Deciphering Caenorhabditis elegans molecular nociception using mass spectrometry-based proteomics and network biology

使用基于质谱的蛋白质组学和网络生物学破译秀丽隐杆线虫分子伤害感受

• 批准号: RGPIN-2015-05071
• 负责人: Beaudry, Francis
• 金额: $ 2.04万
• 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: 2018
• 资助国家: 加拿大
• 起止时间: 2018-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=644895
• 关键词: Deciphering; Caenorhabditis; elegans; molecular; nociception

Caenorhabditis elegans (C.elegans) is a powerful animal model system for functional genomics relevant to mammalian biology and it is extensively used to study nociceptive behavior. Interestingly, adult C.elegans consists of 959 cells, of which 302 are neurons, which make this model attractive to study nociception at the physiological and molecular levels. Transient receptor potential (TRP) channels have a central role in the processing of nociceptive chemical and thermal stimuli. In mammals, the activation of TRP channels triggers the propagation of sensory information back to the central nervous system, which leads to pain sensation. Pro-inflammatory sensory neuropeptides, such as tachykinins are released from these activated nerve endings resulting in neurogenic inflammation. At the spinal level, neuropeptides play a central role in pain transmission and in the synaptic chemistry. The C.elegans genome encodes several TRP channels covering all of the seven TRP subfamilies including TRPV analogs (e.g. 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. Moreover, recent studies revealed that neuropeptides from the MSFamide family (NLPs) and FMRamide-related peptides (FLPs) share functional similarities with mammal neuropeptides but very few have been thoroughly characterized. Neuropeptides are involved in the modulation of essentially all behaviors including locomotion, mechano- and chemosensation. Neuropeptides are acting as neuromodulators and as fast neurotransmitters. The broad existence of these neuropeptides in nematodes suggests a fundamental role of neuropeptidergic signalling in C.elegans but the mechanistic knowledge remains very limited. The objectives of the proposed research will be to (1) Quantify nociceptive behaviors of wild type and targeted mutant (e.g. OSM-9, OCR1-4) C.elegans associated with thermal and chemical stimuli; (2) Characterize C.elegans by mass spectrometry using proteomic bottom-up approaches and SILAC normalization; (3) Generate comprehensive bioinformatics analyses (e.g. Ingenuity Pathway Analysis, GeneMANIA) using the proteomic data to identify relationships, mechanisms, functions, biological networks and pathways of relevance. We believed it will provide significant insight in the molecular events implicated in C.elegans nociceptive behaviors and chemical synapses. A better comprehension of these mechanisms in C.elegans will transform our powers of investigation, allowing us, and others, to pursue more comprehensive study of nociception at the molecular level.**

秀丽隐杆线虫是研究哺乳动物生物学功能基因组学的重要动物模型系统，广泛用于研究伤害性行为。有趣的是，成年秀丽隐杆线虫由959个细胞组成，其中302个是神经元，这使得该模型在生理和分子水平上研究伤害感受具有吸引力。瞬时受体电位（TRP）通道在伤害性化学和热刺激的处理中具有中心作用。在哺乳动物中，TRP通道的激活触发感觉信息返回中枢神经系统的传播，这导致疼痛感觉。促炎性感觉神经肽，如速激肽从这些激活的神经末梢释放，导致神经源性炎症。在脊髓水平，神经肽在疼痛传递和突触化学中起核心作用。秀丽隐杆线虫基因组编码几个TRP通道，覆盖所有七个TRP亚家族，包括TRPV类似物（例如OSM-9和OCR 1 -4）。最近已经确定秀丽隐杆线虫TRP通道与行为和生理过程相关，包括感觉转导。许多秀丽隐杆线虫TRP通道与它们的脊椎动物对应物共享相似的激活和调节机制。此外，最近的研究表明，神经肽从MSFamide家庭（NLP）和FMR酰胺相关的肽（FLP）共享功能相似性与哺乳动物的神经肽，但很少有被彻底的特点。神经肽参与基本上所有行为的调节，包括运动、机械感觉和化学感觉。神经肽作为神经调质和快速神经递质发挥作用。这些神经肽在线虫中的广泛存在表明神经肽能信号在秀丽隐杆线虫中的基本作用，但机制知识仍然非常有限。本研究的主要目的是：（1）定量野生型和靶向突变体的伤害性行为（2）使用蛋白质组学自下而上方法和SILAC标准化通过质谱法表征秀丽隐杆线虫;（3）使用蛋白质组数据生成全面的生物信息学分析（例如，Inflamity Pathway Analysis，GeneMANIA），以确定相关的关系，机制，功能，生物网络和途径。我们相信这将为线虫伤害性行为和化学突触的分子机制研究提供重要的线索。更好地理解秀丽隐杆线虫的这些机制将改变我们的研究能力，使我们和其他人能够在分子水平上对伤害感受进行更全面的研究。