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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Journal of neurocytology 9 (1980), S. 305-320 
    ISSN: 1573-7381
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary We have developed a combined staining technique whereby pre- and postsynaptic structures of the neuromuscular junction can be simultaneously visualized in the light microscope. The general approach was first to stain presynaptic nerve terminals using nitroblue tetrazolium (NBT), which when reduced to its diformazan state coloured the entire nerve terminal arborization blue. When the NBT stain was combined with the Karnovsky acetylcholinesterase (AChE) procedure the blue-coloured nerve terminal processes were vividly outlined by the brown copper ferrocyanide-AChE reaction product. Experiments were performed to ensure that the NBT-AChE method stained nerve reliably and that even extremely small neural processes were stained and visualized in the light microscope. This method stained cholinergic neuromuscular junctions and unmyelinated axons in a variety of preparations. Aldehyde pre- and poststaining fixation markedly affected the quality of NBT nerve stain. In addition, glutaraldehyde had a direct role in the staining process. The quality of the nerve terminal staining was affected by the pH and the constituents of the staining solution. The powerful experimental advantage of the NBT-AChE stain for neuromuscular junctions resulted from the sharp colour contrast which made possible accurate determinations of the relationship between the presynaptic nerve terminal arborization and the postsynaptic junctional AChE activity.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Journal of neurocytology 5 (1976), S. 691-718 
    ISSN: 1573-7381
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Regenerating neuromuscular junctions in the cutaneous pectoris muscle of the frog were examined by light and electron microscopy up to three months after crushing the motor nerve. The aim was to determine the precision of reinnervation of the original synaptic sites. More than 95% of the original postsynaptic membrane is recovered by nerve terminals and little, if any, synaptic contact is made on other portions of the muscle fibre surface. Even after prolonged denervation when the Schwann cells have retracted from 70–80% of the postsynaptic membrane, regenerating terminals return to and cover a large fraction of it. Although synapses are confined to the original synaptic sites, the pattern of innervation of muscle fibres is altered in several ways: (a) regenerating axon terminals can fail to branch leaving small stretches of postsynaptic membrane uncovered; (b) two terminal branches can lie side by side over a stretch of postsynaptic membrane normally occupied by one terminal; and (c) after growing along a stretch of postsynaptic membrane on one muscle fibre, terminals can leave it to end either in extracellular space or on the postsynaptic membrane of another fibre. Altogether the results demonstrate a strong and specific affinity between the original synaptic sites and regenerating nerve terminals.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Journal of neurocytology 9 (1980), S. 321-342 
    ISSN: 1573-7381
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Developing neuromuscular junctions in the cutaneous pectoris muscle from tadpoles and postmetamorphic frogs were studied in the light microscope. Presynaptic nerve terminals and postsynaptic acetylcholinesterase (AChE) activity were simultaneously demonstrated at these developing junctions using the NBT-AChE method. The earliest nerve contacts studied were small enlargements at the ends of unmyelinated axons. As development progressed, single nerve contacts, often with growth cones, grew in length, generally parallel to the long axis of the myotube. Further endplate maturation was characterized by individual terminal processes developing secondary and tertiary branches, ultimately leading to highly complex terminal arborizations. The initial synaptic contact at developing neuromuscular junctions was made by a single axon, but with further development these same synaptic sites became multiply innervated. The occurrence of multiple innervation was a transient phenomena; the multiple synaptic inputs were eliminated during further maturation. The time course of synapse elimination was protracted, with some multiple innervation even persisting in relatively large adult frogs. The first nerve contacts were generally devoid of observable endplate AChE activity. Early appearance of AChE activity was sometimes graded and in some cases portions of the nerve terminal processes were associated with AChE while other regions of the same terminal arborization were not.
    Type of Medium: Electronic Resource
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