Mutation of Three Residues in the Third Intracellular Loop of the Dopamine D2 Receptor Creates an Internalization-defective Receptor

Clayton, Cecilea C.; Donthamsetti, Prashant; Lambert, Nevin A.; Javitch, Jonathan A.; Neve, Kim A.

doi:10.1074/jbc.m114.605378

Cited by 36 publications

(66 citation statements)

References 41 publications

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“…RLuc8-fused receptor constructs (Urizar et al, 2011) were transfected with Venus-fused b arrestin-2 and GRK2, whose phosphorylation precedes and assists b arrestin recruitment (Evron et al, 2012). Although right-shifted compared with the G protein-related assays, the EC 50 values were in agreement with previous reports (Namkung et al, 2009;Clayton et al, 2014). Notably, consistent with the radioligand-binding and G protein-related experiments, the rank order of potency was D3R .…”

Section: Activation Of Dopamine D 2 -Like Receptors By Norepinephrinesupporting

confidence: 84%

Evidence for Noncanonical Neurotransmitter Activation: Norepinephrine as a Dopamine D₂-Like Receptor Agonist

Sánchez-Soto¹,

Bonifazi²,

Cai³

et al. 2016

Mol Pharmacol

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The Gai/o-coupled dopamine D 2 -like receptor family comprises three subtypes: the D 2 receptor (D2R), with short and long isoform variants (D2SR and D2LR), D 3 receptor (D3R), and D 4 receptor (D4R), with several polymorphic variants. The common overlap of norepinephrine innervation and D 2 -like receptor expression patterns prompts the question of a possible noncanonical action by norepinephrine. In fact, previous studies have suggested that norepinephrine can functionally interact with D4R. To our knowledge, significant interactions between norepinephrine and D2R or D3R receptors have not been demonstrated. By using radioligand binding and bioluminescent resonance energy transfer (BRET) assays in transfected cells, the present study attempted a careful comparison between dopamine and norepinephrine in their possible activation of all D 2 -like receptors, including the two D2R isoforms and the most common D4R polymorphic variants. Functional BRET assays included activation of G proteins with all Gai/o subunits, adenylyl cyclase inhibition, and b arrestin recruitment. Norepinephrine acted as a potent agonist for all D 2 -like receptor subtypes, with the general rank order of potency of D3R . D4R $ D2SR $ D2L. However, for both dopamine and norepinephrine, differences depended on the Gai/o protein subunit involved. The most striking differences were observed with Gai2, where the rank order of potencies for both dopamine and norepinephrine were D4R . D2SR 5 D2LR .. D3R. Furthermore the results do not support the existence of differences in the ability of dopamine and norepinephrine to activate different human D4R variants. The potency of norepinephrine for adrenergic a 2A receptor was only about 20-fold higher compared with D3R and D4R across the three functional assays.

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Section: Activation Of Dopamine D 2 -Like Receptors By Norepinephrinesupporting

confidence: 84%

Evidence for Noncanonical Neurotransmitter Activation: Norepinephrine as a Dopamine D₂-Like Receptor Agonist

Sánchez-Soto¹,

Bonifazi²,

Cai³

et al. 2016

Mol Pharmacol

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“…If a nocturnal animal is sampling odors while awake (when OB DA content is low), or while asleep during the day (when OB DA content is high), potent odors may activate dopaminergic interneurons so frequently that eventually the vesicular DA storage will be depleted. Additionally, constant DA release and binding can desensitize the presynaptic D 2 receptors on OSNs (Beaulieu et al, 2007; Clayton et al, 2014). Constant D 2 receptor activation would lead to phosphorylation of these receptors by G-protein receptor kinases, which would recruit the arrestin proteins, eventually leading to an internalization of the receptor (Beaulieu et al, 2007; Clayton et al, 2014).…”

Section: Olfactory Bulbmentioning

confidence: 99%

“…Additionally, constant DA release and binding can desensitize the presynaptic D 2 receptors on OSNs (Beaulieu et al, 2007; Clayton et al, 2014). Constant D 2 receptor activation would lead to phosphorylation of these receptors by G-protein receptor kinases, which would recruit the arrestin proteins, eventually leading to an internalization of the receptor (Beaulieu et al, 2007; Clayton et al, 2014). Thus, DA release may be gated by a low-pass filter, but DA activity, itself, may act as a high-pass filter, allowing for only those odors that surpass a threshold to be transmitted.…”

Section: Olfactory Bulbmentioning

confidence: 99%

Dopamine: A Modulator of Circadian Rhythms in the Central Nervous System

Korshunov

Blakemore

Trombley

2017

Front. Cell. Neurosci.

124

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Circadian rhythms are daily rhythms that regulate many biological processes – from gene transcription to behavior – and a disruption of these rhythms can lead to a myriad of health risks. Circadian rhythms are entrained by light, and their 24-h oscillation is maintained by a core molecular feedback loop composed of canonical circadian (“clock”) genes and proteins. Different modulators help to maintain the proper rhythmicity of these genes and proteins, and one emerging modulator is dopamine. Dopamine has been shown to have circadian-like activities in the retina, olfactory bulb, striatum, midbrain, and hypothalamus, where it regulates, and is regulated by, clock genes in some of these areas. Thus, it is likely that dopamine is essential to mechanisms that maintain proper rhythmicity of these five brain areas. This review discusses studies that showcase different dopaminergic mechanisms that may be involved with the regulation of these brain areas’ circadian rhythms. Mechanisms include how dopamine and dopamine receptor activity directly and indirectly influence clock genes and proteins, how dopamine’s interactions with gap junctions influence daily neuronal excitability, and how dopamine’s release and effects are gated by low- and high-pass filters. Because the dopamine neurons described in this review also release the inhibitory neurotransmitter GABA which influences clock protein expression in the retina, we discuss articles that explore how GABA may contribute to the actions of dopamine neurons on circadian rhythms. Finally, to understand how the loss of function of dopamine neurons could influence circadian rhythms, we review studies linking the neurodegenerative disease Parkinson’s Disease to disruptions of circadian rhythms in these five brain areas. The purpose of this review is to summarize growing evidence that dopamine is involved in regulating circadian rhythms, either directly or indirectly, in the brain areas discussed here. An appreciation of the growing evidence of dopamine’s influence on circadian rhythms may lead to new treatments including pharmacological agents directed at alleviating the various symptoms of circadian rhythm disruption.

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“…Receptor and arrestin biosensors can be transiently or stably transfected, and can be co-expressed with G protein receptor kinases (GRKs) to enhance agonist-dependent phosphorylation of the receptor and subsequent arrestin recruitment (Clayton et al, 2014). This protocol can be applied to a wide variety of GPCRs.…”

Section: Introductionmentioning

confidence: 99%

“…Therefore, we have developed a novel BRET based-assay to overcome this problem (Clayton et al, 2014). Rather than being directly fused to the receptor, the BRET sensor is instead fused to an unrelated plasma membrane marker.…”

Section: Introductionmentioning

confidence: 99%

Using Bioluminescence Resonance Energy Transfer (BRET) to Characterize Agonist‐Induced Arrestin Recruitment to Modified and Unmodified G Protein‐Coupled Receptors

et al. 2015

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G protein-coupled receptors (GPCRs) represent ~25% of current drug targets. Ligand binding to these receptors activates G proteins and arrestins, which are involved in differential signaling pathways. Functionally selective or biased ligands activate one of these two pathways and may be superior medications for certain diseases states. The identification of these ligands requires robust drug screening assays for both G protein and arrestin activity. Here we describe in detail the technical aspects of two bioluminescence resonance energy (BRET)-based assays that can be used to monitor arrestin recruitment to GPCRs. One assay requires modification of GPCRs by fusion to a BRET donor or acceptor moiety, whereas the other can detect recruitment of arrestin to unmodified GPCRs.

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Mutation of Three Residues in the Third Intracellular Loop of the Dopamine D2 Receptor Creates an Internalization-defective Receptor

Cited by 36 publications

References 41 publications

Evidence for Noncanonical Neurotransmitter Activation: Norepinephrine as a Dopamine D₂-Like Receptor Agonist

Evidence for Noncanonical Neurotransmitter Activation: Norepinephrine as a Dopamine D₂-Like Receptor Agonist

Dopamine: A Modulator of Circadian Rhythms in the Central Nervous System

Using Bioluminescence Resonance Energy Transfer (BRET) to Characterize Agonist‐Induced Arrestin Recruitment to Modified and Unmodified G Protein‐Coupled Receptors

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Mutation of Three Residues in the Third Intracellular Loop of the Dopamine D2 Receptor Creates an Internalization-defective Receptor

Cited by 36 publications

References 41 publications

Evidence for Noncanonical Neurotransmitter Activation: Norepinephrine as a Dopamine D2-Like Receptor Agonist

Evidence for Noncanonical Neurotransmitter Activation: Norepinephrine as a Dopamine D2-Like Receptor Agonist

Dopamine: A Modulator of Circadian Rhythms in the Central Nervous System

Using Bioluminescence Resonance Energy Transfer (BRET) to Characterize Agonist‐Induced Arrestin Recruitment to Modified and Unmodified G Protein‐Coupled Receptors

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Product

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Evidence for Noncanonical Neurotransmitter Activation: Norepinephrine as a Dopamine D₂-Like Receptor Agonist

Evidence for Noncanonical Neurotransmitter Activation: Norepinephrine as a Dopamine D₂-Like Receptor Agonist