Health and Biosecurity, Commonwealth Scientific and Industrial Research Organization, Geelong, VIC, Australia.
Institute for Intelligent Systems Research and Innovation, Deakin University, Geelong, VIC, Australia.
Sci Rep. 2018 Jul 4;8(1):10109. doi: 10.1038/s41598-018-28357-2.
Continuous cell lines from insect larval tissues are widely used in different research domains, such as virology, insect immunity, gene expression, and bio pharmacology. Previous study showed that introduction of 20-hydroxyecdysone to Spodoptera cell line induced a neuron-like morphology with neurite extensions. Despite some results suggesting potential presence of neuro-receptors, no study so far has shown that these neuron-induced cells were functional. Here, using microelectrode arrays, we showed that the mosquito cell line, RML12, differentiated with 20-hydroxyecdysone, displays spontaneous electrophysiological activity. Results showed that these cells can be stimulated by GABAergic antagonist as well as nicotinic agonist. These results provide new evidence of neuron-like functionality of 20-hydroxyecdysone induced differentiated mosquito cell line. Finally, we used this new model to test the effects of two insecticides, temephos and permethrin. Our analysis revealed significant changes in the spiking activity after the introduction of these insecticides with prolonged effect on the neuronal activity. We believe that this differentiated mosquito neuronal cell model can be used for high-throughput screening of new pesticides on insect nervous system instead of primary neurons or in vivo studies.
昆虫幼虫组织的连续细胞系广泛应用于不同的研究领域，如病毒学、昆虫免疫、基因表达和生物药理学。先前的研究表明，20-羟基蜕皮激素的引入诱导 Spodoptera 细胞系呈现出神经元样形态，并延伸出神经突。尽管有一些结果表明可能存在神经受体，但迄今为止没有研究表明这些诱导的神经元样细胞具有功能性。在这里，我们使用微电极阵列表明，用 20-羟基蜕皮激素诱导分化的蚊子细胞系 RML12 表现出自发的电生理活性。结果表明，这些细胞可以被 GABA 能拮抗剂和烟碱激动剂刺激。这些结果为 20-羟基蜕皮激素诱导分化的蚊子细胞系具有神经元样功能提供了新的证据。最后，我们使用这个新模型来测试两种杀虫剂，涕灭威和氯菊酯的效果。我们的分析表明，在引入这些杀虫剂后，神经元的放电活动发生了显著变化，对神经元活动的影响持续时间较长。我们相信，这种分化的蚊子神经元细胞模型可以用于昆虫神经系统的新型杀虫剂的高通量筛选，而无需使用原代神经元或体内研究。
