CHARACTERIZATION OF THE 5-HYDROXYTRYPTAMINE(2A) RECEPTOR-ACTIVATED CASCADE IN RAT C6 GLIOMA-CELLS

被引:27
作者
ELLIOTT, JM [1 ]
NEWBERRY, NR [1 ]
CHOLEWINSKI, AJ [1 ]
BARTRUP, JT [1 ]
BRIDDON, SJ [1 ]
CAREY, JE [1 ]
FLANIGAN, TP [1 ]
NEWTON, RA [1 ]
PHIPPS, SL [1 ]
REAVLEY, AC [1 ]
SMITH, C [1 ]
WIGMORE, M [1 ]
GRAHAMESMITH, DG [1 ]
LESLIE, RA [1 ]
机构
[1] SMITHKLINE BEECHAM PHARMACEUT,DEPT BIOTECHNOL,HARLOW CM19 5AW,ESSEX,ENGLAND
基金
英国惠康基金;
关键词
CA2+; K+ CHANNEL; H-3] KETANSERIN; DESENSITIZATION; DOWN-REGULATION; DIFFERENTIATION;
D O I
10.1016/0306-4522(95)00323-B
中图分类号
Q189 [神经科学];
学科分类号
071006 ;
摘要
We have investigated the identity and intracellular cascade of responses resulting from activation of the endogenous 5-hydroxytryptamine receptor in the C6 rat glioma cell line. Sequence analysis of reverse transcription-polymerase chain reaction products derived from C6 glioma cell messenger RNA revealed complete homology with a portion of the rat 5-hydroxytryptamine(2A) receptor. The binding of [H-3]ketanserin to cell membranes demonstrated a significant correlation with the 5-hydroxytryptamine(2A) receptor in rat frontal cortex. On intact cells, 5-hydroxytryptamine stimulated a concentration-dependent increase in phosphatidyl inositide turnover and intracellular [Ca2+] mediated by 5-hydroxytryptamine(2A) receptors. In whole-cell patch-clamp recordings, 5-hydroxytryptamine induced an outward current mediated predominantly by K+ ions (reversal potential = -80 mV). Using caged molecules containing Ca2+ or inositol 1,4,5-trisphosphate in the patch electrode solution, we found that rapid photolytic release of Ca2+ and particularly inositol 1,4,5-trisphosphate within the cytosol induced an outward current with characteristics similar to those seen after application of 5-hydroxytryptamine. Comparison between differentiated and undifferentiated cells revealed significantly higher receptor density and maximal phosphoinositide response to 5-hydroxytryptamine in undifferentiated cells but the associated rise in [Ca2+](i) and activation of an outward current was observed more frequently in differentiated cells. Prolonged exposure of the cells to 5-hydroxytryptamine led to a decrease in all responses and to the down-regulation of receptor number. We conclude that the rat C6 glioma cell expresses a 5-hydroxytryptamine(2A) receptor identical to that found in rat brain and that stimulation of the receptor in C6 cells leads to the activation of Ca2+ activated K+ channels via phosphoinositide hydrolysis and subsequent rise in cytosolic Ca2+ ion concentration. However, the contrasting effects of differentiation on receptor number and phosphoinositide response to 5-hydroxytryptamine compared to Ca2+ release and conductance change indicate that a complex relationship exists between the component parts of the receptor-activated cascade.
引用
收藏
页码:1119 / 1131
页数:13
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