Opposing changes in phosphorylation of specific sites in synapsin I during Ca2+-dependent glutamate release in isolated nerve terminals

J Neurosci. 2001 Oct 15;21(20):7944-53. doi: 10.1523/JNEUROSCI.21-20-07944.2001.


Synapsins are major neuronal phosphoproteins involved in regulation of neurotransmitter release. Synapsins are well established targets for multiple protein kinases within the nerve terminal, yet little is known about dephosphorylation processes involved in regulation of synapsin function. Here, we observed a reciprocal relationship in the phosphorylation-dephosphorylation of the established phosphorylation sites on synapsin I. We demonstrate that, in vitro, phosphorylation sites 1, 2, and 3 of synapsin I (P-site 1 phosphorylated by cAMP-dependent protein kinase; P-sites 2 and 3 phosphorylated by Ca(2+)-calmodulin-dependent protein kinase II) were excellent substrates for protein phosphatase 2A, whereas P-sites 4, 5, and 6 (phosphorylated by mitogen-activated protein kinase) were efficiently dephosphorylated only by Ca(2+)-calmodulin-dependent protein phosphatase 2B-calcineurin. In isolated nerve terminals, rapid changes in synapsin I phosphorylation were observed after Ca(2+) entry, namely, a Ca(2+)-dependent phosphorylation of P-sites 1, 2, and 3 and a Ca(2+)-dependent dephosphorylation of P-sites 4, 5, and 6. Inhibition of calcineurin activity by cyclosporin A resulted in a complete block of Ca(2+)-dependent dephosphorylation of P-sites 4, 5, and 6 and correlated with a prominent increase in ionomycin-evoked glutamate release. These two opposing, rapid, Ca(2+)-dependent processes may play a crucial role in the modulation of synaptic vesicle trafficking within the presynaptic terminal.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • 4-Aminopyridine / pharmacology
  • Animals
  • Binding Sites / physiology
  • Calcium / metabolism*
  • Cerebral Cortex / chemistry
  • Enzyme Inhibitors / pharmacology
  • Glutamic Acid / metabolism*
  • Kinetics
  • Male
  • Phosphoprotein Phosphatases / antagonists & inhibitors
  • Phosphoprotein Phosphatases / chemistry
  • Phosphoprotein Phosphatases / metabolism
  • Phosphorylation / drug effects
  • Presynaptic Terminals / metabolism*
  • Protein Phosphatase 2
  • Rats
  • Rats, Sprague-Dawley
  • Substrate Specificity / physiology
  • Synapsins / chemistry
  • Synapsins / metabolism*
  • Synaptosomes / chemistry
  • Synaptosomes / metabolism*


  • Enzyme Inhibitors
  • Synapsins
  • Glutamic Acid
  • 4-Aminopyridine
  • Phosphoprotein Phosphatases
  • Protein Phosphatase 2
  • Calcium