Neuronal and glial epitopes and transmitter-synthesizing enzymes appear in parallel with membrane excitability during neuroblastoma x glioma hybrid differentiation.
The membrane excitability and the presence of neural proteins, including neuronal and glial markers and neurotransmitter-synthesizing enzymes, were examined in parallel while the NG108-15 cell line was maintained in a serum-free medium. Whole-cell recordings in voltage-clamp or current-clamp configurations were used to evaluate the membrane excitability, and immunostaining was done with a panel of well-characterized antibodies against NSE, NF150, S-100 beta, GFAP, ChAT and TH. Culture for 4 to 10 days led to a striking rise in neurite outgrowth, electrical excitability and expression of neural proteins in type I neuron-like cells, which were of both neuronal and glial character, and expressed both cholinergic and adrenergic traits. After about 2 weeks, type II cells which lack neurite processes began to emerge. The type II cells proliferated, as revealed by BrdU uptake, and gradually overgrew differentiated cell types. They exhibited little or no membrane excitability and absence of immunoreactivity for the neuronal and glial specific proteins tested. These measurements indicate that the presence of these neural proteins at crucial stages of membrane excitability development is an important characteristics of NG108-15 cell differentiation, providing insights into the neural development and the reversible nature of neoplasia in the nervous system.
Ma
W
W
Biotechnology Research and Applications Division, Science Applications International, Rockville, MD 20850, USA. wu.ma@cpmx.saic.com
Pancrazio
J J
JJ
Coulombe
M
M
Dumm
J
J
Sathanoori
R
R
Barker
J L
JL
Kowtha
V C
VC
Stenger
D A
DA
Hickman
J J
JJ
D000818:Animals; D002454:Cell Differentiation / Q000502:physiology; D016895:Culture Media, Serum-Free; D000939:Epitopes; D005910:Glioma / Q000201:enzymology / Q000276:immunology*; D006822:Hybrid Cells / Q000276:immunology; D007150:Immunohistochemistry; D008564:Membrane Potentials / Q000502:physiology; D051379:Mice; D009447:Neuroblastoma / Q000201:enzymology / Q000276:immunology*; D009457:Neuroglia / Q000276:immunology*; D009474:Neurons / Q000276:immunology*; D018377:Neurotransmitter Agents / Q000096:biosynthesis*; D018408:Patch-Clamp Techniques; D051381:Rats; D014407:Tumor Cells, Cultured
D016895:Culture Media, Serum-Free; D000939:Epitopes; D018377:Neurotransmitter Agents