Community Action Needed: Please respond to the NIH RFI
ZFIN ID: ZDB-PUB-030730-10
"In vivo" monitoring of neuronal network activity in zebrafish by two-photon Ca(2+) imaging
Brustein, E., Marandi, N., Kovalchuk, Y., Drapeau, P., and Konnerth, A.
Date: 2003
Source: Pflugers Archiv : European journal of physiology   446(6): 766-773 (Journal)
Registered Authors: Brustein, Edna, Drapeau, Pierre
Keywords: none
MeSH Terms:
  • Animals
  • Calcium/chemistry
  • Calcium/physiology*
  • Calcium Signaling/drug effects
  • Calcium Signaling/physiology
  • Coloring Agents
  • Diagnostic Imaging*
  • Excitatory Amino Acids/antagonists & inhibitors
  • Excitatory Amino Acids/pharmacology
  • Fluorescent Dyes
  • Glycine Agents/pharmacology
  • In Vitro Techniques
  • Larva/physiology
  • Nerve Net/drug effects
  • Nerve Net/growth & development
  • Nerve Net/physiology*
  • Neurons/physiology
  • Spinal Cord/cytology
  • Spinal Cord/growth & development
  • Spinal Cord/physiology
  • Strychnine/pharmacology
  • Zebrafish/physiology*
PubMed: 12883893 Full text @ Pflügers Archiv. / Eur. J. Physiol.
The zebrafish larva is a powerful model for the analysis of behaviour and the underlying neuronal network activity during early stages of development. Here we employ a new approach of "in vivo" Ca(2+) imaging in this preparation. We demonstrate that bolus injection of membrane-permeable Ca(2+) indicator dyes into the spinal cord of zebrafish larvae results in rapid staining of essentially the entire spinal cord. Using two-photon imaging, we could monitor Ca(2+) signals simultaneously from a large population of spinal neurons with single-cell resolution. To test the method, Ca(2+) transients were produced by iontophoretic application of glutamate and, as observed for the first time in a living preparation, of GABA or glycine. Glycine-evoked Ca(2+) transients were blocked by the application of strychnine. Sensory stimuli that trigger escape reflexes in mobile zebrafish evoked Ca(2+) transients in distinct neurons of the spinal network. Moreover, long-term recordings revealed spontaneous Ca(2+) transients in individual spinal neurons. Frequently, this activity occurred synchronously among many neurons in the network. In conclusion, the new approach permits a reliable analysis with single-cell resolution of the functional organisation of developing neuronal networks.