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Spatiotemporal calcium signaling in a Drosophila melanogaster cell line stably expressing a Drosophila muscarinic acetylcholine receptor

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Invertebrate Neuroscience

Abstract

A muscarinic acetylcholine receptor (mAChR), DM1, expressed in the nervous system of Drosophila melanogaster, has been stably expressed in a Drosophila S2 cell line (S2-DM1) and used to investigate spatiotemporal calcium changes following agonist activation. Carbamylcholine (CCh) and oxotremorine are potent agonists, whereas application of the vertebrate M1 mAChR agonist, McN-A-343, results in a weak response. Activation of S2-DM1 receptors using CCh resulted in an increase in intracellular calcium ([Ca2+]i) that was biphasic. Two distinct calcium sources were found to contribute to calcium signaling: (1) internal stores that are sensitive to both thapsigargin and 2-aminoethoxydiphenyl borate and (2) capacitative calcium entry. Spatiotemporal imaging of individual S2-DM1 cells showed that the CCh-induced [Ca2+]i transient resulted from a homogeneous calcium increase throughout the cell, indicative of calcium release from internal stores. In contrast, ionomycin induced the formation of a "calcium ring" at the cell periphery, consistent with external calcium influx.

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Acknowledgements

The authors acknowledge the financial support of the Medical Research Council of the United Kingdom.

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  1. V. Raymond Delpech

    Present address: Centre de Recherches sur la Cognition Animale, Université Paul Sabatier, 118 Route de Narbonne, 31062, Toulouse Cedex, France

Authors and Affiliations

  1. Dupont Crop Protection Products, Stine Haskell Research Center, Elkton Road, Newark, Delaware, 19714, USA

    D. Cordova & J. J. Rauh

  2. MRC Functional Genetics Unit, Department of Human Anatomy and Genetics, University of Oxford, South Parks Road, Oxford, OX1 3QX, UK

    V. Raymond Delpech & D. B. Sattelle

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  1. D. Cordova
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  2. V. Raymond Delpech
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Correspondence to D. Cordova.

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Cordova, D., Delpech, V.R., Sattelle, D.B. et al. Spatiotemporal calcium signaling in a Drosophila melanogaster cell line stably expressing a Drosophila muscarinic acetylcholine receptor. Invert Neurosci 5, 19–28 (2003). https://doi.org/10.1007/s10158-003-0024-2

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