Optical probing of neuronal circuits with calcium indicators

Proc Natl Acad Sci U S A. 2000 Mar 28;97(7):3619-24. doi: 10.1073/pnas.97.7.3619.

Abstract

An experimental difficulty in unraveling circuits in the mammalian nervous system is the identification of postsynaptic targets of a given neuron. Besides ultrastructural reconstructions, simultaneous recordings from pairs of cells in brain slices have been used to identify connected neurons. We describe in this paper a technique using calcium imaging that allows rapid identification of potential postsynaptic targets. This method consists of stimulating one neuron ("trigger") while imaging a population of cells to detect which other neurons ("followers") are activated by the trigger. By using bulk-loading of calcium indicators in slices of mouse visual cortex, we demonstrate that neurons that display somatic calcium transients time-locked to the spikes of a trigger neuron can be monosynaptically connected to it. This technique could be applied to reconstruct and assay circuits in the central nervous system.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Calcium / physiology*
  • Fluorescent Dyes
  • Fura-2 / analogs & derivatives
  • In Vitro Techniques
  • Magnesium
  • Mice
  • Mice, Inbred C57BL
  • Neurons / physiology*
  • Optics and Photonics
  • Visual Cortex / cytology
  • Visual Cortex / physiology*

Substances

  • Fluorescent Dyes
  • fura-2-am
  • Magnesium
  • Calcium
  • Fura-2