Identification of a P2X7 receptor in GH4C1 rat pituitary cells:: A potential target for a bioactive substance produced by Pfiesteria piscicida

被引:33
作者
Kimm-Brinson, KL
Moeller, PDR
Barbier, M
Glasgow, H
Burkholder, JM
Ramsdell, JS
机构
[1] NOAA, Coastal Res Branch,Ctr Coastal Environm Hlth & Bi, Natl Ocean Serv, Marine Biotoxins Program, Charleston, SC 29412 USA
[2] N Carolina State Univ, Dept Bot, Raleigh, NC 27695 USA
关键词
c-fos; GH(4)C(1); P2X7; Pfiesteria; pituitary; purinergic; toxin;
D O I
10.2307/3454703
中图分类号
X [环境科学、安全科学];
学科分类号
08 ; 0830 ;
摘要
We examined the pharmacologic activity of a putative toxin (pPfTx) produced by Pfesteria piscicida by characterizing the signaling pathways that induce the c-fos luciferase construct in GH(4)C(1) rat pituitary cells. Adenosine-5 ' -triphosphate (ATP) was determined to increase and, at higher concentrations, decrease luciferase activity in GH(4)C(1) rat pituitary cells that stably express c-fos luciferase. The inhibition of luciferase results from cytotoxicity, characteristic of the putative P. piscicida toxin (pPfTx). The actions of both pPfTx and ATP to induce c-fos luciferase were inhibited by the purinogenic receptor antagonist pyridoxalphosphate-6-azophenyl-2 ' ,4 ' -disulfonic acid (PPADS). Further characterization of a P2X receptor on the GH(4)C(1) cell was determined by the analog selectivity of P2X agonists. The P2X1/P2X3 agonist alpha,beta -methylene ATP (alpha,beta -MeATP) failed to increase or decrease c-fos luciferase. However, the P2X7 agonist 2 ' ,3 '-(4-benzoyl)benzoyl ATP (BzATP), which had a predominant cytotoxic effect, was more potent than ATP. Immunoblot analysis of GH(4)C(1) cell membranes confirmed the presence of a 70-kDa protein that was immunoreactive to an antibody directed against the carboxy-terminal domain unique to the P2X7 receptor. The P2X7 irreversible antagonist oxidized-ATP (oxATP) inhibited the action of ATP, BzATP, and pPfTx. These findings indicate that GH(4)C(1) cells express purinogenic receptors with selectivity consistent with the P2X7 subtype and that this receptor pathway mediates the induction of the c-fos luciferase reporter gene by ATP and the putative Pfiesteria toxin.
引用
收藏
页码:457 / 462
页数:6
相关论文
共 39 条
[1]  
Barnett George A., 1996, J WORLD SYSTEMS RES, V2, P1
[2]   Trophic controls on stage transformations of a toxic ambush-predator dinoflagellate [J].
Burkholder, JM ;
Glasgow, HB .
JOURNAL OF EUKARYOTIC MICROBIOLOGY, 1997, 44 (03) :200-205
[3]   NEW PHANTOM DINOFLAGELLATE IS THE CAUSATIVE AGENT OF MAJOR ESTUARINE FISH KILLS [J].
BURKHOLDER, JM ;
NOGA, EJ ;
HOBBS, CH ;
GLASGOW, HB .
NATURE, 1992, 358 (6385) :407-410
[4]   Pfiesteria piscicida and other Pfiesteria-like dinoflagellates: Behavior, impacts, and environmental controls [J].
Burkholder, JM ;
Glasgow, HB .
LIMNOLOGY AND OCEANOGRAPHY, 1997, 42 (05) :1052-1075
[5]  
Burkholder JM, 1998, ECOL APPL, V8, pS37
[6]   ATP INDUCES NUCLEOTIDE PERMEABILITY IN RAT MAST-CELLS [J].
COCKCROFT, S ;
GOMPERTS, BD .
NATURE, 1979, 279 (5713) :541-542
[7]   Tissue distribution of the P2X(7) receptor [J].
Collo, G ;
Neidhart, S ;
Kawashima, E ;
KoscoVilbois, M ;
North, RA ;
Buell, G .
NEUROPHARMACOLOGY, 1997, 36 (09) :1277-1283
[8]  
Di Virgilio F, 1999, PROG BRAIN RES, V120, P355
[9]   THE P2Z PURINOCEPTOR - AN INTRIGUING ROLE IN IMMUNITY, INFLAMMATION AND CELL-DEATH [J].
DIVIRGILIO, F .
IMMUNOLOGY TODAY, 1995, 16 (11) :524-528
[10]  
Fairey ER, 1999, NAT TOXINS, V7, P415, DOI 10.1002/1522-7189(199911/12)7:6<415::AID-NT81>3.0.CO