Identification of the neuronal effects of ethanol on C. elegans by in vivo fluorescence imaging on a microfluidic chip
Identification of the neuronal effects of ethanol on C. elegans by in vivo fluorescence imaging on a microfluidic chip
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DOI：
10.1007/s00216-010-4148-z
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发表时间：
2011-04
影响因子：
4.3
通讯作者：
Ying Wang;Jingjing Wang;Wei Du;Xiaojun Feng;Bifeng Liu
中科院分区：
文献类型：
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作者：
Ying Wang;Jingjing Wang;Wei Du;Xiaojun Feng;Bifeng Liu
秀丽隐杆线虫（Caenorhabditis elegans，C. elegans）是用于研究行为和神经元活动之间的相关性的良好建立的模式生物。在这里，我们展示了一种基于微流体的方法，允许基于刺激的神经元分析固定的C。elegans用于鉴定乙醇对右ASE（ASER）神经元的化学感受反应的神经元效应。研制了一种用于固定C.优美的模拟通过层流的界面移动来实现。将在ASER神经元中表达钙指示剂G-CaMP的良好喂养的转基因蠕虫用于体内荧光成像。为了评估所开发的方法，我们首先研究了乙醇对ASER神经元响应单一NaCl刺激的影响。结果表明，乙醇急性抑制ASER神经元对NaCl的ON反应，而不是OFF反应。进一步研究了ASER神经元对NaCl和乙醇的适应性，结果表明乙醇干扰了神经元的适应性。该方法具有操作简单、通量高的优点。我们希望这种新方法能为研究C.优美的
Caenorhabditis elegans (C. elegans) is a well-established model organism for investigating the correlations between behavioral and neuronal activities. Here, we demonstrated a microfluidic-based method that allowed stimulation-based neuronal analysis of immobilized C. elegans for identifying the neuronal effects of ethanol on the chemosensory responses of the right ASE (ASER) neuron. A one-piece microvalve was developed for the immobilization of C. elegans. Stimulations were realized by interface shifting of laminar flows. Well-fed transgenic worms expressing the calcium indicator G-CaMP in ASER neurons were used for in vivo fluorescence imaging. To evaluate the developed method, we first studied the effects of ethanol on the ASER neurons in response to a single NaCl stimulus. Results indicated that ethanol acutely suppressed the ON responses of ASER neurons to NaCl rather than the OFF response. Further studies of the adaptation of ASER neurons in response to NaCl and in the presence of ethanol suggested that ethanol interfered with the adaptation of neurons. The developed method exhibited the advantages of ease of operation and high throughput. We expect this new method to open up a new avenue for investigating the correlations between the behavioral and neuronal activities of C. elegans.