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Detection of sodium channel activators by a rapid fluorimetric microplate assay.
Chem Res Toxicol. 2004 Apr; 17(4):572-8.CR

Abstract

Marine toxins such as brevetoxins and ciguatoxins are produced by dinoflagellates and can accumulate in seafood. These toxins affect humans through seafood consumption. Intoxication is mainly characterized by gastrointestinal and neurological disorders and, in most severe cases, by cardiovascular problems. To prevent the consumption of food contaminated with these toxins, shellfish have been tested by mouse bioassay. However, this method is expensive, time-consuming, and ethically questionable. The objective of this study was to use a recently developed fluorimetric microplate assay to rapidly detect brevetoxins and ciguatoxins. The method is based on the pharmacological effect of brevetoxins and ciguatoxins known to activate sodium channels and involves (i). the incubation of excitable cells in 96 well microtiter plates with the fluorescent dye bis-oxonol, whose distribution across the membrane is potential-dependent, and (ii). dose-dependent cell depolarization by the toxins. Our findings demonstrate that measuring changes in membrane potential induced by brevetoxins and ciguatoxins allowed their quantitation. Active toxins could be reliably detected at concentrations in the nanomolar range. The simplicity, sensitivity, and possibility of being automated provide the basis for development of a practical alternative to conventional testing for brevetoxins and ciguatoxins.

Authors+Show Affiliations

Departamento de Farmacologia and Departamento de Fisiologia Animal, Facultad de Veterinaria de Lugo Universidad de Santiago de Compostela, 27002 Lugo, Spain.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

15089100

Citation

Louzao, M C., et al. "Detection of Sodium Channel Activators By a Rapid Fluorimetric Microplate Assay." Chemical Research in Toxicology, vol. 17, no. 4, 2004, pp. 572-8.
Louzao MC, Vieytes MR, Yasumoto T, et al. Detection of sodium channel activators by a rapid fluorimetric microplate assay. Chem Res Toxicol. 2004;17(4):572-8.
Louzao, M. C., Vieytes, M. R., Yasumoto, T., & Botana, L. M. (2004). Detection of sodium channel activators by a rapid fluorimetric microplate assay. Chemical Research in Toxicology, 17(4), 572-8.
Louzao MC, et al. Detection of Sodium Channel Activators By a Rapid Fluorimetric Microplate Assay. Chem Res Toxicol. 2004;17(4):572-8. PubMed PMID: 15089100.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Detection of sodium channel activators by a rapid fluorimetric microplate assay. AU - Louzao,M C, AU - Vieytes,M R, AU - Yasumoto,T, AU - Botana,L M, PY - 2004/4/20/pubmed PY - 2004/11/13/medline PY - 2004/4/20/entrez SP - 572 EP - 8 JF - Chemical research in toxicology JO - Chem Res Toxicol VL - 17 IS - 4 N2 - Marine toxins such as brevetoxins and ciguatoxins are produced by dinoflagellates and can accumulate in seafood. These toxins affect humans through seafood consumption. Intoxication is mainly characterized by gastrointestinal and neurological disorders and, in most severe cases, by cardiovascular problems. To prevent the consumption of food contaminated with these toxins, shellfish have been tested by mouse bioassay. However, this method is expensive, time-consuming, and ethically questionable. The objective of this study was to use a recently developed fluorimetric microplate assay to rapidly detect brevetoxins and ciguatoxins. The method is based on the pharmacological effect of brevetoxins and ciguatoxins known to activate sodium channels and involves (i). the incubation of excitable cells in 96 well microtiter plates with the fluorescent dye bis-oxonol, whose distribution across the membrane is potential-dependent, and (ii). dose-dependent cell depolarization by the toxins. Our findings demonstrate that measuring changes in membrane potential induced by brevetoxins and ciguatoxins allowed their quantitation. Active toxins could be reliably detected at concentrations in the nanomolar range. The simplicity, sensitivity, and possibility of being automated provide the basis for development of a practical alternative to conventional testing for brevetoxins and ciguatoxins. SN - 0893-228X UR - https://www.unboundmedicine.com/medline/citation/15089100/Detection_of_sodium_channel_activators_by_a_rapid_fluorimetric_microplate_assay_ L2 - https://doi.org/10.1021/tx0342262 DB - PRIME DP - Unbound Medicine ER -