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Plant and Cell Physiology Advance Access published online on April 13, 2005
Plant and Cell Physiology, doi:10.1093/pcp/pci102
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Plant and Cell Physiology 2005 © The Japanese Society of Plant Physiologists (JSPP); all rights reserved.
Received November 24, 2004
Accepted April 1, 2005

Short Communication

Monitoring Cytosolic Calcium in the Dinoflagellate Crypthecodinium Cohnii with Calcium Orange-AM

Connie M. C. Lam. Patrick 1, K. K. Yeung 1, and Joseph T. Y. Wong 1*

1 Biology Department, Hong Kong University of Science and Technology, Clearwater Bay, Kowloon, Hong Kong SAR, People's Republic of China

* To whom correspondence should be addressed.
Joseph T. Y. Wong, E-mail: botin{at}Ust.hk


  Abstract

Calcium plays several important roles in the signal transduction pathways of dinoflagellates. We describe here the development of calcium orange-AM as an intracellular calcium reporter for the heterotrophic dinoflagellate Crypthecodinium cohnii. We demonstrated with confocal microscopy that by restricting the incubation period to 30 to 45 minutes, no compartmentalization of the dye occurs in the mitochondria or endoplasmic reticulum. The dye fluorescence responded well to the effects of calcium ionophores and calcium chelators. By calibrating the dye with known calcium concentrations, we determined the intracellular calcium concentration of C.cohnii to be 158 ± 56 nM, which rose to about 550 nM upon mechanical stimulation. [103]

Keywords: dinoflagellates; intracellular calcium; mechanical stimulation.
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