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The Journal of Neuroscience, July 15, 2002, 22(14):5856-5864

Synaptic Calcium-Channel Function in Drosophila: Analysis and Transformation Rescue of Temperature-Sensitive Paralytic and Lethal Mutations of Cacophony

Fumiko Kawasaki, Stephen C. Collins, and Richard W. Ordway

Department of Biology, The Pennsylvania State University, University Park, Pennsylvania 16802

Voltage-gated calcium channels play a key role in chemical synaptic transmission by providing the calcium trigger for regulated neurotransmitter release. Genes encoding the primary structural subunit, 1, as well as accessory subunits of presynaptic calcium channels have now been identified in a variety of organisms. The cacophony (cac) gene in Drosophila, also known as nightblind A, encodes a voltage-gated calcium-channel 1 subunit homologous to vertebrate 1 subunits implicated in neurotransmitter release. A recent genetic screen in our laboratory isolated cac^TS2, a conditional cac mutant exhibiting rapid paralysis at elevated temperatures. This mutant has allowed synaptic electrophysiology after acute perturbation of a specific calcium-channel gene product, demonstrating that cac encodes a primary calcium channel functioning in neurotransmitter release. Here we report the molecular lesion in cac^TS2, a missense mutation within a calcium-dependent regulatory domain of the 1 subunit, as well as phenotypic rescue of temperature-sensitive and lethal cac mutations by transgenic expression of a wild-type cac cDNA. Notably, rescue of rapid, calcium-triggered neurotransmitter release was achieved by neural expression of a single cDNA containing a subset of alternative exons and lacking any conserved synaptic-protein interaction sequence. Possible implications of these findings are discussed in the context of structure-function studies of synaptic calcium channels, as well as alternative splicing and mRNA editing of the cac transcript.

Key words: synapse; calcium channel; Drosophila; temperature-sensitive; alternative splicing; SYNPRINT

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Copyright © 2002 Society for Neuroscience  0270-6474/02/22145856-09$05.00/0

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