Janelia Farm Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, United States of America.
PLoS One. 2013 Aug 20;8(8):e71706. doi: 10.1371/journal.pone.0071706. eCollection 2013.
All organisms react to noxious and mechanical stimuli but we still lack a complete understanding of cellular and molecular mechanisms by which somatosensory information is transformed into appropriate motor outputs. The small number of neurons and excellent genetic tools make Drosophila larva an especially tractable model system in which to address this problem. We developed high throughput assays with which we can simultaneously expose more than 1,000 larvae per man-hour to precisely timed noxious heat, vibration, air current, or optogenetic stimuli. Using this hardware in combination with custom software we characterized larval reactions to somatosensory stimuli in far greater detail than possible previously. Each stimulus evoked a distinctive escape strategy that consisted of multiple actions. The escape strategy was context-dependent. Using our system we confirmed that the nociceptive class IV multidendritic neurons were involved in the reactions to noxious heat. Chordotonal (ch) neurons were necessary for normal modulation of head casting, crawling and hunching, in response to mechanical stimuli. Consistent with this we observed increases in calcium transients in response to vibration in ch neurons. Optogenetic activation of ch neurons was sufficient to evoke head casting and crawling. These studies significantly increase our understanding of the functional roles of larval ch neurons. More generally, our system and the detailed description of wild type reactions to somatosensory stimuli provide a basis for systematic identification of neurons and genes underlying these behaviors.
所有生物体对有害和机械刺激都会产生反应,但我们仍然缺乏对将躯体感觉信息转化为适当运动输出的细胞和分子机制的完整理解。由于果蝇幼虫神经元数量较少且遗传工具出色,因此它是解决这一问题的特别适用的模型系统。我们开发了高通量检测方法,每小时可同时对超过 1000 只幼虫进行精确定时的有害热、振动、气流或光遗传学刺激。通过使用该硬件和定制软件,我们对幼虫对躯体感觉刺激的反应进行了比以前更详细的描述。每种刺激都会引发独特的逃避策略,该策略由多个动作组成。逃避策略是依赖于环境的。使用我们的系统,我们证实伤害感受性 IV 类多树突神经元参与了对有害热的反应。触须神经元(ch)对于头部扭转、爬行和蜷缩对机械刺激的正常调节是必需的。我们观察到振动引起的 ch 神经元钙瞬变增加,这与上述结果一致。触须神经元的光遗传学激活足以引起头部扭转和爬行。这些研究大大增加了我们对幼虫触须神经元的功能作用的理解。更普遍地说,我们的系统和对躯体感觉刺激的野生型反应的详细描述为系统地识别这些行为背后的神经元和基因提供了基础。