Mutant PKCγ in spinocerebellar ataxia type 14 disrupts synapse elimination and long-term depression in Purkinje cells in vivo

J Neurosci. 2011 Oct 5;31(40):14324-34. doi: 10.1523/JNEUROSCI.5530-10.2011.


Cerebellar Purkinje cells (PCs) express a large amount of the γ isoform of protein kinase C (PKCγ) and a modest level of PKCα. The PKCγ is involved in the pruning of climbing fiber (CF) synapses from developing PCs, and PKCα plays a critical role in long-term depression (LTD) at parallel fiber (PF)-PC synapses. Moreover, the PKC signaling in PCs negatively modulates the nonselective transient receptor potential cation channel type 3 (TRPC3), the opening of which elicits slow EPSCs at PF-PC synapses. Autosomal dominant spinocerebellar ataxia type 14 (SCA14) is caused by mutations in PKCγ. To clarify the pathology of this disorder, mutant (S119P) PKCγ tagged with GFP was lentivirally expressed in developing and mature mouse PCs in vivo, and the effects were assessed 3 weeks after the injection. Mutant PKCγ-GFP aggregated in PCs without signs of degeneration. Electrophysiology results showed impaired pruning of CF synapses from developing PCs, failure of LTD expression, and increases in slow EPSC amplitude. We also found that mutant PKCγ colocalized with wild-type PKCγ, which suggests that mutant PKCγ acts in a dominant-negative manner on wild-type PKCγ. In contrast, PKCα did not colocalize with mutant PKCγ. The membrane residence time of PKCα after depolarization-induced translocation, however, was significantly decreased when it was present with the mutant PKCγ construct. These results suggest that mutant PKCγ in PCs of SCA14 patients could differentially impair the membrane translocation kinetics of wild-type γ and α PKCs, which would disrupt synapse pruning, synaptic plasticity, and synaptic transmission.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Membrane / enzymology
  • Cell Membrane / genetics
  • Cells, Cultured
  • Cerebellum / enzymology
  • Female
  • Humans
  • Isoenzymes / genetics
  • Isoenzymes / metabolism
  • Long-Term Synaptic Depression / genetics*
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Mutation / physiology*
  • Organ Culture Techniques
  • Protein Kinase C / biosynthesis
  • Protein Kinase C / genetics*
  • Protein Kinase C / metabolism
  • Protein Kinase C-alpha / genetics
  • Protein Kinase C-alpha / metabolism
  • Protein Transport / genetics
  • Purkinje Cells / enzymology*
  • Spinocerebellar Ataxias
  • Spinocerebellar Degenerations / enzymology*
  • Spinocerebellar Degenerations / genetics
  • Synapses / enzymology*
  • Synapses / genetics


  • Isoenzymes
  • protein kinase C gamma
  • Protein Kinase C
  • Protein Kinase C-alpha

Supplementary concepts

  • Spinocerebellar ataxia 14