Regulation of presynaptic strength by controlling Ca2+ channel mobility: effects of cholesterol depletion on release at the cone ribbon synapse

J Neurophysiol. 2012 Jun;107(12):3468-78. doi: 10.1152/jn.00779.2011. Epub 2012 Mar 21.


Synaptic communication requires proper coupling between voltage-gated Ca(2+) (Ca(V)) channels and synaptic vesicles. In photoreceptors, L-type Ca(V) channels are clustered close to synaptic ribbon release sites. Although clustered, Ca(V) channels move continuously within a confined domain slightly larger than the base of the ribbon. We hypothesized that expanding Ca(V) channel confinement domains should increase the number of channel openings needed to trigger vesicle release. Using single-particle tracking techniques, we measured the expansion of Ca(V) channel confinement domains caused by depletion of membrane cholesterol with cholesterol oxidase or methyl-β-cyclodextrin. With paired whole cell recordings from cones and horizontal cells, we then determined the number of Ca(V) channel openings contributing to cone Ca(V) currents (I(Ca)) and the number of vesicle fusion events contributing to horizontal cell excitatory postsynaptic currents (EPSCs) following cholesterol depletion. Expansion of Ca(V) channel confinement domains reduced the peak efficiency of release, decreasing the number of vesicle fusion events accompanying opening of each Ca(V) channel. Cholesterol depletion also inhibited exocytotic capacitance increases evoked by brief depolarizing steps. Changes in efficiency were not due to changes in I(Ca) amplitude or glutamate receptor properties. Replenishing cholesterol restored Ca(V) channel domain size and release efficiency to control levels. These results indicate that cholesterol is important for organizing the cone active zone. Furthermore, the finding that cholesterol depletion impairs coupling between channel opening and vesicle release by allowing Ca(V) channels to move further from release sites shows that changes in presynaptic Ca(V) channel mobility can be a mechanism for adjusting synaptic strength.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Ambystoma
  • Animals
  • Calcium Channels, L-Type / drug effects
  • Calcium Channels, L-Type / physiology*
  • Cholesterol / physiology*
  • Cholesterol Oxidase / pharmacology
  • Excitatory Postsynaptic Potentials / drug effects
  • Excitatory Postsynaptic Potentials / physiology
  • Female
  • Male
  • Patch-Clamp Techniques
  • Receptors, Glutamate / physiology
  • Retinal Cone Photoreceptor Cells / drug effects
  • Retinal Cone Photoreceptor Cells / physiology*
  • Retinal Horizontal Cells / drug effects
  • Retinal Horizontal Cells / physiology
  • Synapses / drug effects
  • Synapses / physiology
  • Synaptic Vesicles / drug effects
  • Synaptic Vesicles / physiology
  • beta-Cyclodextrins / pharmacology


  • Calcium Channels, L-Type
  • Receptors, Glutamate
  • beta-Cyclodextrins
  • methyl-beta-cyclodextrin
  • Cholesterol
  • Cholesterol Oxidase