Decrypting nociceptive signaling pathways in Caenorhabditis elegans using behavioral analysis and mass spectrometry-based proteomics

使用行为分析和基于质谱的蛋白质组学解密秀丽隐杆线虫的伤害性信号通路

• 批准号: RGPIN-2020-05228
• 负责人: Beaudry, Francis
• 金额: $ 3.06万
• 依托单位: Université de Montréal
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2020
• 资助国家: 加拿大
• 起止时间: 2020-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=717571
• 关键词: Decrypting; nociceptive; signaling; pathways; Caenorhabditis

My NSERC Discovery research program is dedicated to understanding the molecular processes associated with nociceptive behaviors, the neurochemistry of synaptic transmission and identify targets that can regulate nociceptive processing. The overall goal of this proposal will be to 1) characterize the effects of endocannabinoids and cannabinoids on nocifensive response of Caenorhabditis elegans (C. elegans) to noxious heat and compared with vanilloids, 2) characterize the interplay between cannabinoid and vanilloid systems using specific C. elegans mutants, pharmacological manipulations and proteomics; 3) identify the molecular target of endocannabinoids, cannabinoids and vanilloids in C. elegans using a variant of the thermal shift assays used to study thermal stabilization of proteins upon ligand binding combined with an unbiased, proteome-wide approach for the identification of specific-binding protein targets. C. elegans is a powerful experimental model system relevant to mammalian physiology. C. elegans consists of 959 cells, of which 302 are neurons, which makes this model attractive to study nociception, the sensory nervous system's responsible for triggering a response to harmful stimuli. Nociceptors are sensory receptors specialized in detecting potential harm, including temperature, mechanical, and chemical. Transient receptor potential (TRP) channels are essential detectors and transducers of harmful stimuli. C. elegans genome encodes several TRP channels including transient receptor potential cation channel subfamily V (TRPV or vanilloid receptor) orthologs (OSM-9 and OCR1-4). It has been recently established that C. elegans TRP channels are associated with behavioral and physiological processes, including sensory transduction. Many C. elegans TRP channels share similar activation and regulatory mechanisms with their vertebrate counterparts. The endocannabinoid system is involved in modulation of nociception and recent evidence suggests interactions with the vanilloid systems. Many studies identified components of a C. elegans endocannabinoid system including numerous endocannabinoid ligands. Thus, we believe that C. elegans expresses receptors that act as functional homologs of mammalian cannabinoid receptors and is an ideal experimental model to study new cannabinoid signaling pathways and their interplay with the vanilloid systems. In addition to the two main cannabinoid receptors (CB1 and CB2), there is growing evidence that there are other, not yet identified, cannabinoid receptors in mammals. This research proposal will have a significant impact on the production of knowledge fundamental to characterize heat activated signaling pathways mediated by vanilloid receptors and their interaction(s) with the cannabinoid system components. This is a fundamental step to ensure translational success of experimental models. Moreover, this comprehensive study may lead to the discovery of a novel cannabinoid receptor.

我的NSERC Discovery研究项目致力于了解与伤害性行为相关的分子过程，突触传递的神经化学，并确定可以调节伤害性感受处理的靶点。 这项建议的总体目标将是1)表征内源性大麻素和大麻素对线虫对有害热的伤害反应的影响，并与香草素进行比较；2)使用特定的线虫突变体、药理操作和蛋白质组学来表征大麻素和香草素系统之间的相互作用；3)使用一种用于研究蛋白质对连接和结合的热稳定性的变种热位移分析方法来确定线虫中内源性大麻素、大麻素和香草素的分子靶标。 线虫是与哺乳动物生理学相关的一个强大的实验模型系统。线虫由959个细胞组成，其中302个是神经元，这使得这个模型对研究伤害感很有吸引力，伤害感是感官神经系统负责触发对有害刺激的反应。伤害性感受器是专门用于检测潜在伤害的感官受体，包括温度、机械和化学物质。瞬时感受器电位(Trp)通道是有害刺激的重要检测器和传导器。线虫基因组编码多种色氨酸通道，包括瞬时受体潜势阳离子通道亚家族V(TRPV或香草样受体)同源基因(OSM-9和OCR1-4)。最近发现线虫的Trp通道与包括感觉转导在内的行为和生理过程有关。许多线虫的Trp通道与脊椎动物有相似的激活和调节机制。内源性大麻素系统参与了伤害性感觉的调节，最近的证据表明它与香草素系统相互作用。许多研究确定了线虫内源性大麻素系统的成分，包括大量的内源性大麻素配体。因此，我们认为线虫表达的受体是哺乳动物大麻素受体的功能同源物，是研究新的大麻素信号通路及其与香草素系统相互作用的理想实验模型。除了两种主要的大麻素受体(CB1和CB2)外，越来越多的证据表明，哺乳动物中还有其他尚未鉴定的大麻素受体。 这项研究提案将对知识的产生产生重大影响，这些知识是表征香草素受体介导的热激活信号通路及其与大麻素系统成分的相互作用的基础(S)。这是确保实验模型翻译成功的根本步骤。此外，这项全面的研究可能导致发现一种新的大麻素受体。