Mutant NG108-15 cells (NG-CR72) deficient in GM1 synthase respond aberrantly to axonogenic stimuli and are vulnerable to calcium-induced apoptosis: they are rescued with LIGA-20.

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  • Author(s): Wu G;Wu G; Lu ZH; Xie X; Li L; Ledeen RW
  • Source:
    Journal of neurochemistry [J Neurochem] 2001 Feb; Vol. 76 (3), pp. 690-702.
  • Publication Type:
    Journal Article; Research Support, U.S. Gov't, P.H.S.
  • Language:
    English
  • Additional Information
    • Source:
      Publisher: Wiley on behalf of the International Society for Neurochemistry Country of Publication: England NLM ID: 2985190R Publication Model: Print Cited Medium: Print ISSN: 0022-3042 (Print) Linking ISSN: 00223042 NLM ISO Abbreviation: J Neurochem Subsets: MEDLINE
    • Publication Information:
      Publication: 2001- : Oxford, UK : Wiley on behalf of the International Society for Neurochemistry
      Original Publication: New York : Raven Press
    • Subject Terms:
      Apoptosis*; Axons/*physiology ; Calcium/*pharmacology ; G(M1) Ganglioside/*analogs & derivatives ; Galactosyltransferases/*physiology ; Sphingosine/*analogs & derivatives; Bucladesine/pharmacology ; G(M1) Ganglioside/pharmacology ; Galactosyltransferases/deficiency ; Galactosyltransferases/genetics ; Ganglioside Galactosyltransferase ; Hybrid Cells ; Ionomycin/pharmacology ; Ionophores/pharmacology ; Mutation ; Neuroprotective Agents/pharmacology ; Potassium Chloride/pharmacology ; Sphingosine/pharmacology ; Tretinoin/pharmacology ; Tumor Cells, Cultured/drug effects
    • Abstract:
      The neuroblastoma x glioma NG108-15 hybrid cell line, a widely used model for the study of neuronal differentiation, contains a variety of gangliosides including GM1 and its sialosylated derivative, GD1a. To investigate the role of these a-series gangliotetraose gangliosides in neuritogenesis, we have obtained a mutated subclone of NG108-15 that is deficient in that family of gangliosides. NG108-15 cells were grown in the presence of cholera toxin, which killed the large majority of cells, and from the cholera-resistant survivors we isolated a clone, NG-CR72, that lacks GM1 and GD1a in the plasma and nuclear membranes. GM2 concentration was significantly higher in the plasma membrane. Enzyme assay indicated deficiency of UDP-Gal:GM2 galactosyltransferase (GM1 synthase), which was confirmed by incorporation studies with [3H]sphingosine. These cells resembled wild-type NG108-15 in extending dendritic processes in response to dendritogenic agents (retinoic acid, dibutyryl cAMP) but responded aberrantly to axonogenic stimuli (KCl, ionomycin) by extending unstable neurites that showed the cytoskeletal staining characteristic of dendrites. Moreover, mutant cells treated with the Ca2+ elevating axonogenic agents underwent apoptosis over time, attributed to dysfunction of Ca2+ regulatory mechanisms normally mediated by GM1. Such agents caused dramatic and sustained elevation of intracellular Ca2+ in mutant cells, in contrast to modest and temporary elevation in wild-type cells. Exogenous GM1, inserted into the plasma membrane, had no discernable protective effect on NG-CR72 cells whereas LIGA-20, a membrane-permeant derivative of GM1 that entered both plasma and nuclear membranes, blocked apoptosis, permitted extension of stable neurites, and attenuated the abnormal elevation of intracellular Ca2+.
    • Grant Information:
      NS33912 United States NS NINDS NIH HHS
    • Accession Number:
      0 (Ionophores)
      0 (Neuroprotective Agents)
      126586-85-4 (LIGA 20)
      37758-47-7 (G(M1) Ganglioside)
      56092-81-0 (Ionomycin)
      5688UTC01R (Tretinoin)
      63X7MBT2LQ (Bucladesine)
      660YQ98I10 (Potassium Chloride)
      EC 2.4.1.- (Galactosyltransferases)
      EC 2.4.1.62 (Ganglioside Galactosyltransferase)
      NGZ37HRE42 (Sphingosine)
      SY7Q814VUP (Calcium)
    • Publication Date:
      Date Created: 20010207 Date Completed: 20010315 Latest Revision: 20190630
    • Publication Date:
      20240829
    • Accession Number:
      10.1046/j.1471-4159.2001.00036.x
    • Accession Number:
      11158239