Protein kinase C and calmodulin serve as calcium sensors for calcium-stimulated endocytosis at synapses

Ying Hui Jin, Xin Sheng Wu, Bo Shi, Zhen Zhang, Xiaoli Guo, Lin Gan, Zhongqing Chen, Ling Gang Wu

Research output: Contribution to journalArticle

2 Scopus citations

Abstract

Calcium influx triggers and facilitates endocytosis, which recycles vesicles and thus sustains synaptic transmission. Despite decades of studies, the underlying calcium sensor remained not well understood. Here, we examined two calcium binding proteins, protein kinase C (PKC) and calmodulin. Whether PKC is involved in endocytosis was unclear; whether calmodulin acts as a calcium sensor for endocytosis was neither clear, although calmodulin involvement in endocytosis had been suggested. We generated PKC (α or β-isoform) and calmodulin (calmodulin 2 gene) knock-out mice of either sex and measured endocytosis with capacitance measurements, pHluorin imaging and electron microscopy. We found that these knock-outs inhibited slow (~10 –30 s) and rapid (<~3 s) endocytosis at large calyx-type calyces, and inhibited slow endocytosis and bulk endocytosis (forming large endosome-like structures) at small conventional hippocampal synapses, suggesting the involvement of PKC and calmodulin in three most common forms of endocytosis-the slow, rapid and bulk endocytosis. Inhibition of slow endocytosis in PKC or calmodulin 2 knock-out hippocampal synapses was rescued by overexpressing wild-type PKC or calmodulin, but not calcium-binding-deficient PKC or calmodulin mutant, respectively, suggesting that calcium stimulates endocytosis by binding with its calcium sensor PKC and calmodulin. PKC and calmodulin 2 knock-out inhibited calcium-dependent vesicle mobilization to the readily releasable pool, suggesting that PKC and calmodulin may mediate calcium-dependent facilitation of vesicle mobilization. These findings shed light on the molecular signaling link among calcium, endocytosis and vesicle mobilization that are crucial in maintaining synaptic transmission and neuronal network activity.

Original languageEnglish (US)
Pages (from-to)9478-9490
Number of pages13
JournalJournal of Neuroscience
Volume39
Issue number48
DOIs
StatePublished - Nov 27 2019

Keywords

  • Calmodulin
  • Capacitance measurement
  • Electron microscopy
  • Endocytosis
  • PHluorin imaging
  • Protein kinase C

ASJC Scopus subject areas

  • Neuroscience(all)

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