Objective
The understanding of the functional consequences of neuronal diversity requires a precise identification of neuronal types to study the modalities of their synaptic communications and of their specific contributions to the function of the brain. To investigate these issues, we propose to express specifically fluorescent and luminescent reporter transgenes in a restricted neuronal subtype in order to identify and monitor these cells selectively among the network.
Interneurons, which regulate locally the function of neuronal networks by synchronising the activities of individual neurons, are characterised by an extreme diversity at the biochemical, functional and morphological levels. The current investigations of neocortical interneurons by Rossier using single cell RT-PCR after patch-clamp in acute slices, show that the expression of the neuropeptide somatostatin (SS) defines a population of interneurons which have common molecular and functional properties but cannot be recognized by their morphology in-vivo. The fact that this population constitutes a major inhibitory level acting on neocortical efferent pyramidal neurons (layer V) is also of great interest in this project.
The aim of the proposal is to divert the strong SS promoter to drive the specific expression of transgene reporters in these interneurons. The reporters are Green Fluorescent Protein (GFP, fluorescent), and Aequorin (luminescent upon increase of intracellular calcium)/ which both have been used extensively by Pozzan. A transgenic rodent line expressing GFP has recently been reported. The SS containing interneurons expressing GFP will be identified in acute slices by their fluorescence. This will greatly facilitate their study in the context of the local network by electrophysiology and molecular techniques such as single cell RT-PCR. If they alternatively express Aequorin, the intracellular calcium changes induced by their activity will result in light emission. The activity of this population, and thus of this specific level of the network will be investigated by monitoring the emitted light. Transgenes have previously been expressed within specific regions of the brain. However, the use of reporters detectable in living cells, enabling to monitor the activity of these cells represents a major technical advance in the study of the functional diversity of the CNS cellular network. This has widespread biological relevance as a basic research tool and ultimately for the development of pharmaceutical agents with greater pharmacological specificity.
Fields of science (EuroSciVoc)
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
- natural sciences chemical sciences inorganic chemistry alkaline earth metals
- natural sciences biological sciences biochemistry biomolecules proteins
- engineering and technology electrical engineering, electronic engineering, information engineering information engineering telecommunications telecommunications networks mobile network
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Coordinator
75231 PARIS
France
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