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  • 1
    Electronic Resource
    Electronic Resource
    The development and ultrastructure of previously dissociated embryonic chick corpus striatum cultured on feeder layers of liver cells (1980)
    Springer
    Anatomy and embryology 159 (1980), S. 115-123 
    Details
    ISSN: 1432-0568
    Keywords: Corpus Striatum neurons ; Cell culture ; Liver feeder layers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Embryonic chick corpus striatum neurons were dissociated and maintained on liver feeder layers in culture. Although some large dark-cored vesicles were present in many nerve processes and presynaptic boutons they were substantially less numerous than chick spinal cord neurons grown under identical conditions. Paraformaldehyde-induced fluorescence, although observed in a few culture batches in aggregates of corpus striatum neurons, was otherwise absent and no decisive evidence was obtained to suggest that fluorescent corpus striatum neurons were commonly developed on liver feeder layers in culture. Microtubules filled most cell bodies and nerve processes, and extended well into synaptic boutons often approaching the active zones. They were much more abundant in cultures of corpus striatum than in comparable spinal cord preparations and formed the principal organelle of many nerve fibres. These differences between chick spinal cord and corpus striatum neurons are both interesting and difficult to interpret. It is possible that fewer appropriate cholinergic neurons are available for transformation into adrenergic neurons within the corpus striatum, and that excessive numbers of dark-cored vesicles indicate only a greatly increased rate of acetylcholine production and storage.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00299260
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  • 2
    Electronic Resource
    Electronic Resource
    The development of fluorescent neurons after noradrenaline and dopamine are added to embryonic chick forebrain cells in vitro (1981)
    Springer
    Anatomy and embryology 163 (1981), S. 235-244 
    Details
    ISSN: 1432-0568
    Keywords: Chick embryos ; Forebrain neurons ; Cell culture ; Fluorescence ; Transmitters
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary In contrast to embryonic chick spinal cord neurons maintained on liver feeder layers in culture, chick forebrain cells from similar embryos treated in the same way failed to develop paraformaldehyde-induced fluorescence or large numbers of dense-cored vesicles. However, after the addition of noradrenaline, 5–8% of forebrain neurons did develop both the above features which are characteristic of adrenergic neurons. A less marked effect was seen with the addition of dopamine (circa 1% of the cells showed fluorescence). The differences between spinal cord and forebrain cells in culture are thought to reflect in part, considerable differences between them in development and maturity. The liver feeder layers probably play a vital role, inducing the changes within both CNS regions and thus a change in the phenotype of some neurons. In embryonic chick forebrain cells the addition of noradrenaline or dopamine may be necessary only because of the comparative immaturity of the cells. Factors influencing the distribution of fluorescence and the location of dense-cored vesicles are considered and discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00320679
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  • 3
    Electronic Resource
    Electronic Resource
    The development of synapses in vitro between previously dissociated chick spinal cord neurons (1973)
    Springer
    Cell & tissue research 140 (1973), S. 203-216 
    Details
    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.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00306695
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  • 4
    Electronic Resource
    Electronic Resource
    An ultrastructural and electrophysiological study of the development of neuro-muscular junctions between previously dissociated foetal rat cells in vitro (1974)
    Springer
    Cell & tissue research 155 (1974), S. 269-282 
    Details
    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.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00221359
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  • 5
    Electronic Resource
    Electronic Resource
    Microtubule fascicles in the stem processes of cultured sensory ganglion cells (1976)
    Springer
    Cell & tissue research 169 (1976), S. 41-47 
    Details
    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.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00219306
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  • 6
    Electronic Resource
    Electronic Resource
    Microtubule-synaptic vesicle associations in cultured rat spinal cord neurons (1976)
    Springer
    Cell & tissue research 168 (1976), S. 101-115 
    Details
    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.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00219727
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  • 7
    Electronic Resource
    Electronic Resource
    Regions of putative acetylcholine receptors at synaptic contacts between neurons maintained in culture and subsequently fixed in solutions containing tannic acid (1984)
    Springer
    Cell & tissue research 235 (1984), S. 85-89 
    Details
    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.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00213727
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  • 8
    Electronic Resource
    Electronic Resource
    The morphology of synaptic profiles in expiants of foetal and neonatal mouse cerebral cortex maintained in a magnesium-enriched environment (1980)
    Springer
    Cell & tissue research 206 (1980), S. 115-122 
    Details
    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
    URL: http://dx.doi.org/10.1007/BF00233612
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  • 9
    Electronic Resource
    Electronic Resource
    Ultrastructural observations on rapid formation of neuro-muscular junctions in vitro (1981)
    Springer
    Cell & tissue research 217 (1981), S. 647-659 
    Details
    ISSN: 1432-0878
    Keywords: Cell culture ; Neuro-muscular junctions ; Short-term preparations ; Mouse ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The development of neuro-muscular junctions (mouse, rat) from the time of first contact between neurons and myotubes in culture and the changes which lead to the formation of functional synaptic contacts have been investigated using light microscopy and ultrastructural techniques. An extensive basal lamina was present when the neuronal cell population was added to the developing myotubes in culture. The nerve cells were initially strongly attracted to each other and nerve cell aggregates formed rapidly. It was only when nerve fibres began to grow out of these aggregates to contact developing myotubes that changes within the cytoplasm of the two adjacent cells were observed. These developments included accumulations of filaments, membrane densities, mitochondria and large clear vesicles within both cells in the region of contact. In addition, collections of glycogen granules and an extensive membrane reticular complex were found within myotubes, and an extensive granular material filled many of the nerve processes. The basal lamina within the intercellular space appeared more electron-dense than elsewhere and was traversed by strands linking the two cell membranes. These features all appeared to be stages in the initial formation of neuro-muscular junctions. It was only after these events had occurred that presynaptic vesicles gradually appeared within the future nerve terminal. The results of this paper therefore support the view that synaptic transmission at developing mammalian neuromuscular junctions is not necessarily dependent on the presence of presynaptic vesicles.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00219371
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  • 10
    Electronic Resource
    Electronic Resource
    Cellular uptake of the prion protein fragment PrP106-126 in vitro (1999)
    Springer
    Journal of neurocytology 28 (1999), S. 149-159 
    Details
    ISSN: 1573-7381
    Keywords: prion disease ; transmissible spongiform encephalopathy ; cell culture ; toxicity ; apoptosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The aetiological agent of prion disease is proposed to be an aberrant isoform of the cell surface glycoprotein known as the prion protein (PrPc). This pathological isoform (PrPSc) is abnormally deposited in the extracellular space of diseased CNS. Neurodegeneration in these disease has been shown to be associated with accumulation of PrPSc in affected tissue. To investigate the possible uptake mechanisms that may be required for PrPSc-induced neurodegeneration we studied the cellular trafficking of the neurotoxic fragment, PrP106-126. We were able to detect, by fluorescence microscopy, PrP106-126 inclusions in murine neurones, astrocytes and microglia in vitro. These inclusions were abundant after 24 hour exposure and still present 48h post-exposure. Shorter exposure times yielded only occasional cells with inclusions. Large extracellular aggregates of PrP106-126 could also be detected, which appeared in a time dependent manner. The appearance of inclusions or aggregates was not dependent on PrPc expression as determined by exposure of peptides from PrP-null mice. Using transmission electron microscopy and gold particle detection, positively labelled osmiophilic inclusions of peptide could be detected in the cytoplasm of exposed cells. These results demonstrate that cultured cells are capable of sequestering PrP106-126 and may indicate uptake pathways for PrPSc in various cell types. Toxicity of PrP106-126 may thus be mediated via a sequestration pathway that is not effective for this peptide in PrP-null cells.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1007028323666
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