when the rats were 26 days old. This PMSG treatment normally induces ovulation around 0200 in the early morning of day 29 with subsequent formation of corpora lutea. Rats were killed 5-8 hr, 3 days, and 5 days after this ovulation time.Ovarian interstitial norepinephrine levels were markedly decreased after transection of the suspensory ligament. Ovulations had occurred in all denervated, as well as sham-operated, and control rats. The various groups did not differ in the number of ovulations per rat. Thus, the adrenergic nerves in the suspensory ligament appear not to be necessary for ovulation. Whether catecholamines themselves play a role in the ovulatory process cannot be elucidated from this experiment, since the norepinephrine content in the ovary was not totally depleted. It seems unlikely that adrenergic nerves reach the corpus luteum via the suspensory ligament, since transection of this structure did not change the luteal content of norepinphrine.
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  • 66
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    The @Anatomical Record 213 (1985), S. 410-420 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Small-granule APUD cell clusters and their clear-cell precursors were mapped in serial 2-μm glycol methacrylate-embedded, periodic acid-Schiff(PAS)-lead hematoxylin-stained sections of 13-, 14-, and 15-day fetal hamster lungs. Every sixth section was drawn from a camera lucida projection on tracing paper. Each tracing included the profiles of nonalveolated air passages and the locations of smallgranule cell clusters and solitary clear cells. Airways containing ciliated cells and those surrounded by condensed mesoderm were also labeled. Single clear cells were rare in fetal hamster lung. Of 2,368 endocrine cell loci identified in the three fetal age groups examined, only 14 were single clear cells. A preliminary survey of the entire left and right lungs showed that the pattern of airway and small-granule cell development in the infracardiac lobe was similar to that occurring in the remainder of the lung; this lobe was accordingly considered a model for the whole lung, and the ontogeny of its small-granule cell population was quantitated and compared with results of similar quantitative mapping of this lobe in an adult animal (Hoyt et al., 1982a,b). Along the lobar bronchus of the 13-day infracardiac lobe and proximal portions of its main branches, small-granule cell clusters occurred most often near airway intersections. As the number and density increased in subsequent fetal stages, small-granule cell clusters became conspicuous along internodal bronchial segments. In distributing bronchioles, the population density of small-granule cell clusters decreased between 13 and 14 days but more than doubled by day 15. Unlike human lungs, where centrifugally developing small-granule cell clusters are firmly established in terminal bronchioles well before birth, most peripheral bronchioles in fetal hamster were devoid of small-granule cell clusters, even at 15 days, one day before birth. Comparison of numerical population densities in this lobe of fetal and adult lungs indicates that small-granule cell clusters continue to form past day 15 and suggests that they are considerably more numerous in adult than fetal lung.
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  • 67
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    The @Anatomical Record 213 (1985), S. 94-101 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The ketone body B-hydroxybutyrate (B-OHB) produces malformations and ultrastructural alterations in mitochondria of mouse embryos exposed for 24 hours to the compound in whole embryo culture. The present study was conducted to establish the time-course of the mitochondrial changes to determine whether the changes are reversible, and to relate these changes to the malformations produced by the compound. Since mitochondria also play a key role in the metabolism of ketone bodies, the capacity of the early somite embryo to metabolize B-OHB was investigated in an effort to link the morphological alterations in the mitochondria to a biochemical process. Early somite embryos were cultured 4, 8, or 24 hours in the presence of 32 mM DL-B-OHB and then cultured for an additional 24 hours in control serum. Finally, embryonic tissue during the teratogenic period was assessed for its capability to oxidize B-OHB using D-(3-14C)-B-OHB. The treated embryos showed progressive alterations in the mitochondria, beginning at 4 hours with a loss of matrix density and culminating at 24 hours with high-amplitude swelling, complete loss of matrix density, and disappearance of cristae. These alterations were reversible following removal of the embryos after 24 hours of exposure to B-OHB and culturing for an additional 24 hours in control serum. Metabolism studies demonstrated that the early somite embryo possesses a limited capacity to oxidatively metabolize B-OHB. The biochemical implications of these findings are discussed with respect to the possible role of ketone bodies in the mechanism of diabetes-induced congenital malformations.
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  • 68
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    The @Anatomical Record 213 (1985), S. 448-456 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Proliferation of epithelial cells in the fetal trachea was studied in hamsters, beginning on the 10th gestational day and ending on the 16th day, shortly after birth. The mean mitotic index (MI) was highest on day 10, with no statistical confirmation of a change between days 10 and 11. The MI fell to about 2% on days 12 and 13, and declined thereafter to about 0.3% on day 16. The MIs for dorsal and ventral surfaces were compared and values were similar except on day 10, when ventral exceeded dorsal, and on day 12, when dorsal exceeded ventral, 2.56% and 1.3%, respectively. On days 10, 11, and 12 the epithelium was simple, composed of poorly differentiated columnar cells that proliferated. On day 13 the epithelium was pseudostratified owing to the presence of a few short cells that did not reach to the lumen. Throughout the fetal period, proliferation of columnar cells predominated but division of short (basal) cells increased from 8% to 40% of the total mitotic activity between day 13 and day 15. Proliferation of basal cells then declined, so that on day 16, 84% of all cells in mitosis were columnar. If basal cells divide to make more of themselves they must proliferate rapidly between day 13 and day 15, because they were virtually absent on day 12 but accounted for about 36% of the ventral and 23% of the dorsal epithelial cells on day 16. Based on the results, a hypothetical model is proposed for the formation of pseudostratified mucociliary epithelium. The model proposes that small granule (endocrine) cells, basal cells, secretory cells, and ciliated cells are established from poorly differentiated columnar cells by day 13. Thereafter short basal cells replicate to produce only basal cells, whereas columnar secretory cells replicate to produce secretory cells and ciliated cells. This model will be tested in future studies.
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  • 69
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    The @Anatomical Record 213 (1985), S. 464-472 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: White Leghorn chicken eggs, specific pathogen free, were treated with the organophosphate insecticide dicrotophos and the early defects thus induced were characterized histologically. Eggs were incubated for 24, 48, 72, or 96 hr, injected with doses of dicrotophos ranging from 250 μg to 2.0 mg per egg, and observed after an additional 48 hr of incubation. Treated embryos displayed general developmental retardation as well as unilateral retardation of the cranial sense organs, the youngest embryos being most severely affected. Many embryos injected with insecticide at 24 hr, and all but one injected at 48 or 72 hr, displayed notochordal folding, usually restricted to the cervical region; most of these also showed deformities of the adjacent spinal cord. Other defects, seen on a less consistent basis, included branching of the neural canal in the lumbar region, bifurcation of the neural epiphysis, deformation of the lens vesicle, and distention of the major blood vessels. The incidence and severity of epiphyseal, lens, and vascular defects were greatest among embryos treated at 24 hr, whereas notochordal and both types of neural defects were greatest among those treated at 48 hr. The incidence and severity of the abnormalities diminished with increasing age such that by 96 hr the only defect noted was a weak notochordal folding in one embryo. To a lesser extent, incidence and severity were dose-related also. Histological similarities between embryos displaying vascular distention and recently dead treated embryos suggested that this abnormality is a precursor to death. All defects were associated with the presence of the insecticide at the time the affected structures were undergoing initial or early morphogenesis or else the deposition of a supportive sheath, suggesting these activities as targets of the teratogen.
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  • 70
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    The @Anatomical Record 213 (1985) 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
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  • 71
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    The @Anatomical Record 213 (1985), S. 473-476 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Linear regression analysis was used to determine the correlation between testicular volume (calculated from measurements of length and width) and weights of paired testes. Line equations that can be used to predict weights of paired testes from the volume of one or both testes are given for three species of hamster (Mesocricetus auratus, Mesocricetus brandti, and Phodopus sungorus sungorus). Because these parameters are highly correlated (r ≥ 0.97), the line equations provide a highly reliable means of estimating weights of paired testes from in situ surgical measurements. Weights of testes predicted in this manner are highly correlated with actual weights of paired testes (r ≥ 0.96). The surgical method for measurement of the gonads is also described. This technique for estimating paired testicular weight allows considerable reduction in the number of animals required for longterm experiments and also permits longitudinal studies of individual animals.
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  • 72
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    The @Anatomical Record 213 (1985), S. 477-480 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The degree of metachromasia of mast cell granules is known to vary with the type of tissue fixation and among different tissues and species. The present study sought to determine whether mast cells in dog skin are heterogeneous with respect to fixation and staining properties. We performed skin biopsies in six anesthetized, atopic dogs and one mongrel dog. One biopsy was fixed in formalin and a second, from a parallel abdominal site, was fixed in basic lead acetate (Mota's solution). Adjacent sections from each biopsy were stained with alcian blue (1%, pH 0.5) or for chloroacetate esterase activity. In alcian blue-stained sections, one-third fewer mast cells were detected in skin fixed in formalin (1,836 ± 454 mast cells/mm3, mean ± SEM) than in skin fixed in basic lead acetate (2,684 ± 527 mast cells/mm3) (P 〈 0.05). The chloroacetate esterase reaction detected the larger number of mast cells regardless of the fixative used. We conclude that mast cell heterogeneity, as demonstrated by metachromatic staining following different types of tissue fixation, exists in dog skin. “Typical” mast cells stain with alcian blue regardless of fixation; however, “atypical” mast cells exhibit metachromasia only after fixation in basic lead acetate. Both the typical and atypical types of mast cells have chloroacetate esterase activity.
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  • 73
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    The @Anatomical Record 213 (1985), S. 481-495 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: In previous studies of the adult bat, Myotis lucifugus, bone loss that occurred during the hibernating period was attributed to osteocytic osteolysis, rather than osteoclastic activity. We have used histochemistry and light and electron microscopy to determine the functional state of the skeleton during the hibernating period. We find that during hibernation the marrow cavity of the long bones is filled with lipid deposits interspersed with vascular sinusoids containing mononuclear cells and red blood cells. The lipid deposits are found within fat cells and at extracellular sites. Hematopoietic cells are absent, osteoclasts are absent, and the bone surfaces are covered with quiescent bone-lining cells. Osteocytes retain their structural integrity and maintain canalicular systems with the bone lining cells and with other osteocytes.During the arousal period, osteoclasts reappear on the bone surface, followed in time by increased numbers of osteoblast-like cells. Perivascular cells undergo structural hypertrophy. Many mononuclear cells are now found in extravascular sites. The lipid content in the marrow is gradually reduced and replaced by hematopoietic cells. Each of these events occurs in a repeatable sequence that is related to the state of hibernation of Myotis lucifugus.
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  • 74
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    The @Anatomical Record 212 (1985) 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
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  • 75
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    The @Anatomical Record 212 (1985), S. 103-109 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Substance P immunoreactivity in the major pelvic ganglion (MPG) of the rat was studied to define a possible role for this neuropeptide in functions of the pelvic portion of the autonomic nervous system. Substance P immunoreactivity was found in three locations in the ganglion: (1) as a plexus of varicose fibers, (2) in small intensely fluorescent (SIF) cells, and (3) after colchicine pretreatment, in some principal neurons. The perineuronal plexus of fibers appeared as small varicosities closely related to the somae of principal neurons. Approximately 10-20% of principal neurons were enclosed by a substance P-positive plexus. SIF cells were intensely stained for substance P. The general relationships of SIF cells in this ganglion were confirmed by their staining for substance P: their occurrence singly or in large clusters, their short tapering processes often related to principal neurons, and the occasional presence of a beaded process. Colchicine treatment resulted in the appearance of rare principal neurons that stained for substance P. The pelvic nerve was surgically interrupted to determine whether the perineuronal plexus of varicose fibers had an intrinsic origin or arose from cell bodies outside the ganglion. The perineuronal plexus was virtually absent following this procedure. The results of this study indicate that principal neurons in the major pelvic ganglion may be subject to the influence of substance P derived from two sources: (1) intrinsic substance P-containing SIF cells and (2) neurons probably residing in dorsal root ganglia. The nature of principal neurons that acquire staining for substance P after colchicine is unclear.
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  • 76
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    The @Anatomical Record 212 (1985), S. 143-152 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The ultrastructure of globule leukocytes is described in the nasal and tracheal respiratory epithelium of three boys suffering from chronic airway infections. The globule leukocytes lie free in the intercellular spaces and appear to be migratory cells. They are characterized by intracytoplasmic membrane-bound globules, variable in number, size, shape, and internal structure. The globules may apparently release their content between the neighboring epithelial cells. Human globule leukocytes are also characterized by the presence of intracytoplasmic rod-shaped bodies, the significance of which is not known. They usually display an extended juxta-nuclear Golgi apparatus, presumably involved in the formation of the globules. Comparison of the fine structure of the globules in the globule leukocytes with that of the granules found in the subepithelial mast cells does not support a mast cell origin for human globule leukocytes. On morphological grounds, natural killer cells are postulated as a possible source for globule leukocytes. The function of globule leukocytes is briefly discussed. We presume that the globule leukocytes belong to the group of migrating and secreting cells involved in the defense of the organism against foreign material.
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  • 77
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    The @Anatomical Record 212 (1985), S. 132-142 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Amine precursor uptake and decarboxylation (APUD) small-granule cells were stained by periodic acid-Schiff (PAS)-lead hematoxylin in 0.5-μm etched Epon sections of adult hamster lung fixed for transmission electron microscopy. The leading edge of a small-granule cell cluster was identified in a segmental bronchus as a single PAS-positive cell. From 256 serial thin sections through its entirety, a three-dimensional wooden reconstruction of the cluster and morphometric estimates of the apical and basal surfaces, cell volume, and intracytoplasmic distribution of mitochondria and small granules was made. Of moderate size, the body consisted of 16 small-granule cells, 11 forming its ovoid core with five outlying cells diverging at the margin; these were pyramidal, possessing wide bases and thin apical processes. At the bronchial surface, processes from the 11-cell core emerged together, whereas the divergent cells emerged in groups of two and three. Ten Clara-like cells and one ciliated cell encircled the core Altogether they formed a pseudostratified epithelium in contrast to the surrounding simple columnar epithelium. Deeper in the cluster, numerous cytoplasmic extensions interdigitated with those from adjacent cells, and toward the base the Clara-like and APUD cells were increasingly interposed. In marked contrast to the apical cytoplasm, the infranuclear cytoplasm of the latter was densely packed with ca. 1,000 Å electron-dense granules; and the basal, presumptively secretory face of each cell was five to six times greater than the area exposed to the bronchial lumen. Judged by granule size and ultrastructure, only one APUD cell type was recognized in the reconstructed cluster. Beneath it many fibrocytic processes were separated from the APUD cells by only the thickness of the basal lamina. Two fascicles of smooth muscle approached the cluster within 0.4-0.8 μm. Unmyelinated nerve fibers came as close but contacted only the muscle. Capillaries, in contrast, came no closer than 15 μm from the base of the body. Evidently, (1) fibrocytes and smooth muscle are more likely targets for secretions from such a paracrine body than cells reached through the bloodstream, and (2) not all small-granule cell clusters are innervated.
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  • 78
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    The @Anatomical Record 212 (1985), S. 161-166 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Discovery of components of the renin-angiotensin system (RAS) in the adenohypophysis of several species has prompted speculation concerning the location and possible function of a pituitary RAS. Although both renin and angiotensin II have been localized within the rat adenohypophysis, their colocalization has not been previously demonstrated within the same cells. In the present study, immunohistochemical staining by the avidin-biotin-peroxidase complex technique was used to demonstrate the coexistence of renin and angiotensin II in adenohypophyseal cells identified morphologically and immunocytochemically as gonadotrophs. These results support the existence of an adenohypophyseal RAS, at least part of which is under intracellular control. The influence of this system on control of fluid balance, blood pressure, and the secretion of other hypophyseal hormones is discussed.
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  • 79
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    The @Anatomical Record 212 (1985), S. 167-171 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A method for the estimation of the size and total number of calcitonin-containing cells (C cells) in the rat thyroid gland has been devised. The total area, the number of C cells per unit area, and the areal fraction of C cells were determined for the C cell region using step serial sections. From these data it was estimated that from 0.3 × 106 to 1.0 × 106 C cells were evenly divided between the two thyroid lobes. Approximately 150 μm3 of cytoplasm were associated with each of these cells. In comparison with sham-operated rats, pinealectomy had little effect on the number of C cells. In an experiment terminated in the summer, there was a statistically insignificant decrease 6 weeks postsurgery; no effect was seen at 12 weeks. On the other hand, a slight increase in the number of C cells was seen in January, 12 weeks postsurgery. The volume of cytoplasm per cell was not altered by pinealectomy.
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  • 80
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    The @Anatomical Record 212 (1985), S. 179-182 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Pial surface arterioles join rami of the anterior and middle cerebral arteries (MCA) in 10-day-old normal Wistar rats. Normally, the anastomosing branches differentiate into small-diameter vessels with significantly greater tortuosity (length) by 56 days of age. After interruption of the MCA in 10-day-old rats, large-diameter, relatively straight collaterals were observed 46 or 90-110 days later. Evidently altered hemodynamic factors, as the result of MCA interruption, prevent the development of a normal pattern of arterioles and augment the development of an anomalous pattern of collaterals in young Wistar rats.
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  • 81
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    The @Anatomical Record 212 (1985), S. 195-198 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Patten described as neural overgrowth a folding of brain tissue into the ventricles in abnormal human embryos. Intubation of chick embryo brains releases cerebrospinal fluid pressure resulting in a similar folding. These folded brains have 2.5 times less tissue volume than control embryos of identical stage. This study shows, with total DNA measurements, that brain cell number is 50% less in folded brains (intubated embryos) as compared to control brain of identical staged embryos.
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  • 82
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    The @Anatomical Record 212 (1985), S. 183-194 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Previous investigation into the regenerative ability of postmetamorphic adult land phase Ambystoma has revealed that (1) these species have the capacity to completely regenerate a limb, given optimal environmental conditions, and (2) the gross morphological characteristics of limb regeneration in these species compared favorably with the external regeneration morphology of aquatic phase forms. The present study concerns a histological and histochemical examination of the regenerating limb tissues and their respective extracellular and intracellular tissue matrices.Postmetamorphic adult Ambystoma were amputated through the forearm, placed within optimal environmental conditions, and allowed to regenerate. The tissues were harvested at designated intervals after amputation and prepared for light microscopic examination. The limb tissues were assayed histologically for similarities to and differences from previously established regeneration morphologies. It was noted that specific correlations (i.e., apical epidermal cap formation, bud outgrowth and elongation, palette formation, and digit formation) existed between regeneration histologies in these species and those previously reported for the aquatic urodeles, newt, axolotl, and larval salamander. By utilizing the histological and histochemical characteristics of the tissue, the regenerate limb was divided into five tissue units: epidermal, blastemal, soft, hard, and neuro/vascular. Based on the unique morphology of their extracellular matrices and respective histochemical staining patterns, four distinct blastemal regions were delineated within the blastemal units: subregenerate epidermal blastema, soft-tissue blastema, hard-tissue blastema, and core blastema. Histochemically, changing patterns of highly sulfated, weakly sulfated, and carboxylated polysaccharides and glycosylated compounds were located within both the extra- and intracelluler stump and regenerate tissue matrices during regeneration. In addition, these patterns of intra- and extracellular macromolecular material correlated to previous reports of similar-type compounds assayed during regeneration in aquatic urodeles. With this in mind, the adult land phase Ambystoma can be considered an appropriate model system for studies concerning normal limb regeneration.
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  • 83
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    The @Anatomical Record 212 (1985), S. 199-205 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Cell differentiation in the retina of the mouse during the postnatal period was studied by autoradiography. Animals were injected with 3H-thymidine at ages extending from the day of birth through postnatal day 11. Six weeks later the distribution of labeled nuclei in the cells of the mature neural retina was analyzed to determine when these cells completed their final mitosis prior to differentiating. Central and peripheral zones were analyzed separately. Cell division ceases by 5-6 days in the center of the retina and by 11 days in the periphery. Among cells produced postnatally, 73% differentiate as rods, 20% as bipolar cells, 6% as Müller cells, and 1% as amacrine and ganglion cells. At all stages of embryogenesis, the differentiation of at least three and as many as six distinct types of specialized cells is initiated simultaneously within the ventricular cell population.
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  • 84
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    The @Anatomical Record 212 (1985), S. A225 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
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  • 85
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: In order to elucidate the functional significance of somatostatin in thyroid C cells, the alterations of immunoreactive somatostatin in the cells were investigated under various experimental conditions, i.e., hypercalcemia, hypocalcemia, and antithyroid drug treatment. Guinea pigs and rabbits, in which almost all C cells reveal the intense immunoreaction for somatostatin in addition to calcitonin, were used as experimental animals. After chronically induced hypercalcemia, somatostatin immunoreactivity conspicuously diminished coinciding with the decrease of calcitonin; somatostatin as well as calcitonin was responsive to induced hypercalcemia. After hypocalcemic tetany induced by injection of Escherichia coli Lasparaginase, C cells exhibited very intense immunoreactions for both calcitonin and somatostatin. After chronic treatment of ethylenethiourea, immunoreaction of somatostatin in C cells was the same as that of calcitonin. That is, when immunoreactivity for calcitonin remained unchanged, immunoreactivity for somatostatin was also intensive. However, when immunoreaction of calcitonin became very weak, the reaction of somatostatin was also weak. Thus, in all experimental conditions examined the alterations of immunoreactive somatostatin in C cells completely coincided with those of calcitonin. It seems likely that somatostatin in thyroid C cells exerts the synergistic effect on calcitonin action.
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  • 86
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    The @Anatomical Record 211 (1985), S. 285-294 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Ciliated cells of the ductuli efferentes show at their apex a discrete endocytic apparatus composed of small vesicles connected to or subjacent to the apical plasma membrane, small apical membranous tubules, and pale multivesicular bodies. Deeper in the cytoplasm, there are acid phosphatase-positive denser, multivesicular bodies and secondary lysosomes showing an electron-dense cortex and a crystalline, paler stained core. Cationic ferritin and concanavalin A-ferritin used to demonstrate adsorptive endocytosis, when injected into the rete testis, rapidly reached the lumen of the ductuli efferentes. At 1 min after injection, these tracers were seen bound to the apical plasma membrane of ciliated cells and within small endocytic vesicles and by 5 min in narrow apical tubules. At 15 and 30 min after injection, the tracers appeared in pale multivesicular bodies while at 1 hr they were found within dense multivesicular bodies. At 2 hr and longer time intervals these tracers accumulated within secondary lysosomes. Native ferritin, concanavalin A-ferritin in the presence of α-methyl-D-mannoside, and horseradish peroxidase or albumin-colloidal gold complexes were used to analyze fluid-phase endocytosis. At various intervals following their injection into the rete testis, these tracers presented a distribution identical in all respects to that described for cationic ferritin and concanavalin A-ferritin. In the present work, none of the above tracers were transported to the abluminal aspect of the ciliated cells. These cells, like the nonciliated epithelial cells of the ductuli efferentes are thus involved in adsorptive as well as in fluid-phase endocytosis.
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  • 87
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    The @Anatomical Record 211 (1985), S. 295-303 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: This study systematically characterizes the organization and nature of epithelial populations in the distal airways of the adult rhesus monkey. Infusionfixed lungs were evaluated using airway dissection and scanning and transmission electron microscopy. We found that a true bronchiole free of cartilage and alveolar outpockets was not consistently present. Cartilage and alveolar outpocketings were often observed within the same airway generation. The epithelial population of nonalveolarized terminal conducting airways was pseudostratified columnar, consisting of ciliated, mucous, and basal cells. The respiratory bronchiole found immediately distal to the terminal conducting airways had two clearly demarcated zones of distinctly different epithelial populations. Overlying the pulmonary artery was the same pseudostratified ciliated columnar epithelial population observed in nonalveolarized terminal airways. The epithelial population in the remainder of the respiratory bronchiole, not associated with the pulmonary artery, was simple nonciliated cuboidal with a few squamous cells. The cuboidal nonciliated bronchiolar cell differs from the mucous cell by having few small granules and rough and smooth endoplasmic reticulum. The extension of the ciliated, mucous, and basal cells several generations into the respiratory bronchiole in rhesus monkey has not been observed in rodents and other laboratory mammals. Data from studies of human airways, although not explicit, suggest that the rhesus monkey epithelial cell pattern resembles the pattern in the human terminal airways.
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  • 88
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    The @Anatomical Record 211 (1985), S. 357-366 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: No Abstracts.
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  • 89
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    The @Anatomical Record 211 (1985), S. A1 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: No Abstracts.
    Type of Medium: Electronic Resource
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  • 90
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    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 211 (1985), S. A46 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: No Abstracts.
    Type of Medium: Electronic Resource
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  • 91
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    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 212 (1985), S. 255-267 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Muscle spindle ultrastructure of the extensor digitorum communis of pigeons was studied using spaced serial sections. The intrafusal fibers extended well beyond the ensheathing capsule, after which they became disoriented and often fused with each other before terminating on the connective tissue of extrafusal fibers. Several extracapsular fibers contained macrofibrils which were about 0.1 μm in diameter and contained several dozen smaller (10 nm) subunits. Intrafusal fibers commonly formed close attachments with one another for short or extended (240 μm) lengths. A basal lamina was absent between regions of pairing, and a myosatellite cell lay at the border of the coupled region. Several fibers could be coupled together in a single cross section; fibers coupled together, separated, and either recoupled or became associated with other fibers along the length of a spindle.Profiles of sensory terminals and sensory satellite cells alternated to form a smooth-contoured surface over most of the fiber cross section in the equatorial region. The sensory terminals contained many mitochondria, lysosomes, and clear and dense core microvesicles. Both the terminals and sensory satellite cells formed desmosomelike junctions with the intrafusal fiber. A crescentic collagen sheath covered that portion of the fiber cross section containing the sensory terminal-satellite cell complex. Inner capsule cells surrounded the entire assembly in the equatorial region.The basal lamina thicknesses differed over the naked intrafusal fiber compared to that portion covered by the sensory terminal or sensory satellite cell. The thickness was more than doubled over the latter regions, indicating that the basal lamina over these areas was a product of the fused intrafusal fiber and sensory terminal and/or sensory satellite cell basal laminae. These are discussed in terms of intrafusal fiber degeneration and regeneration.
    Additional Material: 14 Ill.
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  • 92
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    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 212 (1985), S. 277-281 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Guanylate cyclase activity has been studied biochemically and cytochemically in guinea pig testis. The results of the biochemical assays indicate an equal distribution of this enzyme between the soluble and particulate fractions, which have a different sensitivity to adenosine triphosphate. The cytochemical results demonstrate that the reaction product of guanylate cyclase is detectable in the interstitial capillary endothelial cells and, in the seminiferous epithelium, mainly at the level of the adjacent surfaces of Sertoli and germ cells of the intermediate and adluminal compartments. Guanylate cyclase activity appears at the level of pachytene spermatocytes and persists throughout subsequent stages of development. The distribution in the seminiferous epithelium seems to indicate that guanylate cyclase is involved in the interrelationships between Sertoli and germ cells during gamete differentiation.
    Additional Material: 10 Ill.
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  • 93
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The distribution of glucagon and pancreatic polypeptide was studied immunocytochemically in rat pancreas at both light and electron microscopic levels. My earlier observation that these two peptides are distributed in three cell types - cells containing glucagon, cells containing pancreatic polypeptide, and cells containing both - was confirmed at the electron microscopic level. In the glucagon-pancreatic polypeptide cells, the immunoreactivities of the two peptides were present in the same secretion granules. In addition, these glucagon and pancreatic polypeptide-containing granules were morphologically distinct from glucagon granules but similar to pancreatic polypeptide granules and somatostatin granules.
    Additional Material: 6 Ill.
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  • 94
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A quantitative autoradiographic immunocytochemical study was performed in which the nuclear uptake and retention of 3H-estradiol (3H-E2) by luteinizing hormone (LH) and prolactin (PRL) cells was examined in 19-21-year-old baboons. 3H-E2 concentrating cells were found in all of the three lobes of the pituitary in varying percentages (38.7%, pars distalis; 17.1%, pars intermedia; 6.3%, pars nervosa). Approximately 80% of PRL cells and nearly 100% of LH cells were labeled. A count of the number of silver grains over nuclei revealed a marked variation of the accumulation of 3H-E2 by LH cells and to a lesser extent in PRL cells. These results suggest functional heterogeneity among LH and PRL cells. The present results are discussed in relation to the physiological state of old animals.
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  • 95
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    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 211 (1985), S. 149-155 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A scanning electron microscope cast technique was used to examine the interface between primary and secondary dentine in young and old human teeth. In addition, more traditional methods were used to examine this interface with light microscopy, identical regions being viewed before and after demineralization. No continuity was seen between the tubules in primary dentine and those in irregular secondary dentine in the scanning electron microscope preparations. These preparations did show the tubules to be continuous between primary dentine and regular secondary dentine in young and old teeth. Both the scanning electron microscopic and light microscopic observations suggested that regular secondary dentine becomes highly sclerosed in old teeth.
    Additional Material: 10 Ill.
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  • 96
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    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 211 (1985), S. 142-148 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Samples of human plantar and palmar skin were excised and incubated in 20 mM EDTA after which the epidermis was gently separated from the dermis with the plane of separation occurring in the lamina lucida. Scanning electron microscopic examination of the dermal component revealed the classically described series of regularly spaced grooves and papillae that characterize the epidermal-dermal junction in thick skin. Primary dermal grooves exhibited evenly spaced tunnels that were originally occupied by sweat gland ducts. The basement membrane (basal lamina) in the primary grooves was relatively smooth but did exhibit a flattened, reticulated pattern at high magnifications. The basement membrane of secondary dermal grooves and papillae was in the form of numerous, elevated microridges off of which septae arose at roughly right angles. The surface appearance of the basement membrane in these areas was that of a honeycomb owing to the numerous compartments and recesses formed by the ridges and septae. Degradation of the basement membrane by trypsin demonstrated that the foundation for the highly folded and compartmentalized basement membrane was composed of dermal collagen fibrils, 60-70 nm in diameter, that were arranged in a series of variably sized, interconnected collagen bundles or walls. Epidermal basal cells extended cytoplasmic (foot) processes into two or more compartments, formed by the ridges and septae, which considerably amplified the basement membrane surface available for epidermal attachment. Scanning electron microscopic studies of the epidermal-dermal junction confirm the variable surface character of this interface previously reported by others using sectioned material. This regional variation in surface architecture apparently distinguishes between areas in which epidermal basal cells are specialized for attachment (papillae, secondary dermal grooves) and regions occupied by slow cycling epidermal stem cells from which mitotically active keratinocytes arise.
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  • 97
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    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 211 (1985), S. 156-165 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Postamputational healing was compared in nonregenerating and regenerating animals to determine whether bone healing might interfere with a regenerative response in mice. More than 150 mouse toes and 100 newt limbs were examined at the light microscope level. Stages of normal bone healing with approximate times of occurrence were established. Major differences in healing of these two species were seen. The periosteum produced hyaline cartilage, woven bone, and chondroid bone in mice, but only hyaline cartilage in newts. The endosteum produced woven bone in mice but no new growth in newts. Dead bone persisted in mice but was removed in newts. The marrow cavity became sealed in mice but remained open in newts. Despite these differences both animals produced skeletal tissue distal to the amputation plane. Woven bone formed distal to the amputation plane of mice. Cartilage formed distal to the amputation plane of newts, but cartilage was never seen distal to the plane of mice. Results of previous studies reveal that cartilage can be formed distal to the amputation plane of experimentally treated mice. Thus, although it does not regenerate, mouse bone is capable of producing, distal to the amputation plane, the type of skeletal tissue which appears at that location during an epimorphic regenerative response. This observation, in combination with other experimental results, indicates that both skeletal and soft tissues at the amputation site of treated mammals can resemble comparable tissues of newt limbs at an early stage of regeneration.
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  • 98
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    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 211 (1985), S. 166-173 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: During the middle of prenatal day 10, the opossum allantois forms as a ventral outgrowth of the hindgut. By day 11 it appears as a large, fluid filled sac and by the middle of day 12 (just prior to birth) it reaches its maximal development. The simple squamous epithelium lining the allantois consists of only one cell type that often contains numerous filaments in the apical cytoplasm. At the luminal surface, the apices of the cells are united by junctional complexes and desmosomes are present between adjacent cells. The luminal surface is irregular, whereas laterally and basally the cell membranes show few if any infoldings. Mitotic figures and presumptive degenerating cells occasionally occur in the allantoic epithelium which rests on a delicate basal lamina. The allantois is covered by a simple squamous mesothelium that lacks a distinct basal lamina. Between the two epithelial sheets lie mesenchymal cells, collagen fibers, and blood vessels. No specializations of cell membranes were noted in either of the epithelial layers. The yolk sac endoderm consists of a single layer of squamous cells whose cytoplasm contains scattered profiles of rough endoplasmic reticulum and mitochondria. Extensive lateral and basal infoldings of the plasmalemma were not observed in these endodermal cells. The morphology of the trophectodermal (trophoblastic) cells indicates a cell type that is active in the transport of materials. Since the endodermal cells that line the allantois lack morphological features that would suggest the presence of mechanisms for transport or exchange, and because they remain relatively unchanged throughout pregnancy, it is thought that the allantois functions primarily in the storage of urinary wastes during prenatal life. As the allantois does not establish physical contact with the yolk sac placenta, it is believed that exchange between the developing fetus and mother occurs primarily through the activity of cells that form the trophoblast.
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  • 99
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    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 211 (1985), S. 427-433 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Alveolar regions of normal pig lungs (newborn to 60-day-old) were characterized morphometrically to provide a basis for comparison in future investigations of porcine respiratory diseases. Endotracheal installation of fixative was done to expand the lungs uniformly at total capacity. Differential effects of lobar variations were determined by stratified random sampling of lung lobes. A stereologic study was done by point and intersection counts on electron micrographs. At birth, the lungs were remarkably well developed. Relative alveolar and capillary surface densities and air-blood tissue barrier thicknesses were at adult levels. In allometric regressions, volumes and surfaces of lung components regressed directly to lung volume, but monoexponentially (to the 3/4 power) with body weight. In the first postnatal week, however, relative volume densities of cellular interstitium in septal tissue and of capillary lumina in parenchyma increased at statistically significant levels. Composition of lung parenchyma and septa was changed, although without statistically significant direct impact on parameters related to gas exchange. Type II pneumocytes had increased nuclear to cytoplasmic volume ratios in 7- to 14-day-old pigs, probably reflecting cell activation and increased surfactant production. Age (postnatal lung growth) created the most substantial variance of results; interanimal variation in pigs of the same age was less important and no consistent lobar variations were seen.
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  • 100
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    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 212 (1985), S. 327-335 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Glycoprotein components of epidermal mucous cells in control (pH 6.8) and acid-stressed (pH 5.8, 4.8, 4.0) brown bullhead catfish were studied by histochemical and autoradiographic methods. Following exposure to acid, increased numbers of epidermal mucous cells were noted in all tissue sites studied, particularry dorsal and ventral skin. Mucous cells of control fish and acid-stressed (pH 5.8) fish contained a mixture of neutral and acidic mucosubsf ances, the latter including both sialomucins and sulfomucins, with sialomucins predominating. An apparent shift toward increased sulfomucin production was observed in skin from fish exposed to pH 4.8 and 4.0. In addition, autoradiographic studies using 35S showed increased labeling of mucous cells in acid-exposed (pH 4.0) epidermis, offering further support for a shift toward sulfomucin production in acid-stressed fish. Absence of appreciable sulfomucin production in younger fish (〈1 year old) suggested that this shift may be partly age-dependent. The functional significance of increased sulfomucin production in acid-stressed epidermal mucous cells is uncertain but may be related to maintenance of ionic equilibrium.
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