Summary
Organotypic cultures of mammalian embryonic spinal cord-dorsal root ganglia combinations treated with H3 thymidine, were studied with light and electron microscopic autoradiography. Cultures pulsed with H3 thymidine for up to six hours showed 0–2% of labelled neuroepithelial cells when fixed immediately thereafter, and 7 to 15 times as many labelled cells when fixed on subsequent daysin vitro (DIV). Cultures exposed to radioactive material for days, at the initiation of the culture, and prior to myelinationin vitro, were compared. The cumulative evidence showed that the peak of proliferative activity occurred not after explantation, but between DIV 7 and DIV 12, which is after synapse formation and prior to myelination. Another smaller proliferative peak occurred after myelination between DIV 15 and DIV 19.
Morphological observations with the light microscope revealed labelling of predominantly small dark cells during the first proliferative peak, that is, prior to myelination. Using electron microscopic criteria for identification, these small dark cells were “large glioblasts”, “large glial precursors” and “young” oligodendrocytes, and transitions could be observed between these cells in that order of differentiation. Oligodendrocytes when closely connected with myelin sheaths did not become labelled.
Labelling and proliferation of medium large light cells inconspicuously preceded that of oligodendrocytes and their precursors, and continued modestly throughout myelination, achieving a modest peak and predominance during DIV 15 and DIV 19. Astrocytic features could be demonstrated ultrastructurally in these medium large light cells and mitotic division in this type of cell was observed. The possibility that part of this astrocytic population arose from “large glioblasts” was discussed.
No labelled cells were seen ultrastructurally fulfilling criteria of amitotic division.
No neurons showed labelling with H3 thymidine.
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Supported by a USPHS Grant No. NB-07849-03.
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Manuelidis, L., Manuelidis, E.E. An autoradiographic study of the proliferation and differentiation of glial cellsin vitro . Acta Neuropathol 18, 193–213 (1971). https://doi.org/10.1007/BF00685066
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DOI: https://doi.org/10.1007/BF00685066