PUBLICATION
"In vivo" monitoring of neuronal network activity in zebrafish by two-photon Ca(2+) imaging
- Authors
- Brustein, E., Marandi, N., Kovalchuk, Y., Drapeau, P., and Konnerth, A.
- ID
- ZDB-PUB-030730-10
- 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.
Citation
Brustein, E., Marandi, N., Kovalchuk, Y., Drapeau, P., and Konnerth, A. (2003) "In vivo" monitoring of neuronal network activity in zebrafish by two-photon Ca(2+) imaging. Pflugers Archiv : European journal of physiology. 446(6):766-773.
Abstract
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping