Induced and spontaneous neuritogenesis are associated with enhanced expression of ganglioside GM1 in the nuclear membrane.
Neuro-2a neuroblastoma cells can be stimulated to extend neurites with a number of agents, one of which, neuraminidase, induces terminal differentiation by a mechanism involving enhanced Ca2+ influx. Permeabilization of such differentiated cells with saponin and treatment with cholera toxin B subunit linked to horseradish peroxidase revealed intense staining of the nuclear membrane, indicating the presence of GM1 ganglioside. Unstimulated cells had barely detectable levels of nuclear GM1. Nuclei isolated by sucrose density gradient centrifugation similarly showed intense staining with fluorescently labeled cholera toxin B subunit, in contrast to nuclei from undifferentiated controls. Treatment with chloroform-methanol removed most of the fluorogenic material. Chemical analysis of such nuclei from neuraminidase-treated cells confirmed significant elevation of GM1 above control levels, along with virtual absence of markers for plasma membrane and Golgi apparatus. Cerebellar granule cells from neonatal rats revealed a similar phenomenon following spontaneous neurite outgrowth in culture.
Wu
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New Jersey Medical School, UMDNJ, Department of Neurosciences MSB-H506, Newark 07103, USA.
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Ledeen
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D000818:Animals; D002118:Calcium / Q000378:metabolism; D002454:Cell Differentiation; D002460:Cell Line; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D002531:Cerebellum / Q000166:cytology / Q000502:physiology*; D002772:Cholera Toxin / Q000494:pharmacology; D005677:G(M1) Ganglioside / Q000096:biosynthesis*; D006735:Horseradish Peroxidase / Q000494:pharmacology; D008563:Membrane Lipids / Q000096:biosynthesis; D051379:Mice; D009439:Neuraminidase / Q000494:pharmacology; D016501:Neurites / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D009447:Neuroblastoma; D009474:Neurons / Q000187:drug effects / Q000502:physiology / Q000648:ultrastructure; D009685:Nuclear Envelope / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012503:Saponins / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D008563:Membrane Lipids; D012503:Saponins; C052635:cholera toxin, B subunit-horseradish peroxidase; D005677:G(M1) Ganglioside; D002772:Cholera Toxin; D006735:Horseradish Peroxidase; D009439:Neuraminidase; D002118:Calcium