Depolarizing Agents and Cyclic Nucleotides Regulate the Phosphorylation of Specific Neuronal Proteins in Rat Cerebral Cortex Slices

The regulation of the state of phosphorylation of two specific neuronal proteins, designated protein Ia and protein Ib, has been studied in slices of rat cerebral cortex incubated in vitro. For this purpose, a method was developed that prevents dephosphorylation of these proteins during their extrac...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1978-10, Vol.75 (10), p.5195-5199
Main Authors: Forn, Javier, Greengard, Paul
Format: Article
Language:English
Subjects:
1-Methyl-3-isobutylxanthine - pharmacology
Animals
Brain
Calcium
Calcium - metabolism
Cerebral cortex
Cerebral Cortex - metabolism
Cyclic AMP - physiology
Cyclic nucleotides
Gels
Homogenization
In Vitro Techniques
Nerve Tissue Proteins - metabolism
Neurons
Phosphatases
Phosphorylation
Potassium - pharmacology
Rats
Tetrodotoxin
Tetrodotoxin - pharmacology
Veratridine - pharmacology
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Summary:The regulation of the state of phosphorylation of two specific neuronal proteins, designated protein Ia and protein Ib, has been studied in slices of rat cerebral cortex incubated in vitro. For this purpose, a method was developed that prevents dephosphorylation of these proteins during their extraction. When the slices were incubated in a standard Krebs-Ringer solution, proteins Ia and Ib were present almost entirely in the dephosphorylated form. Incubation with cyclic AMP, 8-bromo cyclic AMP, N6-monobutyryl cyclic AMP, or with a phosphodiesterase inhibitor, 3-isobutyl-1-methylxanthine, increased the phosphorylation of proteins Ia and Ib in the slices. Depolarization of neuronal membranes by high K+ or by veratridine was also associated with an increased phosphorylation of proteins Ia and Ib. The effect of depolarizing agents, but not that of cyclic nucleotides or 3-isobutyl-1-methylxanthine, required the presence of external Ca2+ in the incubation medium. Tetrodotoxin blocked the stimulation of the phosphorylation of proteins Ia and Ib induced by veratridine but not that induced by the other agents tested. Incubation of the brain slices with 8-bromo cyclic AMP, 3-isobutyl-1-methylxanthine, high K+, or veratridine also increased the state of phosphorylation of two other neuronal proteins found in extracts of the slices.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.75.10.5195