Arrays of microLEDs and astrocytes: biological amplifiers to optogenetically modulate neuronal networks reducing light requirement

PLoS One. 2014 Sep 29;9(9):e108689. doi: 10.1371/journal.pone.0108689. eCollection 2014.

Abstract

In the modern view of synaptic transmission, astrocytes are no longer confined to the role of merely supportive cells. Although they do not generate action potentials, they nonetheless exhibit electrical activity and can influence surrounding neurons through gliotransmitter release. In this work, we explored whether optogenetic activation of glial cells could act as an amplification mechanism to optical neural stimulation via gliotransmission to the neural network. We studied the modulation of gliotransmission by selective photo-activation of channelrhodopsin-2 (ChR2) and by means of a matrix of individually addressable super-bright microLEDs (μLEDs) with an excitation peak at 470 nm. We combined Ca2+ imaging techniques and concurrent patch-clamp electrophysiology to obtain subsequent glia/neural activity. First, we tested the μLEDs efficacy in stimulating ChR2-transfected astrocyte. ChR2-induced astrocytic current did not desensitize overtime, and was linearly increased and prolonged by increasing μLED irradiance in terms of intensity and surface illumination. Subsequently, ChR2 astrocytic stimulation by broad-field LED illumination with the same spectral profile, increased both glial cells and neuronal calcium transient frequency and sEPSCs suggesting that few ChR2-transfected astrocytes were able to excite surrounding not-ChR2-transfected astrocytes and neurons. Finally, by using the μLEDs array to selectively light stimulate ChR2 positive astrocytes we were able to increase the synaptic activity of single neurons surrounding it. In conclusion, ChR2-transfected astrocytes and μLEDs system were shown to be an amplifier of synaptic activity in mixed corticalneuronal and glial cells culture.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Astrocytes / drug effects
  • Astrocytes / metabolism*
  • Astrocytes / radiation effects
  • Calcium / metabolism
  • Calcium Signaling / drug effects
  • Calcium Signaling / radiation effects
  • Channelrhodopsins
  • Excitatory Postsynaptic Potentials / drug effects
  • Excitatory Postsynaptic Potentials / radiation effects
  • Female
  • Glutamic Acid / pharmacology
  • Light*
  • Male
  • Nerve Net / drug effects
  • Nerve Net / metabolism*
  • Nerve Net / radiation effects
  • Neuroglia / drug effects
  • Neuroglia / metabolism
  • Neuroglia / radiation effects
  • Neurons / metabolism*
  • Neurons / radiation effects*
  • Optics and Photonics / instrumentation*
  • Optogenetics / instrumentation*
  • Rats
  • Time Factors
  • Transfection

Substances

  • Channelrhodopsins
  • Glutamic Acid
  • Calcium

Grants and funding

PD received funding from the European Commission (http://ec.europa.eu/) under project OptoNeuro, project number 249867 (www.optoneuro.eu). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.