Youssef Khaled, Archonta Daphne, Kubiseski Terrance J, Tandon Anurag, Rezai Pouya
Department of Mechanical Engineering, York University, Toronto, ON, Canada.
Department of Biology, York University, Toronto, ON, Canada.
Lab Chip. 2021 Mar 9;21(5):821-834. doi: 10.1039/d0lc00964d.
In this paper, the novel effect of electric field (EF) on adult C. elegans egg-laying in a microchannel is discovered and correlated with neural and muscular activities. The quantitative effects of worm aging and EF strength, direction, and exposure duration on egg-laying are studied phenotypically using egg-count, body length, head movement, and transient neuronal activity readouts. Electric egg-laying rate increases significantly when worms face the anode and the response is EF-dependent, i.e. stronger (6 V cm-1) and longer EF (40 s) exposure result in a shorter egg laying response duration. Worm aging significantly deteriorates the electric egg-laying behaviour with an 88% decrease in the egg-count from day-1 to day-4 post young-adult stage. Fluorescent imaging of intracellular calcium dynamics in the main parts of the egg-laying neural circuit demonstrates the involvement and sensitivity of the serotonergic hermaphrodite specific neurons (HSNs), vulva muscles, and ventral cord neurons to the EF. HSN mutation also results in a reduced rate of electric egg-laying allowing the use of this technique for cellular screening and mapping of the neural basis of electrosensation in C. elegans. This novel assay can be parallelized and performed in a high-throughput manner for drug and gene screening applications.
在本文中，发现了电场（EF）对微通道中成年秀丽隐杆线虫产卵的新效应，并将其与神经和肌肉活动相关联。使用产卵计数、体长、头部运动和瞬时神经元活动读数，从表型上研究了线虫衰老以及EF强度、方向和暴露持续时间对产卵的定量影响。当线虫面对阳极时，电产卵率显著增加，且该反应依赖于EF，即更强的（6 V cm-1）和更长时间的EF（40 s）暴露会导致产卵反应持续时间缩短。从年轻成虫阶段后的第1天到第4天，线虫衰老显著恶化电产卵行为，产卵计数下降88%。对产卵神经回路主要部分的细胞内钙动力学进行荧光成像，证明了5-羟色胺能雌雄同体特异性神经元（HSNs）、外阴肌肉和腹侧索神经元对EF的参与和敏感性。HSN突变也导致电产卵率降低，从而使该技术可用于秀丽隐杆线虫电感觉神经基础的细胞筛选和图谱绘制。这种新的检测方法可以并行化，并以高通量方式用于药物和基因筛选应用。
