Reporter gene assay for fish-killing activity produced by Pfiesteria piscicida.

Fairey, Elizabeth R.; Edmunds, J. S. G.; Deamer-Melia, Nora J.; Glasgow, Howard B.; Johnson, Francis; Moeller, P. R.; Burkholder, JoAnn M.; Ramsdell, John S.

doi:10.1289/ehp.99107711

Cited by 66 publications

(64 citation statements)

References 15 publications

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“…Subsequent to elevating cytosolic free calcium, pPfTx induced the permeabilization of GH 4 C 1 cells to YO-PRO-1. This action of pPfTx is consistent with its cytotoxic effect (27) and the decreased c-fos luciferase activity (20). The permeabilizing effect of pPfTx in GH 4 C 1 cells was completely inhibited by oxATP.…”

Section: Discussionsupporting

confidence: 67%

“…Calcium is a second messenger, and one of its downstream signaling events is among those factors that induce the immediate response gene c-fos. For that reason, the sustained elevation of cytosolic free calcium by pPfTx is consistent with the induction of the c-fos (20,27). Activation of P2X 7 receptors leads to strong induction of additional transcription factors as well, including nuclear factor (NF)-kappaB and the nuclear factor of activated T cells (NFAT), both of which are associated with cytokine production (33).…”

Section: Discussionmentioning

confidence: 70%

“…A bioactive substance has been identified in toxic Pfiesteria cultures with cytotoxic selectivity to GH 4 C 1 cells based upon a panel of different cell types (27). We report here that the pPfTx elevates cytosolic free calcium in Fura2-loaded GH 4 C 1 cells.…”

Section: Discussionmentioning

confidence: 71%

“…The putative Pfiesteria toxin (pPfTx) also elevated cytosolic free calcium in Fura-2 in GH 4 C 1 cells and increased the permeability to YO-PRO-1 in a manner inhibited fully by oxATP. This study indicates that GH 4 concentrations induces a c-fos luciferase reporter gene (27). The expression of c-fos in neural cells is often associated with elevated cytosolic free calcium [Ca 2+ ] i (28).…”

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confidence: 70%

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Microfluorimetric analysis of a purinergic receptor (P2X7) in GH4C1 rat pituitary cells: effects of a bioactive substance produced by Pfiesteria piscicida.

Melo

Moeller

Glasgow

et al. 2001

Environ Health Perspect

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Pfiesteria piscicida Steidinger & Burkholder is a toxic dinoflagellate that leads to fish and human toxicity. It produces a bioactive substance that leads to cytotoxicity of GH4C1 rat pituitary cells. Extracellular adenosine 5'-triphosphate (ATP) acting on P2X7 purinergic receptors induces the formation of a nonselective cation channel, causing elevation of the cytosolic free calcium followed by a characteristic permeabilization of the cell to progressively larger ions and subsequent cell lysis. We investigated whether GH4C1 rat pituitary cells express functional P2X7 receptors, and if so, are they activated by a bioactive substance isolated from toxic P. piscicida cultures. We tested the selective agonist 2'-3'-O-(benzoyl-4-benzoyl)-ATP (BzATP) and antagonists piridoxalphosphate-6-azophenyl-2'-4'-disulfonic acid (PPADS) and oxidized-ATP (oxATP) using elevated cytosolic free calcium in Fura-2 loaded cells, and induced permeability of these cells to the fluorescent dye YO-PRO-1 as end points. We demonstrated that in GH4C1 cells, BzATP induces both the elevation of cytosolic free calcium and the permeabilization of the cell membrane. ATP-induced membrane permeabilization was inhibited by PPADS reversibly and by oxATP irreversibly. The putative Pfiesteria toxin (pPfTx) also elevated cytosolic free calcium in Fura-2 in GH4C1 cells and increased the permeability to YO-PRO-1 in a manner inhibited fully by oxATP. This study indicates that GH4C1 cells express a purinoceptor with characteristics consistent with the P2X7 subtype, and that pPfTx mimics the kinetics of cell permeabilization by ATP.

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Section: Discussionsupporting

confidence: 67%

Section: Discussionmentioning

confidence: 70%

Section: Discussionmentioning

confidence: 71%

mentioning

confidence: 70%

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Microfluorimetric analysis of a purinergic receptor (P2X7) in GH4C1 rat pituitary cells: effects of a bioactive substance produced by Pfiesteria piscicida.

Melo

Moeller

Glasgow

et al. 2001

Environ Health Perspect

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“…Although the acquisition of PEAS was temporally associated with exposure to estuarine waters inhabited by TPC, no data directly establish a causal relationship between exposure to TPC toxin(s) and development of PEAS. The definitive establishment of PEAS as a TPC toxin-mediated illness must await delineation of the toxin(s) structure and a rigorous test for its presence in biologic tissue (17). Previous cases thought to be due to environmental TPC toxin(s) exposure occurred in warm weather near the time of large fish kills (14).…”

Section: Discussionmentioning

confidence: 99%

Possible estuary-associated syndrome: symptoms, vision, and treatment.

Shoemaker

Hudnell

2001

Environ Health Perspect

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The human illness designated as possible estuarine-associated syndrome (PEAS) by the Centers for Disease Control and Prevention (CDC) has been associated with exposure to estuaries inhabited by toxin-forming dinoflagellates, including members of the fish-killing toxic Pfiesteria complex (TPC), Pfiesteria piscicida and Pfiesteria shumwayae. Humans may be exposed through direct contact with estuarine water or by inhalation of aerosolized or volatilized toxin(s). The five cases reported here demonstrate the full spectrum of symptoms experienced during acute and chronic stages of this suspected neurotoxin-mediated illness. The nonspecific symptoms most commonly reported are cough, secretory diarrhea, headache, fatigue, memory impairment, rash, difficulty in concentrating, light sensitivity, burning skin upon water contact, muscle ache, and abdominal pain. Less frequently encountered symptoms are upper airway obstruction, shortness of breath, confusion, red or tearing eyes, weakness, and vertigo. Some patients experience as few as four of these symptoms. The discovery that an indicator of visual pattern-detection ability, visual contrast sensitivity (VCS), is sharply reduced in affected individuals has provided an objective indicator that is useful in diagnosing and monitoring PEAS. VCS deficits are present in both acute and chronic PEAS, and VCS recovers during cholestyramine treatment coincident with symptom abatement. Although PEAS cannot yet be definitively associated with TPC exposure, resolution with cholestyramine treatment suggests a neurotoxin-mediated illness.

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Pfiesteria: The Toxic Pfiesteria Complex

Burkholder¹

2003

Encyclopedia of Environmental Microbiology

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General Characteristics, Systematics, and Life Cycle Systematics and Cellular Characteristics Complex Life Cycles Culturing Pfiesteria Assessment of Toxic Pfiesteria Involvement in Fish Kills Distribution, Nutritional Ecology, and Predation Sublethal and Chronic Impacts of Toxic Pfiesteria on Fish and Mammalian Health

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Reporter gene assay for fish-killing activity produced by Pfiesteria piscicida.

Cited by 66 publications

References 15 publications

Microfluorimetric analysis of a purinergic receptor (P2X7) in GH4C1 rat pituitary cells: effects of a bioactive substance produced by Pfiesteria piscicida.

Microfluorimetric analysis of a purinergic receptor (P2X7) in GH4C1 rat pituitary cells: effects of a bioactive substance produced by Pfiesteria piscicida.

Possible estuary-associated syndrome: symptoms, vision, and treatment.

Pfiesteria: The Toxic Pfiesteria Complex

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