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1 Università di Ferrara, Dip. Biologia, Sezione di Fisiologia e Biofisica - Centro di Neuroscienze, Via Borsari, 46-44100 Ferrara, Italy
2 Fukushima Medical University School of Medicine, Department Molecular Genetics, Hikarigaoka, Fukushima 960-1295, Japan
3 Keio University, School of Medicine, Department of Physiology, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan
The olfactory bulb of mammals contains a large population of dopaminergic interneurones within the glomerular layer. Dopamine has been shown both in vivo and in vitro to modulate several aspects of olfactory information processing, but the functional properties of dopaminergic neurones have never been described due to the inability to recognize these cells in living preparations. To overcome this difficulty, we used a transgenic mouse strain harbouring an eGFP (enhanced green fluorescent protein) reporter construct under the promoter of tyrosine hydroxylase, the rate-limiting enzyme for cathecolamine synthesis. As a result, we were able to identify dopaminergic neurones (TH-GFP cells) in living preparations and, for the first time, we could study the functional properties of such neurones in the olfactory bulb, in both slices and dissociated cells. The most prominent feature of these cells was the autorhythmicity. In these cells we identified five main voltage-dependent conductances: the two having largest amplitude were a fast transient Na+ current and a delayed rectifier K+ current. In addition, we observed three smaller inward currents, sustained by Na+ ions (persistent type) and by Ca2+ ions (LVA and HVA). Using pharmacological tools and ion substitution methods we showed that the pacemaking process is supported by the interplay of the persistent Na+ current and of a T-type Ca2+ current. We carried out a complete kinetical analysis of the five conductances present in these cells, and developed a Hodgkin-Huxley model of TH-GFP cells, capable of reproducing accurately the properties of living cells, including autorhytmicity, and allowing a precise understanding of the process.
(Received 7 February 2005;
accepted after revision 22 February 2005;
first published online 24 February 2005)
Corresponding author O. Belluzzi: Dip. Biologia, Sezione di Fisiologia e Biofisica - Via Borsari, 46-44100 Ferrara, Italy. Email: mk5{at}unife.it
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