Premium
Inositol Metabolism During Neuroblastoma B50 Cell Differentiation: Effects of Differentiating Agents on Inositol Uptake
Author(s) -
Reboulleau Christian P.
Publication year - 1990
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1990.tb04181.x
Subject(s) - extracellular , inositol , ouabain , biochemistry , intracellular , phloretin , biology , metabolism , calcium , chemistry , biophysics , sodium , receptor , organic chemistry
Inositol uptake was studied in the rat CNS neuroblastoma B50 cell line. Eadie‐Hofstee analysis of the uptake pattern reveals two defined modes of inositol entry into the cell. The high‐affinity uptake component requires the presence of extracellular sodium and is inhibited by phloridzin. Analysis of the uptake velocities of the high‐affinity uptake component provided the following apparent kinetic parameters: K m = 13.7 μ M and V max = 14.7 pmol/mg of protein/min (without correcting for residual diffusion) and K m = 12.9 μ M and V max = 12.3 pmol/mg of protein/min (with correction). At physiological concentrations, the high‐affinity . transport process contributes approximately 70% to total uptake; the remainder is due to a low‐affinity diffusion‐like process. Uptake inhibition studies reveal that the uptake process is sensitive to ouabain, amiloride, and dichlorobenzamil inhibition but relatively insensitive to cytochalasin B or phloretin. When neuroblastoma B50 cells are induced to differentiate morphologically with high extracellular calcium or with dibutyryl cyclic AMP, a significant decrease in inositol uptake is observed. The dibutyrl cyclic AMP‐mediated inhibition of uptake affects only the high‐affinity uptake component and is noncompetitive in nature. The high extracellular calcium‐mediated inhibition is less specific; it involves “disappearance” of the high‐affinity process, some inhibition of the low‐affinity process, and an increase of inositol efflux. The significance of these observations is discussed in the context of neuroblastoma B50 cell differentiation.