Department of Evolutionary Neuroethology, Max Planck Institute for Chemical Ecology, Hans-Knoell Str. 8, Jena 07745, Germany.
J Neurosci Methods. 2011 Oct 15;201(2):385-9. doi: 10.1016/j.jneumeth.2011.08.015. Epub 2011 Aug 17.
Recent insights into insect olfactory signaling based on in vitro analyses have created an urgent need for equivalent in vivo analyses using living organisms. Here, we present a microinjection system that establishes a "virtual petri dish" within sensory structures for the application of agents to sensory neurons. Our system uses a series of pumps to inject chemical agents via air pressure into the surrounding lymph. We show using tetrodotoxin and forskolin application that robust effects on response dynamics of Drosophila melanogaster olfactory sensory neurons could be observed within 200 s, and suggest data analysis techniques to improve estimation of pharmacological effects on response kinetics. This approach provides an improved in vivo method to investigate questions in sensory neuron physiology as a complement to heterologous expression systems.
基于体外分析的昆虫嗅觉信号的最新研究进展,迫切需要使用活体生物进行等效的体内分析。在这里,我们提出了一种微注射系统,该系统在感觉结构内建立了一个“虚拟培养皿”,以便将试剂应用于感觉神经元。我们的系统使用一系列泵通过气压将化学试剂注入周围的淋巴液中。我们使用河豚毒素和 forskolin 的应用表明,可以在 200 秒内观察到黑腹果蝇嗅觉感觉神经元反应动力学的强烈影响,并提出了数据分析技术来提高对反应动力学的药物作用的估计。这种方法为研究感觉神经元生理学问题提供了一种改进的体内方法,可作为异源表达系统的补充。