Devlin Mark G, Smith Nicola J, Ryan Olivia M, Guida Elizabeth, Sexton Patrick M, Christopoulos Arthur
Department of Pharmacology, University of Melbourne, Parkville, 3010 Victoria, Australia.
Eur J Pharmacol. 2004 Sep 13;498(1-3):59-69. doi: 10.1016/j.ejphar.2004.07.102.
The effect of ligand pretreatment on human 5-hydroxytryptamine2C (5-HT2C) receptors was examined in CHO cells expressing high (CHO-1C7; 67+/-3 pmol/mg) or low (CHO-1C19; 72+/-10 fmol/mg) levels of the receptor. Seventy-two hours pretreatment of CHO-1C7 cells with various ligands did not affect receptor expression. Pretreatment with inverse agonists enhanced 5-HT-mediated inositol phosphate accumulation with no change in constitutive receptor activity. The enhanced agonist responsiveness was inversely correlated with the intrinsic activity of the pretreatment ligand. Seventy-two hours of pretreatment with the weak agonist, 5-methoxygramine, caused an elevation in constitutive activity but no alteration in 5-HT-mediated signaling. In CHO-1C19 cells, 24 but not 72 h of pretreatment with the inverse agonist mianserin enhanced 5-HT-mediated signaling, with no effect on basal signaling; pretreatment with 5-methoxygramine had no significant effect. These findings highlight differences in the pattern of chronic regulation of 5HT2C receptor signaling between high and low receptor expression levels in a common cellular background.
在表达高水平(CHO-1C7;67±3 pmol/mg)或低水平(CHO-1C19;72±10 fmol/mg)人5-羟色胺2C(5-HT2C)受体的CHO细胞中,研究了配体预处理对该受体的影响。用各种配体对CHO-1C7细胞进行72小时预处理,不影响受体表达。用反向激动剂预处理可增强5-羟色胺介导的肌醇磷酸积累,而组成型受体活性无变化。增强的激动剂反应性与预处理配体的内在活性呈负相关。用弱激动剂5-甲氧基色胺预处理72小时,导致组成型活性升高,但5-羟色胺介导的信号传导无改变。在CHO-1C19细胞中,用反向激动剂米安色林预处理24小时而非72小时可增强5-羟色胺介导的信号传导,对基础信号传导无影响;用5-甲氧基色胺预处理无显著影响。这些发现突出了在共同细胞背景下,高受体表达水平和低受体表达水平之间5HT2C受体信号慢性调节模式的差异。