Temporally precise single-cell-resolution optogenetics

Nat Neurosci. 2017 Dec;20(12):1796-1806. doi: 10.1038/s41593-017-0018-8. Epub 2017 Nov 13.

Abstract

Optogenetic control of individual neurons with high temporal precision within intact mammalian brain circuitry would enable powerful explorations of how neural circuits operate. Two-photon computer-generated holography enables precise sculpting of light and could in principle enable simultaneous illumination of many neurons in a network, with the requisite temporal precision to simulate accurate neural codes. We designed a high-efficacy soma-targeted opsin, finding that fusing the N-terminal 150 residues of kainate receptor subunit 2 (KA2) to the recently discovered high-photocurrent channelrhodopsin CoChR restricted expression of this opsin primarily to the cell body of mammalian cortical neurons. In combination with two-photon holographic stimulation, we found that this somatic CoChR (soCoChR) enabled photostimulation of individual cells in mouse cortical brain slices with single-cell resolution and <1-ms temporal precision. We used soCoChR to perform connectivity mapping on intact cortical circuits.

MeSH terms

  • Animals
  • Cerebral Cortex / cytology
  • Cerebral Cortex / ultrastructure
  • Imaging, Three-Dimensional
  • Mice
  • Nerve Net / cytology
  • Nerve Net / ultrastructure
  • Neuroimaging / methods*
  • Neurons / physiology*
  • Neurons / ultrastructure
  • Opsins / genetics
  • Optogenetics / instrumentation
  • Optogenetics / methods*
  • Patch-Clamp Techniques
  • Photic Stimulation
  • Receptors, Kainic Acid / genetics
  • Visual Cortex / cytology
  • Visual Cortex / physiology

Substances

  • Gluk2 kainate receptor
  • Opsins
  • Receptors, Kainic Acid