Hostname: page-component-78c5997874-j824f Total loading time: 0 Render date: 2024-11-16T09:20:14.476Z Has data issue: false hasContentIssue false
  • English
  • Français

Multiplex fluorimetric assays for monitoring algal toxins

Published online by Cambridge University Press:  02 December 2008

Carmen K.M. Mak ,
Patrick K.K. Yeung ,
Alvin C.M. Kwok ,
Y.H. Wong  and
Joseph T.Y. Wong
Carmen K.M. Mak
Affiliation:
Biology Department, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong
Patrick K.K. Yeung
Affiliation:
Biology Department, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong
Alvin C.M. Kwok
Affiliation:
Biology Department, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong
Y.H. Wong
Affiliation:
Biotechnology Research Institute and Biochemistry Department, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong
Joseph T.Y. Wong*
Affiliation:
Biology Department, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong
*
Correspondence should be addressed to: J.T.Y. Wong, Biology Department, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong email: [email protected]
Get access Rights & Permissions [Opens in a new window]

Abstract

Most known algal toxins act on ion channels either directly or indirectly, resulting in a change in intracellular ion concentrations when administered to targeted cells. The present project developed the working conditions for the use of fluorescent dyes in monitoring changes in membrane potential, intracellular calcium, and intracellular sodium levels in mammalian cell lines. Using these conditions, we were able to demonstrate specific changes in fluorescent signals in response to several purified toxins. We were also able to generate algal extracts which, when administered to the developed fluorimetric assays, were able to elicit different pattern of changes in membrane potential, intracellular calcium, and intracellular sodium levels. The differential pattern of responses induced by the different algal toxins in the three fluorimetric assays serve as a proof of concept for the use of multiplex fluorimetric assays in the laboratory monitoring of algal toxins.

Keywords

multiplex fluorimetric assaysmonitoringalgal toxins
Type
Research Article
Information
Journal of the Marine Biological Association of the United Kingdom , Volume 89 , Issue 2 , March 2009 , pp. 287 - 293
Copyright
Copyright © Marine Biological Association of the United Kingdom 2008

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Baden, D.G. and Adams, D.J. (2000) Brevetoxins: chemistry, mechanism of action, and methods of detection. New York: Marcel Dekker, Inc.CrossRefGoogle Scholar
Berman, F.W. and Murray, T.F. (1999) Brevetoxins cause acute excitotoxicity in primary cultures of rat cerebellar granule neurons. Journal of Pharmacology and Experimental Therapeutics 290, 439–444.Google ScholarPubMed
Bicalho, A.F., Guatimosim, C., Prado, M.A., Gomez, M.V. and Romano-Silva, M.A. (2002) Investigation of the modulation of glutamate release by sodium channels using neurotoxins. Neuroscience 113, 115–123.CrossRefGoogle ScholarPubMed
Bourdelais, A.J., Campbell, S., Jacocks, H., Naar, J., Wright, J.L., Carsi, J. and Baden, D.G. (2004) Brevenal is a natural inhibitor of brevetoxin action in sodium channel receptor binding assays. Cellular and Molecular Neurobiology 24, 553–563.Google Scholar
Bourdelais, A.J., Campbell, S., Jacocks, H., Naar, J., Wright, J.L., Carsi, J. and Campora, C.E., Hokama, Y. and Ebesu, J.S. (2006) Comparative analysis of purified Pacific and Caribbean ciguatoxin congeners and related marine toxins using a modified ELISA technique. Journal of Clinical Laboratory Analysis 20, 121–125.Google Scholar
Cestele, S. and Catterall, W.A. (2000) Molecular mechanisms of neurotoxin action on voltage-gated sodium channels. Biochimie 82, 883–892.Google Scholar
Cestele, S., Sampieri, F., Rochat, H. and Gordon, D. (1996) Tetrodotoxin reverses brevetoxin allosteric inhibition of scorpion α-toxin binding on rat brain sodium channels. Journal of Biological Chemistry 271, 18329–18332.Google Scholar
Dechraoui, M.B., Naar, J., Pauillac, S. and Legrand, A.M. (1999) Ciguatoxins and brevetoxins, neurotoxic polyether compounds active on sodium channels. Toxicon 37, 125–143.Google Scholar
Dechraoui, M.Y., Wacksman, J.J. and Ramsdell, J.S. (2006) Species selective resistance of cardiac muscle voltage gated sodium channels: characterization of brevetoxin and ciguatoxin binding sites in rats and fish. Toxicon 48, 702–712.Google Scholar
Diarra, A., Sheldon, C. and Church, J. (2001) In situ calibration and sensitivity of [H+]the fluorescent Na+ indicator SBFI. American Journal of Physiology—Cell Physiology 280, C1623–1633.CrossRefGoogle ScholarPubMed
Dickey, R., Jester, E., Granade, R., Mowdy, D., Moncreiff, C., Rebarchik, D., Robl, M., Musser, S. and Poli, M. (1999) Monitoring brevetoxins during a Gymnodinium Breve red tide: comparison of osdium channel specificity cytotoxicity assay and mouse bioassay for determination of neurotoxic shellfish toxins in shellfish extracts. Natural Toxins 7, 157–165.Google Scholar
Escobar, L.I., Salvador, C., Martinez, M. and Vaca, L. (1998) Maitotoxin, a cationic channel activator. Neurobiology 6, 59–74.Google Scholar
Fairey, E.R., Dechraoui, M.B., Sheets, M.F. and Ramsdell, J.S. (2001) Modification of the cell based assay for brevetoxins using human cardiac voltage dependent sodium channels expressed in HEK-293 cells. Biosensors and Bioelectronics 16, 579–586.Google Scholar
FAO (2004) Ciguatera fish poisoning. Rome: Food and Agriculture Organization of the United Nations.Google Scholar
Fernandez, M.L. and Cembella, A.D. (1995) Manual on harmful marine microalgae. Paris: UNESCO.Google Scholar
Gusovsky, F., Yasumoto, T. and Daly, J.W. (1989) Maitotoxin, a potent, general activator of phosphoinositide breakdown. FEBS Letters 243, 307–312.Google Scholar
Guzman-Perez, S.E. and Park, D.L. (2000) Ciguatera toxins: chemistry and detection. New York: Marcel Dekker, Inc.Google Scholar
Inoue, M., Hirama, M., Satake, M., Sugiyama, K. and Yasumoto, T. (2003) Inhibition of brevetoxin binding to the voltage-gated sodium channel by gambierol and gambieric acid-A. Toxicon 41, 469–474.CrossRefGoogle Scholar
Jellett, J.F., Marks, L.J.E., Stewart, J., Dorey, M.L., Watson-Wright, W. and Lawrence, J.F. (1992) Paralytic shellfish poison (saxitoxin family) bioassays: automated endpoint determination and standardization of the in vitro tissue culture bioassay, and comparison with the standard mouse bioassay. Toxicon 30, 1143–1156.CrossRefGoogle ScholarPubMed
Lehane, L. and Lewis, R.J. (2000) Ciguatera: recent advances but the risk remains. International Journal of Food Microbiology 61, 91–125.Google Scholar
Louzao, M.C., Cagide, E., Vieytes, M.R., Sasaki, M., Fuwa, H., Yasumoto, T. and Botana, L.M. (2006) The sodium channel of human excitable cells is a target for gambierol. Cellular Physiology and Biochemistry 17, 257–268.Google Scholar
Maaser, K., Hopfner, M., Kap, H., Sutter, A.P., Barthel, B., von Lampe, B., Zeitz, M. and Scherubl, H. (2002) Extracellular nucleotides inhibit growth of human oesophageal cancer cells via P2Y(2)-receptors. British Journal of Cancer 86, 636–644.CrossRefGoogle ScholarPubMed
Manger, R.L., Leja, L.S., Lee, S.Y., Hungerford, J.M. and Wekell, M.M. (1993) Tetrazolium-based cell bioassay for neurotoxins active on voltage-sensitive sodium channels: semiautomated assay for saxitoxins, brevetoxins, and ciguatoxins. Analytical Biochemistry 214, 190–194.Google Scholar
Manger, R.L., Lee, S.Y., Leja, L.S., Hungerford, J.M., Hokama, Y., Dickey, R.W., Granade, H.R., Lewis, R., Yasumoto, T. and Wekell, M.M. (1995) In vitro analysis of ciguatoxins, brevetoxins, and saxitoxins. Koala, HI: Asian Pacific Research Foundation.Google Scholar
Merritt, J.E., McCarth, S.A., Davies, M.P. and Moores, K.E. (1990) Use of fluo-3 to measure cytosolic Ca2+ in platelets and neutrophils. Loading cells with the dye, calibration of traces, measurements in the presence of plasma, and buffering of cytosolic Ca2+. Biochemical Journal 269, 513–519.Google Scholar
Morales-Tlalpan, V. and Vaca, L. (2002) Modulation of the maitotoxin response by intracellular and extracellular cations. Toxicon 40, 493–500.Google Scholar
Murata, M., Naoki, H., Matsunaga, S., Satake, M. and Yasumoto, T. (1994) Structure and partial stereochemical assignments for maitotoxin, the most toxic and largest natural non-biopolymer. Journal of the American Chemical Society 116, 7098–7107.CrossRefGoogle Scholar
Trevino, C.L., De la Vega-Beltran, J.L., Nishigaki, T., Felix, R. and Darszon, A. (2006) Maitotoxin potently promotes Ca2+ influx in mouse spermatogenic cells and sperm, and induces the acrosome reaction. Journal of Cellular Physiology 206, 449–456.CrossRefGoogle ScholarPubMed
Van Dolah, F.M., Finley, E.L., Haynes, B.L., Doucette, G.J., Moeller, P.D. and Ramsdell, J.S. (1994) Development of rapid and sensitive high throughput pharmacologic assays for marine phycotoxins. Natural Toxins 2, 189–196.Google Scholar
Viviani, R. (1992) Eutrophication, marine biotoxins, human health. Science of the Total Environment (Supplement), 631–662.Google Scholar
Wang, S.Y. and Wang, G.K. (2003) Voltage-gated sodium channels as primary targets of diverse lipid-soluble neurotoxins. Cellular Signalling 15, 151–159.Google Scholar