Investigating novel mechanical receptors in the sensory nervous system

哺乳动物如何察觉钝力或尖锐物体的有害性质？身体是如何防御性地增加受伤或发炎组织的敏感性，使原本无害的动作和触摸感到疼痛的？作为衰老不可避免的后果，后一个过程是如何随着时间的推移而出现问题，造成沉重的慢性疼痛的？要回答这些问题，我们必须看看感觉神经。这些细胞为我们的器官（包括皮肤）提供检测环境物理和化学性质信息的能力。它们是通过一种叫做受体的蛋白质分子来做到这一点的，这种蛋白质分子可以检测到特定类型的环境线索。在过去的十年里，我们已经了解了很多关于感知热、冷和轻触的感受器的知识。然而，那些导致触摸疼痛的基因仍然难以捉摸。发现并描述这些分子会带来很高的回报。除了进一步加深我们对感觉生理学的了解，这些将为开发新的、更有效的、副作用更小的疼痛缓解疗法提供潜在的目标。该项目将使用最先进的功能基因组学和电生理学技术来识别和研究这些蛋白质。该项目的总体目标是在感觉神经系统中识别新的分子受体，特别关注那些有害的机械刺激。该项目将通过两种并行和互补的方法实现这一目标。首先，我们将通过一种高效的转染方法将活动依赖的荧光结构体传递到感觉神经中，根据它们的表型选择性地标记这些神经元。对这些神经元进行基于表型的富集和收集，然后进行比较表达谱分析，得到表型阳性神经元中富集的差异表达基因列表，这些基因将进行二次测试。第二种方法将应用实验室开发的新型全基因组筛选技术。这利用异源表达系统通过随机，基因激活诱变来鉴定这种新的机械激活蛋白。这两种平行的方法将相互补充，并对机械转导的分子基础产生重要的新见解。

How do mammals detect the harmful nature of blunt force, or sharp objects?How does the body defensively increase sensitivity of injured or inflamed tissues, making otherwise innocuous movements and touch feel painful? And how, as an inevitable consequence of aging, does the latter process go wrong over time, creating burdensome chronic pain? To answer these questions, we must look at sensory nerves. These cells supply our organs (including the skin) with the ability to detect information about the physical and chemical nature of our environments. They do so via protein molecules called receptors, tuned to detect specific types of environmental cues. In the past decade we havelearned much about receptors that detect heat, cold, and light touch. Those responsible for painful touch, however, remain elusive. Uncovering and characterizing these molecules carries a high reward. Beyond furthering our knowledge of sensory physiology, these would provide potential targets for the development of new, more effective pain-relief therapies with fewer side-effects. This project will use state-of-the-art technologyin functional genomics and electrophysiology to identify and study such proteins.The overall goal of the project is the identification of novel molecular receptors within the sensory nervous system, with a particular focus on those of noxious mechanical stimuli. The project will deliver this goal by two parallel and complementary approaches. First, we will deliver activity-dependent fluorescent constructs into sensory nerves via a high-efficiency transfection method to selectively label these neurones based on their phenotype. Phenotype-based enrichment and collection of these neurons will be followed by comparative expression profiling, yielding a list of differentially expressed genes enriched in phenotypically positive neurons which will be subject to secondary testing. The second approach will apply novel genome-wide screening technology developed in the lab. This utilises heterologous expression systems for the identification of such novel mechanically activated proteins via random, gene-activating mutagenesis. These two parallel approaches will complement each other and yield significant novel insight into the molecular underpinning of mechanotransduction.