Identification and characterization of an endogenous P2X7 (P2Z) receptor in CHO-K1 cells

被引:39
作者
Michel, AD [1 ]
Chessell, IP [1 ]
Hibell, AD [1 ]
Simon, J [1 ]
Humphrey, PPA [1 ]
机构
[1] Univ Cambridge, Dept Pharmacol, Glaxo Inst Appl Pharmacol, Cambridge CB2 1QJ, England
关键词
P2X; receptor; CHO-K1 cell line;
D O I
10.1038/sj.bjp.0702205
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
1 CHO-KI cells were examined for their cellular responses to the P2 receptor agonist, 2'- and 3'-O-(4-benzoylbenzoyl)-ATP (DbATP), and for the presence of mRNA for P2X receptors. 2 Reverse transcriptase-polymerase chain reactions, using primers directed against the rat P2X subunits, detected the presence of P2X, but not P2X(1)-P2X(6) subunits. 3 DbATP (EC(50 similar to)100 mu M) evoked non-desensitizing inward currents which reversed at similar to 0mV, suggesting activation of a non-selective cation channel. ATP also evoked inward currents but was less potent than DbATP. 4 DbATP also stimulated the accumulation of (45)calcium (Ca-45(2+)) and the DNA binding dye, YO-PRO-1, in CHO-K1 cells. Both responses were inhibited by NaCl and MgCl2. In 280 mM sucrose buffer, 45Ca(2+) accumulation was measurable within 10-20 s of agonist addition, whereas YO-PRO-1 accumulation was only detectable after 8 min. ATP and ATP gamma S were also agonists but were less potent than DbATP, while UTP, 2-methylthio ATP, ADP and alpha beta methylene ATP were inactive at concentrations up to 100 mu M. 5 DbATP increased lactate dehydrogenase release from CHO-K1 cells, suggesting cell lysis, although this effect was only pronounced after 60-90 min. 6 These data suggest that CHO-K1 cells express an endogenous P2X(7) receptor which can be activated by DbATP to cause a rapid inward current and accumulation of Ca-45(2+). Prolonged receptor activation results in a delayed, increased permeability to larger molecules such as YO-PRO-1 and ultimately leads to cell lysis. Importantly, the presence of an endogenous P2X(7) receptor should be considered when these cells are used to study recombinant P2X receptors.
引用
收藏
页码:1194 / 1201
页数:8
相关论文
共 21 条
  • [1] Properties of the pore-forming P2X(7) purinoceptor in mouse NTW8 microglial cells
    Chessell, IP
    Michel, AD
    Humphrey, PPA
    [J]. BRITISH JOURNAL OF PHARMACOLOGY, 1997, 121 (07) : 1429 - 1437
  • [2] ACTIVATION AND INHIBITION OF CALCIUM-DEPENDENT HISTAMINE-SECRETION BY ATP IONS APPLIED TO RAT MAST-CELLS
    COCKCROFT, S
    GOMPERTS, BD
    [J]. JOURNAL OF PHYSIOLOGY-LONDON, 1979, 296 (NOV): : 229 - 243
  • [3] THE ATP4- RECEPTOR OF RAT MAST-CELLS
    COCKCROFT, S
    GOMPERTS, BD
    [J]. BIOCHEMICAL JOURNAL, 1980, 188 (03) : 789 - 798
  • [4] INTERACTION OF ATP AND CALCIUM ON RAT MAST-CELL - EFFECT ON HISTAMINE-RELEASE
    DAHLQUIST, R
    DIAMANT, B
    [J]. ACTA PHARMACOLOGICA ET TOXICOLOGICA, 1974, 34 (05): : 368 - 384
  • [5] ERB L, 1990, J BIOL CHEM, V265, P7424
  • [6] A PATCH-CLAMP STUDY OF BOVINE CHROMAFFIN CELLS AND OF THEIR SENSITIVITY TO ACETYLCHOLINE
    FENWICK, EM
    MARTY, A
    NEHER, E
    [J]. JOURNAL OF PHYSIOLOGY-LONDON, 1982, 331 (OCT): : 577 - 597
  • [7] IMPROVED PATCH-CLAMP TECHNIQUES FOR HIGH-RESOLUTION CURRENT RECORDING FROM CELLS AND CELL-FREE MEMBRANE PATCHES
    HAMILL, OP
    MARTY, A
    NEHER, E
    SAKMANN, B
    SIGWORTH, FJ
    [J]. PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY, 1981, 391 (02): : 85 - 100
  • [8] HICKMAN SE, 1994, BLOOD, V84, P2452
  • [9] INCREASES IN INTRACELLULAR CALCIUM VIA ACTIVATION OF AN ENDOGENOUS P(2)-PURINOCEPTOR IN CULTURED CHO-K1 CELLS
    IREDALE, PA
    HILL, SJ
    [J]. BRITISH JOURNAL OF PHARMACOLOGY, 1993, 110 (04) : 1305 - 1310
  • [10] CONTROL OF MEMBRANE-PERMEABILITY BY EXTERNAL ATP IN MAMMALIAN-CELLS - ISOLATION OF AN ATP-RESISTANT VARIANT FROM CHINESE-HAMSTER OVARY CELLS
    KITAGAWA, T
    AKAMATSU, Y
    [J]. BIOCHIMICA ET BIOPHYSICA ACTA, 1986, 860 (02) : 185 - 193