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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 168 (1976), S. 101-115 
    ISSN: 1432-0878
    Keywords: Synapses ; Microtubules ; Albumin ; Tissue culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary This paper describes new ultrastructural features of neural processes and of synapses in cultured CNS tissue treated with albumin before fixation using a modification of the technique recently introduced by Gray (1975). Nerve fibre bundles in explants of foetal spinal cord grown in vitro for 15–18 days were transected microsurgically. After transection the cultures were exposed to 20% albumin in distilled water and then fixed in unbuffered osmium tetroxide followed by unbuffered glutaraldehyde. In this material, but not in controls (injured but not exposed to albumin; exposed to albumin without injury) microtubules were found within many axonal varicosities, often situated close to presynaptic membrane specializations. These microtubules were closely associated with vesicles resembling synaptic vesicles, which were occasionally aligned in rows along the microtubules. Similar vesicle-microtubule associations were also found in non-terminal axons. Microtubules were also observed very close to some postsynaptic densities. The possibility that the microtubule-vesicle associations are involved in vesicle movements (along axons and/or within axon terminals) is discussed. A more direct involvement of microtubules in terminals in the mechanism of transmitter release is also considered.
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  • 2
    ISSN: 1432-0878
    Keywords: Neuromuscular junctions ; Development in vitro ; Electron microscopy ; Electrophysiology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The development of neuro-muscular junctions between previously dissociated foetal rat spinal cord and somatic muscle has been investigated. The first indications of junction formation, both ultrastructurally and electrophysiologically, were observed after circa 18 days in vitro. The junctions contained numerous vesicles, but no secondary folds were developed even after 6 weeks in culture, and synaptic densities were not well marked. Functional endplates were found, and action potentials, endplate potentials and miniature endplate potentials recorded.
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 169 (1976), S. 41-47 
    ISSN: 1432-0878
    Keywords: Dorsal root ganglia ; Tissue culture ; Microtubules ; Axons
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Microtubule fascicles, resembling those characterizing the initial segment of multipolar neurons, have been observed by electron microscopy within and close to the origin of the stem process of some unipolar ganglion cells in explant cultures of embryonic chick dorsal root ganglia. Each fascicle comprised 2–6 closely spaced parallel microtubules linked by electron dense cross-bridges. Since similar observations have been made on stem processes in vivo, the possibility that linked microtubules occur commonly in this site is considered. The observations are discussed in relation to a possible correlation between the presence of microtubule fascicles and the initiation of action potentials.
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 140 (1973), S. 203-216 
    ISSN: 1432-0878
    Keywords: Synapses ; Development ; Chick embryo ; Cell culture ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The formation and development of synaptic contacts between dissociated chick spinal cord neurons has been investigated. By the 6th day in vitro “immature” profiles with few vesicles were observed. By 14–18 days “mature” types with numerous vesicles were found, indistinguishable from those of newly hatched chick spinal cord. After this period degeneration occurred, and was especially marked in the post-synaptic element. Such degeneration could be postponed by the addition of small numbers of somatic muscle cells. The Kanaseki and Kadota (1969) technique was applied to the study of coated vesicles at various stages of synaptic development.
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  • 5
    ISSN: 1432-0878
    Keywords: Tannic acid ; Acetylcholine receptors ; Tissue culture ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Spinal cord neurons from 9-day chick embryos were maintained in culture for up to 35 days and then fixed in 4% cacodylate-buffered glutaraldehyde containing 2% tannic acid. After about 15 days in culture a small percentage of the synaptic specializations present were characterized by striking electron-dense striations averaging 15 nm in width, oriented perpendicular to the postsynaptic membrane. These structures increased in frequency with time in culture (to a maximum of about 10% of all synapses in the oldest cultures); they were asymmetrical, protruding approximately 8 nm into the synaptic cleft, and more deeply (approximately 15–18 nm), into the postsynaptic cytoplasm. On the basis of earlier work by Sealock (1980) they are interpreted as concentrations of acetylcholine receptors. Similar membrane differentiations were also seen associated with active-zone areas of a few presynaptic membranes, and the possibility that these represent presynaptic acetylcholine receptors is discussed. Additional observations reported are (1) the presence of striations resembling those seen at the postsynaptic membrane in the membranes of some postsynaptic vesicles, and (2) filamentous links between the striations and cytoskeletal elements of the postsynaptic cell.
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 245 (1986), S. 563-577 
    ISSN: 1432-0878
    Keywords: Chick retina ; Tissue culture ; Electron microscopy ; Development, ontogenetic ; Differentiation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The differentiation of cells and synapses in explants of 9-day-old chick embryo retina has been studied by light and electron microscopy over a period of 35 days in vitro, and samples of retina from the 9-day chick foetus were directly fixed and prepared for study. At the time of explantation the retinae were poorly differentiated and no lamination was apparent. From day 14 onwards, (i) outer and inner nuclear layers (ONL, INL) separated by a layer of neuropil corresponding to the outer plexiform layer (OPL) and (ii) a layer of scattered large ganglion cells separated from the INL by a zone of neuropil resembling the inner plexiform layer (IPL) were apparent, and (iii) a well-differentiated outer limiting membrane was established close to the surface of the explants. In the oldest cultures some development of photoreceptor outer segments occurred but a distinct optic nerve fibre layer did not form. Although cell identification presented problems even in the oldest cultures, the major retinal cell types described in vivo could be identified. Photoreceptor cells developed pedicles in the OPL which became filled with synaptic vesicles and synaptic ribbons and established ribbon synapses (including triads) with and were commonly invaginated by processes from horizontal and bipolar cells. Processes of bipolar cells in the IPL formed simple and dyad synapses. At least two types of presynaptic amacrine cells were also identified in the INL, one of which contained large numbers of dense-core vesicles. The ganglion cells, though sparse, were large and well differentiated. These findings show that all the major neuronal types of the retina are capable of developing and differentiating in vitro, lagging behind the time-table of development and differentiation in vivo by approximately 7 days, but resulting in a histotypically organised retina with synaptic neuropil showing many similarities to the corresponding neuropil in vivo.
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  • 7
    ISSN: 1432-0878
    Keywords: Explant cultures ; Synaptic morphology ; Magnesium-rich medium ; Mouse
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The present study examines the ultrastructure of synaptic profiles developed in explant cultures of immature mouse cerebral cortex, maintained for prolonged periods in a magnesium-rich environment. The ethanolic phosphotungstic acid method was employed in addition to conventional preparation procedures so that paramembranous densities could be clearly observed. Although presynaptic terminals were frequently packed to capacity with vesicles in cultures maintained in magnesium-rich media, there was always a proportion which contained loosely collected vesicles. Few other changes in synaptic morphology were apparent and the paramembranous densities were unaffected. The reasons for the effectiveness of the transmission block, the absence of any change in the morphology of paramembranous densities, and the excessive crowding of presynaptic vesicles are considered and discussed.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 190 (1978), S. 525-538 
    ISSN: 1432-0878
    Keywords: Nerve fibre bundles ; Chick embryo ; Transection ; Effects on axons, cones, glia cells ; Microsurgery in vitro
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Fibres growing from neurons of explanted dorsal root ganglia from 10 day chick embryos were transected and subsequently observed by light and electron microscopy after periods of a few to fifty minutes. Changes immediately proximal and distal to the cut together with alterations further away from the site of injury on both sides of the cut were recorded. Observations were also made on the growth cones of damaged axons and on changes in associated glial cells. Reactive and degenerative changes including the rotation, retraction and swelling of cut axons occurred rapidly. Electron microscopy revealed tracts of filamentous material close to the sealed-off ends of axons, swollen organelles such as mitochondria, and lamellar bodies of varying dimensions. Proximal to the injury and closer to the expiant, damaged and degenerating axons mingled with normal processes. Many contained only a fine granular material, others clumps of organelles, particularly mitochondria. Distal to the cut, microspikes were lost from some growth cones. The dense granular material filling microspikes and growth cones remained unchanged. Clumps of large clear vesicles, lamellar bodies and swollen degenerating mitochondria were present, not only within growth cones, but also in all parts of the axon distal to the cut. Glial cells associated with transected axons soon developed an electron dense cytoplasm containing swollen organelles. Large numbers of vesicles filled with a particulate substance were also found. The possible significance of the changes observed after transection are considered and discussed.
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 194 (1978), S. 503-511 
    ISSN: 1432-0878
    Keywords: Cell cultures ; Electron microscopy ; Synaptic organelles ; Spinal cord (mouse)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary This study describes some of the ultrastructural features of presynaptic and postsynaptic organelles at synapses developed in cultures of previously dissociated mouse spinal cord cells. Particular attention was paid to the agranular reticulum which is well developed at many presynaptic and postsynaptic sites, either in the form of simple tubules or cisternae, or more complex networks and often closely associated with mitochondria. In addition, the disposition of microtubules at and close to synaptic specializations is described. These and other features of synaptic zones, such as granular vesicles in presynaptic sites, are discussed in relation to cultures developed on feeder layers and synapses in vivo, and in relations to possible degenerative and regenerative events in the cell cultures.
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 183 (1977), S. 403-417 
    ISSN: 1432-0878
    Keywords: Cerebral cortex ; Human foetus ; Synaptogenesis ; Cell culture ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Cells from foetal human cerebral cortex were mechanically dissociated and subsequently maintained in vitro for periods ranging between three and twenty-eight days. The ultrastructure of these cells at different stages of their development in culture was extensively examined. Nuclear and cytoplasmic features were extremely variable and a wide range of cell types was evidently represented. Of the three principal cell types found i.e. neurons, neuroglia and mesenchymal cells, only a minority of cells was classified with confidence, particularly during the first two weeks in culture. Extensive intercellular junctions of the adhaerens variety, common after 14 days in vitro were present at an earlier stage of development than synaptic profiles. First indications of synapse formation were observed after 21 days in vitro and after 24 days presynaptic sites filled with synaptic vesicles and with well defined presynaptic and postsynaptic thickenings were found. The significance of some of the features observed are both considered and discussed.
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