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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >AGONIST-INDUCED DESENSITIZATION OF DOPAMINE D1 RECEPTOR-STIMULATED ADENYLYL CYCLASE ACTIVITY IS TEMPORALLY AND BIOCHEMICALLY SEPARATED FROM D1 RECEPTOR INTERNALIZATION
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AGONIST-INDUCED DESENSITIZATION OF DOPAMINE D1 RECEPTOR-STIMULATED ADENYLYL CYCLASE ACTIVITY IS TEMPORALLY AND BIOCHEMICALLY SEPARATED FROM D1 RECEPTOR INTERNALIZATION

机译：激动剂诱导的多巴胺D1受体刺激的腺苷酸环化酶活性的去化是暂时的，并且是从D1受体的内化过程中生物化学分离的

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摘要

The regulation of the dopamine D1 receptor was investigated by using c-myc epitope-tagged D1 receptors expressed in Sf9 (fall armyworm ovary) cells. Treatment of D1 receptors with 10 mu M dopamine for 15 min led to a loss of the dopamine-detected high-affinity state of the receptor accompanying a 40% reduction in the ability of the receptor to mediate maximal dopamine stimulation of adenylyl cyclase activity. After 60 min of agonist exposure, 45 min after the occurrence of desensitization, 28% of the cell surface receptors were internalized into an intracellular light vesicular membrane fraction as determined by radioligand binding and supported by photoaffinity labeling, immunocytochemical staining, and immunoblot analysis. Pretreatment of cells with concanavalin A or sucrose completely blocked agonist-induced D1 receptor internalization without preventing agonist-induced desensitization, indicating a biochemical separation of these processes. Collectively, these findings indicate that the desensitization of D1 receptor-coupled adenylyl cyclase activity and D1 receptor internalization are temporally and biochemically distinct mechanisms regulating D1 receptor function following agonist activation. [References: 17]
机译：多巴胺D1受体的调节是通过使用在Sf9（秋天粘虫卵巢）细胞中表达的c-myc表位标记的D1受体来研究的。用10μM多巴胺处理D1受体15分钟会导致多巴胺检测到的受体高亲和力状态丧失，同时受体介导最大程度的多巴胺刺激腺苷酸环化酶活性的能力降低40％。激动剂暴露60分钟后，脱敏发生45分钟后，通过放射性配体结合确定并通过光亲和标记，免疫细胞化学染色和免疫印迹分析支持，将28％的细胞表面受体内化到细胞内轻囊泡膜部分中。用伴刀豆球蛋白A或蔗糖预处理的细胞完全阻断了激动剂诱导的D1受体内在化，而没有阻止激动剂诱导的脱敏，表明这些过程的生化分离。总的来说，这些发现表明D1受体偶联的腺苷酸环化酶活性的脱敏和D1受体内在化是激动剂激活后在时间和生化上不同的调节D1受体功能的机制。 [参考：17]

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