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The Journal of Neuroscience, January 1, 2003, 23(1):64-72

Detection of Calcium Transients in Drosophila Mushroom Body Neurons with Camgaroo Reporters

Dinghui Yu¹, Geoffrey S. Baird³, Roger Y. Tsien^{3, 4}, and Ronald L. Davis^{1, 2}

Departments of ¹ Molecular and Cellular Biology and ² Psychiatry and Behavioral Sciences, Baylor College of Medicine, Houston, Texas 77030, and ³ Department of Pharmacology, ⁴ Howard Hughes Medical Institute, University of California, San Diego, San Diego, California 92093

Camgaroos are yellow fluorescent protein derivatives that hold promise as transgenically encoded calcium sensors in behaving animals. We expressed two versions of camgaroo in Drosophila mushroom bodies using the galactosidase-4 (GAL4) system. Potassium depolarization of brains expressing the reporters produces a robust increase in fluorescence that is blocked by removing extracellular calcium or by antagonists of voltage-dependent calcium channels. The fluorescence increase is not attributable to cytoplasmic alkalization; depolarization induces a slight acidification of the cytoplasm of mushroom body neurons. Acetylcholine applied near the dendrites of the mushroom body neurons induces a rapid and ipsilateral-specific fluorescence increase in the mushroom body axons that is blocked by antagonists of calcium channels or nicotinic acetylcholine receptors. Fluorescence was observed to increase in all three classes of mushroom body neurons, indicating that all types respond to cholinergic innervation.

Key words: mushroom bodies; calcium; optical imaging; Drosophila; camgaroos; GFP

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Copyright © 2003 Society for Neuroscience  0270-6474/03/23164-09$05.00/0

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