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Elizabeth R. Fairey,¹ J. Stewart G. Edmunds,¹ Nora J. Deamer-Melia,² Howard Glasgow Jr.,² Frank M. Johnson,³ Peter R. Moeller,¹ JoAnn M. Burkholder,² and John S. Ramsdell¹

¹Marine Biotoxins Program, Center for Coastal Environmental Health and Biomolecular Research, NOAA-National Ocean Service, Charleston, South Carolina, USA
²Department of Botany, North Carolina State University, Raleigh, North Carolina, USA
³Intramural Research Program and Environmental Toxicology Program, National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina, USA

Abstract

Collaborative studies were performed to develop a functional assay for fish-killing activity produced by Pfiesteria piscicida. Eight cell lines were used to screen organic fractions and residual water fraction by using a 3-[4,5-dimethylthiazol-(2-4) ]-diphenyltetrazolium bromide cytotoxicity assay. Diethyl ether and a residual water fraction were cytotoxic to several cell lines including rat pituitary (GH₄C₁) cells. Residual water as well as preextracted culture water containing P. piscicida cells induced c-fos-luciferase expressed in GH₄C₁ cells with a rapid time course of induction and sensitive detection. The reporter gene assay detected activity in toxic isolates of P. piscicida from several North Carolina estuaries in 1997 and 1998 and may also be suitable for detecting toxic activity in human and animal serum. Key words: assay, c-fos, GH₄C₁, Pfiesteria piscicida, pituitary, toxin. Environ Health Perspect 107:711-714 (1999) . [Online 28 July 1999]

http://ehpnet1.niehs.nih.gov/docs/1999/107p711-714fairey/ abstract.html

Address correspondence to J.S. Ramsdell, NOAA-NOS Center for Coastal Environmental Health and Biomolecular Research, 219 Fort Johnson Road, Charleston, SC 29412 USA. Telephone: (843) 762-8510. Fax: (843) 762-8700. E-mail: john.ramsdell@noaa.gov

We thank D. Xi for initial studies with the c-fos-luciferase reporter gene construct in GH₄C₁ cells and M. Snell for assistance during the two interlaboratory studies.

Received 8 December 1998 ; accepted 10 May 1999.