Adrenergic and calcium-mediated subcellular redistribution of protein kinase C in primary neuronal cultures

• Published in: Biochemical and biophysical research communications Vol. 166; no. 1; pp. 22 - 28
• Main Authors: Kalberg, Christopher J., Sumners, Colin
• Format: Journal Article
• Language: English
• Published: San Diego, CA Elsevier Inc 15.01.1990; Elsevier
• Subjects: Analytical, structural and metabolic biochemistry; Animals; Animals, Newborn; Biological and medical sciences; Brain - enzymology; Calcium Chloride - pharmacology; Cell Fractionation - methods; Cells, Cultured; Cytosol - enzymology; Egtazic Acid - pharmacology; Enzymes and enzyme inhibitors; Epinephrine - pharmacology; Fundamental and applied biological sciences. Psychology; Kinetics; Neurons - drug effects; Neurons - enzymology; Neurons - ultrastructure; Prazosin - pharmacology; Protein Kinase C - metabolism; Rats; Rats, Inbred Strains; Tetradecanoylphorbol Acetate - pharmacology; Transferases; DMEM; DG; TPA; PMSF; DEAE; PDHS; Ang II; Epinephrine; Cell culture; Neonatal; Protein kinase C; Radiolabelling; Rat; Enzyme; Rodentia; α»-Adrenergic receptor agonist; Activation; Calcium Ions; Vertebrata; Mammalia; Neuron; Enzymatic activity; α-Adrenergic receptor; Column chromatography
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/0006-291X(90)91906-9

Incubation of primary neuronal cultures prepared from the brains of neonatal rats with 50 μM epinephrine resulted in the transient redistribution of protein kinase C from the cytosol to the particulate fraction. This effect occurred after 1 and 5 min of incubation and resulted in a decrease in cytosolic protein kinase C activity with a corresponding increase in particulate protein kinase C of approximately 30% and 15%, respectively. The epinephrine-stimulated translocation of protein kinase C was blocked by 1 μM prazosin indicating the involvement of α 1-adrenergic receptors. Further, inclusion of 0.1 μM Ca 2+ in the homogenization buffer was found to significantly enhance the binding of protein kinase C to cellular membranes prepared from neuronal cultures. These results indicate that α 1-adrenergic receptors in neuronal brain cell cultures are linked to the activation of protein kinase C and that the mobilization of Ca 2+ may enhance this effect.