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Vol. 13, Issue 4, 1329-1337, April 2002

Regulated Disruption of Inositol 1,4,5-Trisphosphate Signaling in Caenorhabditis elegans Reveals New Functions in Feeding and Embryogenesis

Denise S. Walker,^* Nicholas J.D. Gower,^* Sung Ly,^* Gemma L. Bradley,^* and Howard A. Baylis^*§

 ^*Department of Zoology, University of Cambridge, Cambridge, CB2 3EJ, United Kingdom; and  Laboratory of Receptor Signaling, The Babraham Institute, Cambridge, CB2 4AT, United Kingdom

Inositol 1,4,5-trisphosphate (IP₃) is an important second messenger in animal cells and is central to a wide range of cellular responses. The major intracellular activity of IP₃ is to regulate release of Ca²⁺ from intracellular stores through IP₃ receptors (IP₃Rs). We describe a system for the transient disruption of IP₃ signaling in the model organism Caenorhabditis elegans. The IP₃ binding domain of the C. elegans IP₃R, ITR-1, was expressed from heat shock-induced promoters in live animals. This results in a dominant-negative effect caused by the overexpressed IP₃ binding domain acting as an IP₃ "sponge." Disruption of IP₃ signaling resulted in disrupted defecation, a phenotype predicted by previous genetic studies. This approach also identified two new IP₃-mediated processes. First, the up-regulation of pharyngeal pumping in response to food is dependent on IP₃ signaling. RNA-mediated interference studies and analysis of itr-1 mutants show that this process is also IP₃R dependent. Second, the tissue-specific expression of the dominant-negative construct enabled us to circumvent the sterility associated with loss of IP₃ signaling through the IP₃R and thus determine that IP₃-mediated signaling is required for multiple steps in embryogenesis, including cytokinesis and gastrulation.

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Present address: University College London, Department of Biochemistry and Medical Biology, Darwin Building, Gower St., London, WC1E 6BT, United Kingdom.

^§ Corresponding author. E-mail address: hab{at}mole.bio.cam.ac.uk.

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Molecular Biology of the Cell
Vol. 13, 1329-1337, April 2002
Copyright © 2002 by The American Society for Cell Biology

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