title: Dynamics of Phosphoinositide-Dependent Signaling in Sympathetic Neurons.
abstract: In neurons, loss of plasma membrane phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] leads to a decrease in exocytosis and changes in electrical excitability. Restoration of PI(4,5)P2 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)P2, phosphatidylinositol 4-phosphate, diacylglycerol, inositol 1,4,5-trisphosphate, and Ca(2+) 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)P2 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)P2-dependent processes in neurons.
  
  
  CONCLUSIONS
  Phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] is a minor phospholipid in the cytoplasmic leaflet of the plasma membrane. Depletion of PI(4,5)P2 via phospholipase C-mediated hydrolysis leads to a decrease in exocytosis and alters electrical excitability in neurons. Restoration of PI(4,5)P2 is essential for a return to basal neuronal activity. However, the dynamics of phosphoinositide metabolism have not been analyzed in neurons. We studied the dynamics of phosphoinositide metabolism in sympathetic neurons upon muscarinic stimulation and used the kinetic information to develop a quantitative description of neuronal phosphoinositide metabolism. The measurements and analysis show a several-fold faster synthesis of PI(4,5)P2 in sympathetic neurons than in an electrically nonexcitable cell line, and provide a framework for future studies of PI(4,5)P2-dependent processes in neurons.
authors.affiliation: Department of Physiology and Biophysics, University of Washington, Seattle, Washington 98195, and.
authors.forename: Martin
authors.identifier: http://orcid.org/0000-0003-4301-8385
authors.initials: M
authors.lastname: Kruse
authors.affiliation: Department of Physiology and Biophysics, University of Washington, Seattle, Washington 98195, and.
authors.forename: Oscar
authors.initials: O
authors.lastname: Vivas
authors.affiliation: ATK Innovation, Analytics and Discovery, North Bend, Washington 98045.
authors.forename: Alexis
authors.initials: A
authors.lastname: Traynor-Kaplan
authors.affiliation: Department of Physiology and Biophysics, University of Washington, Seattle, Washington 98195, and hille@u.washington.edu.
authors.forename: Bertil
authors.identifier: http://orcid.org/0000-0002-7266-1671
authors.initials: B
authors.lastname: Hille
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D006801:Humans; D051658:KCNQ2 Potassium Channel / Q000235:genetics / Q000378:metabolism; D051659:KCNQ3 Potassium Channel / Q000235:genetics / Q000378:metabolism; D008297:Male; D008564:Membrane Potentials / Q000235:genetics; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D009474:Neurons / Q000502:physiology*; D017711:Nonlinear Dynamics; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000378:metabolism; D010716:Phosphatidylinositols / Q000235:genetics / Q000378:metabolism*; D054803:Phospholipase C delta / Q000235:genetics / Q000378:metabolism; D011506:Proteins / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D043643:Receptor, Muscarinic M1 / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology*; D017783:Superior Cervical Ganglion / Q000166:cytology*; D013997:Time Factors
chemical_list: D051658:KCNQ2 Potassium Channel; D051659:KCNQ3 Potassium Channel; D008565:Membrane Proteins; D019269:Phosphatidylinositol 4,5-Diphosphate; D010716:Phosphatidylinositols; D011506:Proteins; C109563:RTL8C protein, human; D043643:Receptor, Muscarinic M1; C546203:TUB protein, rat; C518024:PLCD1 protein, human; D054803:Phospholipase C delta; D002118:Calcium
keywords: M-current; PI(4,5)P2; excitability; phosphoinositide metabolism; superior cervical ganglion neurons
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