The organic cation transporter 2 regulates dopamine D1 receptor signaling at the Golgi apparatus.
The organic cation transporter 2 regulates dopamine D1 receptor signaling at the Golgi apparatus.
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DOI：
10.7554/elife.75468
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发表时间：
2022-04-25
期刊：
影响因子：
7.7
通讯作者：
Irannejad, Roshanak
中科院分区：
文献类型：
--
作者：
Puri, Natasha M.;Romano, Giovanna R.;Lin, Ting-Yu;Mai, Quynh N.;Irannejad, Roshanak
关键词：
多巴胺是大脑和肾脏中的一种关键的儿茶酚胺，它参与许多生理功能，如运动、认知、情感、内分泌调节和肾功能。作为一种膜外激素和神经递质，多巴胺被认为仅在质膜上通过结合和激活多巴胺受体（G蛋白偶联受体（GPCR）家族成员）来传递信号。在这项研究中，我们首次利用基于纳米体的新型生物传感器证明了多巴胺D1受体（D1DR）不仅在质膜上起作用，而且在配体存在的情况下在高尔基体上被激活。D1DR是大脑和肾脏中多巴胺能信号传导的主要介质。我们提供的证据表明，D1DR的高尔基池的激活依赖于有机阳离子转运体2 (OCT2)，一种多巴胺转运体，这就解释了膜外多巴胺如何进入D1DR的亚细胞池。我们进一步证明多巴胺激活小鼠纹状体中棘神经元Golgi-D1DR，这种活性依赖于OCT2功能。我们还介绍了一种新的方法，通过使用基于纳米体的化学招募系统，通过抑制Gαs偶联来选择性地询问区隔化的D1DR信号。使用这种策略，我们发现高尔基定位的d1dr调节cAMP的产生并介导局部蛋白激酶A的激活。综上所述，我们的数据表明，空间分区的信号集线器是D1DR信号的先前未被重视的调节方面。我们的数据为转运体在调节亚细胞GPCR活性中的作用提供了进一步的证据。
Dopamine is a key catecholamine in the brain and kidney, where it is involved in a number of physiological functions such as locomotion, cognition, emotion, endocrine regulation, and renal function. As a membrane-impermeant hormone and neurotransmitter, dopamine is thought to signal by binding and activating dopamine receptors, members of the G protein coupled receptor (GPCR) family, only on the plasma membrane. Here, using novel nanobody-based biosensors, we demonstrate for the first time that the dopamine D1 receptor (D1DR), the primary mediator of dopaminergic signaling in the brain and kidney, not only functions on the plasma membrane but becomes activated at the Golgi apparatus in the presence of its ligand. We present evidence that activation of the Golgi pool of D1DR is dependent on organic cation transporter 2 (OCT2), a dopamine transporter, providing an explanation for how the membrane-impermeant dopamine accesses subcellular pools of D1DR. We further demonstrate that dopamine activates Golgi-D1DR in murine striatal medium spiny neurons, and this activity depends on OCT2 function. We also introduce a new approach to selectively interrogate compartmentalized D1DR signaling by inhibiting Gαs coupling using a nanobody-based chemical recruitment system. Using this strategy, we show that Golgi-localized D1DRs regulate cAMP production and mediate local protein kinase A activation. Together, our data suggest that spatially compartmentalized signaling hubs are previously unappreciated regulatory aspects of D1DR signaling. Our data provide further evidence for the role of transporters in regulating subcellular GPCR activity.