3
views
0
recommends
+1 Recommend
0 collections
    0
    shares
      • Record: found
      • Abstract: found
      • Article: not found

      Investigating the feasibility of channelrhodopsin variants for nanoscale optogenetics.

      Read this article at

      ScienceOpenPublisherPMC
      Bookmark
          There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.

          Abstract

          Optogenetics has revolutionized the study of circuit function in the brain, by allowing activation of specific ensembles of neurons by light. However, this technique has not yet been exploited extensively at the subcellular level. Here, we test the feasibility of a focal stimulation approach using stimulated emission depletion/reversible saturable optical fluorescence transitions-like illumination, whereby switchable light-gated channels are focally activated by a laser beam of one wavelength and deactivated by an overlapping donut-shaped beam of a different wavelength, confining activation to a center focal region. This method requires that activated channelrhodopsins are inactivated by overlapping illumination of a distinct wavelength and that photocurrents are large enough to be detected at the nanoscale. In tests of current optogenetic tools, we found that ChR2 C128A/H134R/T159C and CoChR C108S and C108S/D136A-activated with 405-nm light and inactivated by coillumination with 594-nm light-and C1V1 E122T/C167S-activated by 561-nm light and inactivated by 405-nm light-were most promising in terms of highest photocurrents and efficient inactivation with coillumination. Although further engineering of step-function channelrhodopsin variants with higher photoconductances will be required to employ this approach at the nanoscale, our findings provide a framework to guide future development of this technique.

          Related collections

          Author and article information

          Journal
          Neurophotonics
          Neurophotonics
          SPIE-Intl Soc Optical Eng
          2329-423X
          2329-423X
          Jan 2019
          : 6
          : 1
          Affiliations
          [1 ] European Neuroscience Institute, Trans-Synaptic Signaling Group, Goettingen, Germany.
          [2 ] Stanford University, Howard Hughes Medical Institute, Department of Bioengineering, Department of Psychiatry, CNC Program, Stanford, California, United States.
          [3 ] University Medical Center, Center for Nanoscale Microscopy and Molecular Physiology of the Brain, Goettingen, Germany.
          [4 ] MIT Media Lab and McGovern Institute, Departments of Brain and Cognitive Science and Biological Engineering, Cambridge, Massachusetts, United States.
          Article
          18062RR
          10.1117/1.NPh.6.1.015007
          6393647
          30854405
          09c3ee5c-f8b3-46cc-9206-abaa7e479dba
          History

          RESOLFT microscopy,STED microscopy,channelrhodopsin,optogenetics

          Comments

          Comment on this article