Dynamics of Phosphoinositide-Dependent Signaling in Sympathetic Neurons

In neurons, loss of plasma membrane phosphatidylinositol 4,5-bisphosphate [PI(4,5)P-2] leads to a decrease in exocytosis and changes in electrical excitability. Restoration of PI(4,5)P-2 levels after phospholipase C activation is therefore essential for a return to basal neuronal activity. However, the dynamics of phosphoinositide metabolism have not been analyzed in neurons. We measured dynamic changes of PI(4,5)P-2, phosphatidylinositol 4-phosphate, diacylglycerol, inositol 1,4,5-trisphosphate, and Ca2+ upon muscarinic stimulation in sympathetic neurons from adult male Sprague-Dawley rats with electrophysiological and optical approaches. We used this kinetic information to develop a quantitative description of neuronal phosphoinositide metabolism. The measurements and analysis show and explain faster synthesis of PI(4,5)P-2 in sympathetic neurons than in electrically nonexcitable tsA201 cells. They can be used to understand dynamic effects of receptor-mediated phospholipase C activation on excitability and other PI(4,5)P-2-dependent processes in neurons.