Microfluidic platform for electrophysiological recordings from host-stage hookworm and Ascaris suum larvae: A new tool for anthelmintic research.
Microfluidic platform for electrophysiological recordings from host-stage hookworm and Ascaris suum larvae: A new tool for anthelmintic research.
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用于宿主阶段钩虫和猪蛔虫幼虫电生理记录的微流体平台：驱虫研究的新工具。
DOI：
10.1016/j.ijpddr.2016.08.001
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
Hawdon,JohnM
中科院分区：
文献类型：
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作者：
Weeks,JanisC;Roberts,WilliamM;Robinson,KristinJ;Keaney,Melissa;Vermeire,JonJ;UrbanJr,JosephF;Lockery,ShawnR;Hawdon,JohnM
筛选具有抗蠕虫活性的候选化合物和天然产物对于发现抗人和动物寄生虫的新药具有重要意义。我们先前验证了在秀丽隐杆线虫微流体装置（“芯片”），记录非侵入性的微小的电生理信号所产生的有节奏的收缩（泵）的蠕虫的咽。每个芯片同时记录多个蠕虫的这些咽电图（EPG），提供肌肉和神经活动的中等通量读数，这对靶向神经递质受体和离子通道的化合物特别有用。微流控技术改变了线虫的研究，本研究的目的是验证钩虫和猪蛔虫宿主期幼虫在微流控EPG平台上的活性，体外激活的钩虫和犬钩虫感染性L3 s（iL 3s）在测试条件下通常产生不稳定的EPG活性。相比之下，从仓鼠中回收的锡兰曲霉L4表现出稳健、持续的EPG活性，由三种波形组成：（1）在其他线虫中观察到的常规泵;（2）快速电压偏转，与食管的不规则收缩和食管-肠瓣的打开（称为“扑动”）相关;和（3）混合波形，我们将其归类为泵。对于数据分析，将泵和颤振合并，并将其称为EPG“事件”。EPG波形识别和分析使用定制设计的软件进行半自动化。神经调节剂5-羟色胺（5-HT）在0.5 mM的最佳浓度下增加了锡兰曲霉L4 s的EPG事件频率。驱虫药伊维菌素（IVM）以浓度依赖性方式抑制EPG活性。从猪肺中回收的猪L3 s的EPG在1 mM 5 HT中表现出强的咽部泵送，其被IVM抑制。这些实验验证了利用微流控EPG平台对锡兰曲霉L4和猪曲霉L3的使用，为筛选驱虫剂候选物或研究寄生线虫摄食行为提供了新的工具。
The screening of candidate compounds and natural products for anthelmintic activity is important for discovering new drugs against human and animal parasites. We previously validated inCaenorhabditis elegansa microfluidic device (‘chip’) that records non-invasively the tiny electrophysiological signals generated by rhythmic contraction (pumping) of the worm's pharynx. These electropharyngeograms (EPGs) are recorded simultaneously from multiple worms per chip, providing a medium-throughput readout of muscular and neural activity that is especially useful for compounds targeting neurotransmitter receptors and ion channels. Microfluidic technologies have transformedC. elegansresearch and the goal of the current study was to validate hookworm andAscaris suumhost-stage larvae in the microfluidic EPG platform.Ancylostoma ceylanicumandA. caninuminfective L3s (iL3s) that had been activatedin vitrogenerally produced erratic EPG activity under the conditions tested. In contrast,A. ceylanicumL4s recovered from hamsters exhibited robust, sustained EPG activity, consisting of three waveforms: (1) conventional pumps as seen in other nematodes; (2) rapid voltage deflections, associated with irregular contractions of the esophagus and openings of the esophogeal-intestinal valve (termed a ‘flutter’); and (3) hybrid waveforms, which we classified as pumps. For data analysis, pumps and flutters were combined and termed EPG ‘events.’ EPG waveform identification and analysis were performed semi-automatically using custom-designed software. The neuromodulator serotonin (5-hydroxytryptamine; 5HT) increased EPG event frequency inA. ceylanicumL4s at an optimal concentration of 0.5 mM. The anthelmintic drug ivermectin (IVM) inhibited EPG activity in a concentration-dependent manner. EPGs fromA. suumL3s recovered from pig lungs exhibited robust pharyngeal pumping in 1 mM 5HT, which was inhibited by IVM. These experiments validate the use ofA. ceylanicumL4s andA. suumL3s with the microfluidic EPG platform, providing a new tool for screening anthelmintic candidates or investigating parasitic nematode feeding behavior.