McClanahan Patrick D, Xu Joyce H, Fang-Yen Christopher
Department of Bioengineering, University of Pennsylvania, Philadelphia, PA, USA.
Integr Biol (Camb). 2017 Oct 16;9(10):800-809. doi: 10.1039/c7ib00120g.
The roundworm Caenorhabditis elegans is an important model system for understanding the genetics and physiology of touch. Classical assays for C. elegans touch, which involve manually touching the animal with a probe and observing its response, are limited by their low throughput and qualitative nature. We developed a microfluidic device in which several dozen animals are subject to spatially localized mechanical stimuli with variable amplitude. The device contains 64 sinusoidal channels through which worms crawl, and hydraulic valves that deliver touch stimuli to the worms. We used this assay to characterize the behavioral responses to gentle touch stimuli and the less well studied harsh (nociceptive) touch stimuli. First, we measured the relative response thresholds of gentle and harsh touch. Next, we quantified differences in the receptive fields between wild type worms and a mutant with non-functioning posterior touch receptor neurons. We showed that under gentle touch the receptive field of the anterior touch receptor neurons extends into the posterior half of the body. Finally, we found that the behavioral response to gentle touch does not depend on the locomotion of the animal immediately prior to the stimulus, but does depend on the location of the previous touch. Responses to harsh touch, on the other hand, did not depend on either previous velocity or stimulus location. Differences in gentle and harsh touch response characteristics may reflect the different innervation of the respective mechanosensory cells. Our assay will facilitate studies of mechanosensation, sensory adaptation, and nociception.
蛔虫秀丽隐杆线虫是理解触觉遗传学和生理学的重要模型系统。传统的秀丽隐杆线虫触觉检测方法是用探针手动触摸动物并观察其反应,这种方法受限于低通量和定性性质。我们开发了一种微流控装置,其中几十只动物会受到可变幅度的空间局部机械刺激。该装置包含64个蠕虫爬行通过的正弦通道,以及向蠕虫传递触觉刺激的液压阀。我们使用这种检测方法来表征对轻柔触觉刺激和研究较少的强烈(伤害性)触觉刺激的行为反应。首先,我们测量了轻柔触觉和强烈触觉的相对反应阈值。其次,我们量化了野生型蠕虫和后触觉受体神经元功能缺失突变体之间感受野的差异。我们发现,在轻柔触觉下,前触觉受体神经元的感受野延伸到身体后半部。最后,我们发现对轻柔触觉的行为反应不取决于刺激前动物的运动,但确实取决于先前触摸的位置。另一方面,对强烈触觉的反应既不取决于先前的速度也不取决于刺激位置。轻柔触觉和强烈触觉反应特征的差异可能反映了各自机械感觉细胞的不同神经支配。我们的检测方法将有助于机械感觉、感觉适应和伤害感受的研究。