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首页> 外文期刊>Cellular Signalling >Functional interactions between native Gs-coupled 5-HT receptors in HEK-293 cells and the heterologously expressed serotonin transporter
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Functional interactions between native Gs-coupled 5-HT receptors in HEK-293 cells and the heterologously expressed serotonin transporter

机译：HEK-293细胞中天然Gs偶联的5-HT受体与异源表达的血清素转运蛋白之间的功能相互作用

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In HEK-293 cells, serotonin (5-hydroxytryptamine, 5-HT) was found to induce cAMP production showing pharmacological characteristics consistent with the 5-HT7 receptor. The presence of 5-HT7 (and 5-HT6) receptor mRNA was confirmed by RT-PCR. Stable HEK-293 cell lines expressing either wild-type or haemagglutinin (HA)-tagged human 5-HT transporter (SERT) were selected and SERT function was confirmed using [3 H]5-HT transport. The presence of SERT caused a 10-fold reduction in the potency of 5-HT-induced cAMP production compared to control cells. Downstream signalling by 5-HT6/7 receptors could be detected as 5-HT-induced protein kinase A activation and phosphorylation of MAP kinase and CREB using phospho-specific antibodies. SERT inhibitors reversed the reduction in potency of 5-HT-induced cAMP production caused by the presence of SERT, resulting in a concentration-dependent left shift in EC50 values but also a progressive decrease in the maximal response. Thus, when antidepressants were used to block SERT activity, 5-HT receptor signalling was effectively clamped within a mid-range. (C) 2003 Elsevier Science Inc. All rights reserved. [References: 34]
机译：在HEK-293细胞中，发现5-羟色胺（5-羟色胺，5-HT）可诱导cAMP产生，显示出与5-HT7受体一致的药理特性。通过RT-PCR证实了5-HT7（和5-HT6）受体mRNA的存在。选择表达野生型或血凝素（HA）标签的人5-HT转运蛋白（SERT）的稳定的HEK-293细胞系，并使用[3 H] 5-HT转运确认SERT功能。与对照细胞相比，SERT的存在导致5-HT诱导的cAMP产生的效力降低了10倍。 5-HT6 / 7受体的下游信号传导可以检测为5-HT诱导的蛋白激酶A，使用磷酸特异性抗体激活并激活MAP激酶和CREB的磷酸化。 SERT抑制剂逆转了由于存在SERT而引起的5-HT诱导的cAMP产生的效力降低，从而导致EC50值与浓度有关的左移，但最大响应也逐渐降低。因此，当使用抗抑郁药来阻断SERT活性时，5-HT受体信号被有效地钳制在中间范围内。 （C）2003 Elsevier Science Inc.保留所有权利。 [参考：34]

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