Understanding the neuronal mechanisms mediating emotional information processing in the amygdala will require identification of its main cell types and knowledge of their participation in the intrinsic circuitry of this region. Amygdala pyramidal neurons a re spiny projection neurons that use glutamate as an excitatory neurotransmitter.
In contrast, most non-pyramidal neurons in the amygdala are aspiny interneurons that use GABA as an inhibitory neurotransmitter. Very few studies linking physiological and morphological properties of the various cell types of the amygdala have been done. The precise identification of the main cell types in the amygdala and knowledge of their participation in the intrinsic circuitry of this region is an essential pre-requisite to understand the neuronal mechanisms mediating the diverse roles of the amygdala subsystems.
The aim of this project is to characterise some electrophysiological and pharmacological properties of neurochemically and morphologically identified interneurones in the amygdala. This will be a multidisciplinary study in which intracellular recordings followed by intracellular filling will be combined with immunocytochemical characterisation and cellular reconstruction.
For this purpose, a transgenic mouse line expressing a green fluorecescent protein (GFP) under the control of the GAD65 (GAD: glutamate decarboxilase enzyme involved in the synthesis of GABA) promoter will be used. After the single, double or triple intracellular recordings, sections will be fixed and re-sectioned. Immunoreactivity for neurotransmitter receptors, calcium binding proteins and neuropeptides will be visualized by immunocytochemistry and the sections will be processed.
The axonal and dendritic patterns of each neurone will be then analysed at high magnification and reconstructed. Structural properties of interneuron to principal cell and principal cell to interneuron inputs will also be investigated by electron microscopic analysis.
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