Transcuticular optical imaging of stimulus-evoked neural activities in the Drosophila peripheral nervous system
Transcuticular optical imaging of stimulus-evoked neural activities in the Drosophila peripheral nervous system
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DOI：
10.1038/nprot.2010.85
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发表时间：
2010-01-01
期刊：
影响因子：
14.8
通讯作者：
Fiala, Andre
中科院分区：
文献类型：
--
作者：
Kamikouchi, Azusa;Wiek, Robert;Fiala, Andre
果蝇的神经系统被广泛用于研究神经元的信号处理，因为神经元的活动可以通过基因编码的致动器和感受器蛋白的细胞类型特异性表达来控制和监测。然而，测量成年果蝇的神经活动通常需要外科手术才能穿透坚固的、有色素的角质外骨骼。对于周围神经系统(PNSPNSPNS)来说，干扰外骨骼是至关重要的，因为感觉神经元通常位于角质层的正下方，与专门的接受刺激和传导刺激的角质层结构相关。在这篇文章中，我们描述了如何通过角质层非破坏性地探测这些神经元的活动，如果使用具有强基线荧光的遗传编码的荧光蛋白传感器。该方法用于成年触角的机械感觉神经元，但也可应用于许多其他PNSPNSPNS神经元，如位于苍蝇腿上的股骨脊索器官所示。
he nervous system of Drosophila is widely used to study neuronal signal processing because the activities of neurons can be controlled and monitored by cell type-specific expression of genetically encoded actuator and sensor proteins. Measuring neural activities in adult flies, however, usually requires surgical approaches to penetrate the firm and pigmented cuticular exoskeleton. Interfering with this exoskeleton is critical in the case of the peripheral nervous system (PNSPNSPNS), as sensory neurons are often located directly beneath the cuticle and are associated with specialized stimulus-receiving and -conducting cuticular structures. In this article, we describe how the activities of these neurons can be probed nondestructively through the cuticle if a genetically encoded fluorescent protein sensor with strong baseline fluorescence is used. The method is exemplified for mechanosensory neurons in the adult antenna but can also be applied to many other PNSPNSPNS neurons, as is shown for the femoral chordotonal organ located in the fly's leg.