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J. Biol. Chem., Vol. 282, Issue 19, 13984-13993, May 11, 2007

This Article Full Text Full Text (PDF) Supplemental Data All Versions of this Article: 282/19/13984    most recent M700940200v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Ashworth, R. Articles by Patel, S. Search for Related Content PubMed PubMed Citation Articles by Ashworth, R. Articles by Patel, S. Social Bookmarking                      What's this?

Molecular and Functional Characterization of Inositol Trisphosphate Receptors during Early Zebrafish Development^*

Rachel Ashworth¹, Benoit Devogelaere², Jez Fabes^¶, Richard E. Tunwell^¶, Kevin R. Koh^¶, Humbert De Smedt, and Sandip Patel^¶3

From the School of Biological and Chemical Sciences, Queen Mary University of London, London E1 4NS, United Kingdom, Laboratorium voor Fysiologie, Katholieke Universiteit Leuven, Leuven Campus Gasthuisberg O/N1, Herestraat 49, B-3000 Leuven, Belgium, and ^¶Department of Physiology, University College London, London WC1E 6BT, United Kingdom

Fluctuations in cytosolic Ca²⁺ are crucial for a variety of cellular processes including many aspects of development. Mobilization of intracellular Ca²⁺ stores via the production of inositol trisphosphate (IP₃) and the consequent activation of IP₃-sensitive Ca²⁺ channels is a ubiquitous means by which diverse stimuli mediate their cellular effects. Although IP₃ receptors have been well studied at fertilization, information regarding their possible involvement during subsequent development is scant. In the present study we examined the role of IP₃ receptors in early development of the zebrafish. We report the first molecular analysis of zebrafish IP₃ receptors which indicates that, like mammals, the zebrafish genome contains three distinct IP₃ receptor genes. mRNA for all isoforms was detectable at differing levels by the 64 cell stage, and IP₃-induced Ca²⁺ transients could be readily generated (by flash photolysis) in a controlled fashion throughout the cleavage period in vivo. Furthermore, we show that early blastula formation was disrupted by pharmacological blockade of IP₃ receptors or phospholipase C, by molecular inhibition of the former by injection of IRBIT (IP₃ receptor-binding protein released with IP₃) and by depletion of thapsigargin-sensitive Ca²⁺ stores after completion of the second cell cycle. Inhibition of Ca²⁺ entry or ryanodine receptors, however, had little effect. Our work defines the importance of IP₃ receptors during early development of a genetically and optically tractable model vertebrate organism.

Received for publication, January 31, 2007

^* This work was supported by a David Phillips Fellowship from the Biotechnology and Biological Sciences Research Council (to R. A.) and a Career Development Research Fellowship from the Wellcome Trust (to S. P.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains supplemental Figs. 1–4.

² A research assistant from the Fund for Scientific Research Flanders.

¹ To whom correspondence may be addressed: School of Biological and Chemical Sciences, Queen Mary University of London, Mile End Rd., London E1 4NS, United Kingdom. Tel.: 20-7882-3010; Fax: 20-8983-0973; E-mail: r.ashworth{at}qmul.ac.uk. ³ To whom correspondence may be addressed: Dept. of Physiology, University College London, Gower St., London, WC1E 6BT, United Kingdom. Tel.: 207-679-6540; Fax: 207-916-7968; E-mail: patel.s{at}ucl.ac.uk.

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