Modulation of dopaminergic neurotransmission by ADGRL3, an adhesion GPCR associated with ADHD susceptibility

ADGRL3（一种与 ADHD 易感性相关的粘附 GPCR）对多巴胺能神经传递的调节

• 批准号: 9350414
• 负责人: Jonathan A Javitch
• 金额: $ 20.25万
• 依托单位: NEW YORK STATE PSYCHIATRIC INSTITUTE DBA RESEARCH FOUNDATION FOR MENTAL HYGIENE, INC
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-09 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9350414
• 关键词: Adhesions; Adhesives; Agonist; Animals; Architecture; Attention deficit hyperactivity disorder; Back; Behavioral; Behavioral Model; Binding; Biological Assay; Cell membrane; Cognition; Complex; Corpus striatum structure; Disease; Dopamine; Drosophila genus; Family; Fibronectins; Functional disorder; Future; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; G-substrate; GTP-Binding Proteins; Genes; Genetic; Genetic Polymorphism; Goals; Hyperactive behavior; Image; In Vitro; Integral Membrane Protein; Knockout Mice; Ligand Binding; Ligands; Magnetism; Mechanical Stress; Mechanics; Mediating; Mental Health; Morphology; Motor Activity; Mus; Mutation; N-terminal; Orphan; Pathway interactions; Patients; Peptide Hydrolases; Pharmacotherapy; Phenotype; Physiological; Predisposition; Property; Protein Array; Proteins; Proteolysis; Risk; Role; Schizophrenia; Signal Pathway; Signal Transduction; Site; Synapses; Tertiary Protein Structure; Testing; Therapeutic; Transmembrane Domain; Zebrafish; addiction; alpha-latrotoxin receptor; dopaminergic neuron; drug development; extracellular; improved; in vitro activity; in vivo; innovation; interdisciplinary approach; leucine-rich repeat protein; loss of function; mechanical force; neuropsychiatric disorder; neurotransmission; novel; receptor; screening; sensor; therapy development; tool

Project Summary Adhesion GPCRs (aGPCRs) form the second largest, yet most enigmatic class of the GPCR superfamily. Recent genetic findings show that polymorphisms in the gene encoding the adhesion G protein-coupled receptor (aGPCR) latrophilin 3 (ADGRL3) are strongly associated with an increased risk of attention deficit hyperactivity disorder (ADHD). Remarkably, in fruit flies, zebrafish and mice, disruption of ADGRL3 expression enhanced locomotor activity. In adgrl3 null mice, striatal dopamine levels were shown to be increased, suggesting that the cross-species hyperactive phenotype may be mediated through enhanced dopamine signaling. Thus, ADGRL3 represents a novel target for the development of drug treatments for ADHD and other neuropsychiatric disorders that involve dopamine dysregulation, such as schizophrenia and addiction. However, ADGRL3 remains orphan with respect to the identity of its agonists and signaling properties, and its role in dopamine neurotransmission is poorly understood. Like other aGPCRs, ADGRL3 contains a 7 transmembrane domain and an extracellular N-terminal architecture comprising an array of protein domains suitable for adhesive interactions with protein ligands. In the current proposal we aim to characterize the basic signaling properties of ADGRL3 and how they can be modulated by adhesive protein ligands. For this purpose we will use a comprehensive suite of GPCR signaling assays combined with an innovative magnetic force assay to mimic the mechanical force potentially exerted in binding of trans-synaptic ligands that function to maintain and modulate synapse morphology and organization. In parallel, we will use a combination of genetic strategies together with imaging and behavioral approaches to characterize the role of ADGRL3 in dopamine neurotransmission in vivo. This multidisciplinary approach will also serve as a platform for future studies aimed at testing the functional impact of disease mutations in ADGRL3 and in screening for modulators of ADGRL3 signaling. To achieve these goals we propose the following specific aims: 1) To identify the G protein pathways controlled by ADGRL3 and to establish the role of the N terminus in signaling and in regulating dopamine neurotransmission using (a) in vitro signaling assays and strategic ADGRL3 mutations, (b) an innovative magnetic tweezer assay to evaluate the influence of mechanical force on the activity of ADGRL3 and (c) a Drosophila behavioral model to characterize the localization and function of ADGRL3 in dopaminergic neurons in vivo. 2) To identify the mechanism by which trans-synaptic ligands modulate ADGRL3 signaling and dopamine neurotransmission by determining the effect of teneurin-1 and other ligands on ADGRL3 signaling using the in vitro and in vivo approaches described above.

项目摘要 粘附GPCR（AGPCR）构成了GPCR超家族的第二大但最神秘的类别。 最近的遗传发现表明，编码粘附G蛋白偶联的基因中的多态性 受体（AGPCR）Latrophilin 3（ADGRL3）与注意力不足的风险增加密切相关 多动症（ADHD）。值得注意的是，在果蝇，斑马鱼和小鼠中，ADGRL3表达的破坏 增强的运动活性。在ADGRL3无效小鼠中，纹状体多巴胺水平显示出增加， 表明跨物种过度活跃表型可以通过增强的多巴胺介导 信号。因此，ADGRL3代表了开发多动症药物治疗和 其他涉及多巴胺失调的神经精神疾病，例如精神分裂症和成瘾。 但是，ADGRL3关于其激动剂和信号特性的身份及其身份仍然是孤儿 在多巴胺神经传递中的作用知之甚少。像其他AGPCR一样，ADGRL3包含一个7 跨膜结构域和一个包括蛋白质结构群的细胞外N末端结构 适用于与蛋白质配体的粘合剂相互作用。在当前的建议中，我们旨在表征基本 ADGRL3的信号传导特性以及如何通过粘合蛋白配体调节它们。为此目的 我们将使用全面的GPCR信号测定套件，并结合创新的磁力 测定模拟可能施加的机械力，可能在跨突触配体的结合中发挥作用 维护和调节突触的形态和组织。同时，我们将使用遗传的组合 策略以及成像和行为方法来表征ADGRL3在多巴胺中的作用 体内神经传递。这种多学科的方法还将成为未来研究的平台 测试疾病突变在ADGRL3和筛选ADGRL3的功能影响时 信号。为了实现这些目标，我们提出以下特定目的：1）确定G蛋白 由ADGRL3控制的途径，并确定n末端在信号传导中的作用 使用（a）体外信号传导和战略ADGRL3调节多巴胺神经传递 突变，（b）一种创新的磁性镊子测定，以评估机械力对 ADGRL3和（C）果蝇行为模型的活性，以表征 在体内多巴胺能神经元中的ADGRL3。 2）确定反式突触配体的机制 通过确定Teneurin-1和 使用上述体外和体内方法进行ADGRL3信号传导上的其他配体。

项目成果

Publisher Correction: G12/13 is activated by acute tethered agonist exposure in the adhesion GPCR ADGRL3.

出版商更正：G12/13 是由粘附 GPCR ADGRL3 中的急性束缚激动剂暴露激活的。

• DOI: 10.1038/s41589-020-0649-z
• 发表时间: 2020
• 期刊: Nature chemical biology
• 影响因子: 14.8
• 作者: Mathiasen,Signe;Palmisano,Tiago;Perry,NicoleA;Stoveken,HannahM;Vizurraga,Alex;McEwen,DykeP;Okashah,Najeah;Langenhan,Tobias;Inoue,Asuka;Lambert,NevinA;Tall,GregoryG;Javitch,JonathanA
• 通讯作者: Javitch,JonathanA