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  • Flash photolysis of caged compounds in the cilia of olfactory sensory neurons.

Flash photolysis of caged compounds in the cilia of olfactory sensory neurons.

Journal of visualized experiments : JoVE (2011-11-09)
Anna Boccaccio, Claudia Sagheddu, Anna Menini
ABSTRACT

Photolysis of caged compounds allows the production of rapid and localized increases in the concentration of various physiologically active compounds. Caged compounds are molecules made physiologically inactive by a chemical cage that can be broken by a flash of ultraviolet light. Here, we show how to obtain patch-clamp recordings combined with photolysis of caged compounds for the study of olfactory transduction in dissociated mouse olfactory sensory neurons. The process of olfactory transduction (Figure 1) takes place in the cilia of olfactory sensory neurons, where odorant binding to receptors leads to the increase of cAMP that opens cyclic nucleotide-gated (CNG) channels. Ca entry through CNG channels activates Ca-activated Cl channels. We show how to dissociate neurons from the mouse olfactory epithelium and how to activate CNG channels or Ca-activated Cl channels by photolysis of caged cAMP or caged Ca. We use a flash lamp to apply ultraviolet flashes to the ciliary region to uncage cAMP or Ca while patch-clamp recordings are taken to measure the current in the whole-cell voltage-clamp configuration.

MATERIALS
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Supelco
用于 ISE 的钙离子溶液, 0.1 M Ca, analytical standard (for ion-selective electrodes)