The organic cation transporter 2 regulates dopamine D1 receptor signaling at the Golgi apparatus
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 a...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2022-04-01
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| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/75468 |
| _version_ | 1818026268362276864 |
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| author | Natasha M Puri Giovanna R Romano Ting-Yu Lin Quynh N Mai Roshanak Irannejad |
| author_facet | Natasha M Puri Giovanna R Romano Ting-Yu Lin Quynh N Mai Roshanak Irannejad |
| author_sort | Natasha M Puri |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-10T04:29:18Z |
| format | Article |
| id | doaj.art-596732ebca9545fa88cd4c34dcf70187 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-12-10T04:29:18Z |
| publishDate | 2022-04-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-596732ebca9545fa88cd4c34dcf701872022-12-22T02:02:11ZengeLife Sciences Publications LtdeLife2050-084X2022-04-011110.7554/eLife.75468The organic cation transporter 2 regulates dopamine D1 receptor signaling at the Golgi apparatusNatasha M Puri0https://orcid.org/0000-0002-4290-4361Giovanna R Romano1Ting-Yu Lin2Quynh N Mai3https://orcid.org/0000-0002-6199-2096Roshanak Irannejad4https://orcid.org/0000-0001-8702-2285Department of Biochemistry & Biophysics, University of California, San Francisco, San Francisco, United StatesDepartment of Biochemistry & Biophysics, University of California, San Francisco, San Francisco, United States; Biochemistry Department, Weill Cornell Medicine, New York, United StatesCardiovascular Research Institute, University of California, San Francisco, San Francisco, United StatesCardiovascular Research Institute, University of California, San Francisco, San Francisco, United StatesDepartment of Biochemistry & Biophysics, University of California, San Francisco, San Francisco, United States; Cardiovascular Research Institute, University of California, San Francisco, San Francisco, United StatesDopamine 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.https://elifesciences.org/articles/75468D1 dopamine receptorOCT2 transporterGolgi apparatuscAMPstriatal neuronscompartmentalized signaling |
| spellingShingle | Natasha M Puri Giovanna R Romano Ting-Yu Lin Quynh N Mai Roshanak Irannejad The organic cation transporter 2 regulates dopamine D1 receptor signaling at the Golgi apparatus eLife D1 dopamine receptor OCT2 transporter Golgi apparatus cAMP striatal neurons compartmentalized signaling |
| title | The organic cation transporter 2 regulates dopamine D1 receptor signaling at the Golgi apparatus |
| title_full | The organic cation transporter 2 regulates dopamine D1 receptor signaling at the Golgi apparatus |
| title_fullStr | The organic cation transporter 2 regulates dopamine D1 receptor signaling at the Golgi apparatus |
| title_full_unstemmed | The organic cation transporter 2 regulates dopamine D1 receptor signaling at the Golgi apparatus |
| title_short | The organic cation transporter 2 regulates dopamine D1 receptor signaling at the Golgi apparatus |
| title_sort | organic cation transporter 2 regulates dopamine d1 receptor signaling at the golgi apparatus |
| topic | D1 dopamine receptor OCT2 transporter Golgi apparatus cAMP striatal neurons compartmentalized signaling |
| url | https://elifesciences.org/articles/75468 |
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