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The function of cyclic nucleotide-gateD ion channels in the cerebellum

Objectif



Until recently, cyclic nucleotide-gated ion channels (CNG channels) were thought to be of importance only in retinal photoreceptor cells and in olfactory cells, where they play key roles in sensory signal transduction. It is now clear that a family of such channels exists and that they are likely to be expressed in a wide variety of cell types. Little has been done, so far, to examine the possibility that CNG channels are utilised in the brain. Pilot investigations by members of the proposed partnership, however, point to a widespread distribution of these channels in the brain, with a notable concentration of them in Purkinje cells in the cerebellum. The cerebellum is one of the brain areas in which the diffusible messenger, nitric oxide (NO), is known to operate. NO induces cGMP accumulation in target cells through stimulation of soluble guanylyl cyclase and the Purkinje cell represents a likely target that responds in this way to NO generated in neighbouring structures.
The aim of the research is: (a) to identify the molecular composition of CNG channels in the cerebellum, to elucidate the functional properties of cloned CNG channel subunits when heterologously expressed in cell lines, and to compare these properties with those of native channels; (b) to identify the cerebellar cells that express mRNA for different channel subunits and to locate, at the cellular and subcellular levels, the channel proteins; (c) to generate novel chemical tools, including caged derivatives, that will be used for experimental investigation of CNG channels in single cells and (d) to test hypothesis relating to the role of CNG channels located postsynaptically (on Purkinje cell somata and dendrites) and presynaptically (on Purkinje cell axon terminals) in the regulation of synaptic transmission.
To accomplish these goals requires an integrated, multi-disciplinary effort. To this end, each member of the partnership will contribute particular expertise and the combined effort will provide the strength necessary to approach the project in a way that would otherwise not be feasible. Thus, U. Benjamin Kaupp (Julich, Germany), will identify known and novel channel subunits in the cerebellum and determine their functional properties, alone and in combination, using expression systems; he will also provide J. Bradley (Paris, France) and J. Garthwaite (London, UK) with probes and antibodies to determine the anatomical distribution of the channels; V. Hagen will design and synthesise caged cyclic nucleotide derivatives that will provide novel tools to probe the function of CNG channels; P. Ascher (Paris, France) and J. Garthwaite will investigate the function of CNG channels expressed pre- and postsynaptically in Purkinje cells.
Apart from the contribution that the work will make towards the basic understanding of this signalling pathway in brain function, the research may also shed new light on the roles of CNG channels in disease states, notably those (e.g. neurodegeneration associated with stroke) in which glutamate and NO have already been implicated.

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Coordinateur

Forschungszentrum Julich GmbH
Contribution de l’UE
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Adresse

52425 Jülich
Allemagne

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