Biochemical and electrophysiological differentiation profile of a human neuroblastoma (IMR-32) cell line.
A human neuroblastoma cell line (IMR-32), when differentiated, mimics large projections of the human cerebral cortex and under certain tissue culture conditions, forms intracellular fibrillary material, commonly observed in brains of patients affected with Alzheimer's disease. Our purpose is to use differentiated IMR-32 cells as an in vitro system for magnetic field exposure studies. We have previously studied in vitro differentiation of murine neuroblastoma (N1E-115) cells with respect to resting membrane potential development. The purpose of this study was to extend our investigation to IMR-32 cells. Electrophysiological (resting membrane potential, V(m)) and biochemical (neuron-specific enolase activity [NSE]) measurements were taken every 2 d for a period of 16 d. A voltage-sensitive oxonol dye together with flow cytometry was used to measure relative changes in V(m). To rule out any effect due to mechanical cell detachment, V(m) was indirectly measured by using a slow potentiometric dye (tetramethylrhodamine methyl ester) together with confocal digital imaging microscopy. Neuron-specific enolase activity was measured by following the production of phosphoenolpyruvate from 2-phospho-d-glycerate at 240 nm. Our results indicate that in IMR-32, in vitro differentiation as characterized by an increase in NSE activity is not accompanied by resting membrane potential development. This finding suggests that pathways for morphological-biochemical and electrophysiological differentiations in IMR-32 cells are independent of one another.
Rao
Raj R
RR
Cellular Bioengineering Laboratory, Biological and Agricultural Engineering Department, Driftmier Engineering Center, University of Georgia, Athens 30602, USA.
Kisaalita
William S
WS
D002454:Cell Differentiation; D005434:Flow Cytometry; D005453:Fluorescence; D006801:Humans; D009447:Neuroblastoma / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D010751:Phosphopyruvate Hydratase / Q000378:metabolism; D014407:Tumor Cells, Cultured
D010751:Phosphopyruvate Hydratase