Re-Classification of Drosophila melanogaster Trichoid and Intermediate Sensilla Using Fluorescence-Guided Single Sensillum Recording.
Re-Classification of Drosophila melanogaster Trichoid and Intermediate Sensilla Using Fluorescence-Guided Single Sensillum Recording.
复制标题
使用荧光引导的单个感觉记录对果蝇的果蝇和中间感觉进行重新分类。
DOI：
10.1371/journal.pone.0139675
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Potter CJ
中科院分区：
文献类型：
--
作者：
Lin CC;Potter CJ
果蝇嗅觉受体神经元存在于触角和上颌须的专门感觉毛中。将气味引起的反应与嗅觉神经元活动联系起来通常是通过单感觉器记录（SSR）来完成的，其中插入单个感觉毛发的电极记录该感觉器中所有神经元的神经元活动。识别记录的感器需要将神经元反应与已知的气味反应特征相匹配。要从特定的感器进行记录，或系统地筛选所有感器类型，需要重复和半随机的 SSR 实验。在这里，我们验证了一种使用 GAL4/UAS 二元表达系统来针对特定感器进行记录的方法。我们利用现有的 OrX-Gal4 系，结合最近生成的强膜靶向 GFP 报告基因，将电生理记录引导至 GFP 标记的感器。我们验证了全套试剂，可用于快速筛选所有基本感器、中间感器和毛状感器的气味响应曲线。荧光引导的 SSR 进一步揭示了两种触角毛状感器类型应重新分类为中间感器。这种方法为研究嗅觉神经元的成熟方法提供了简单实用的补充，并且可以通过添加 UAS-geneX 效应器进行扩展，以进行功能获得或功能丧失研究。
Drosophila olfactory receptor neurons are found within specialized sensory hairs on antenna and maxillary palps. The linking of odorant-induced responses to olfactory neuron activities is often accomplished via Single Sensillum Recordings (SSR), in which an electrode inserted into a single sensory hair records the neuronal activities of all the neurons housed in that sensillum. The identification of the recorded sensillum requires matching the neuronal responses with known odor-response profiles. To record from specific sensilla, or to systematically screen all sensillar types, requires repetitive and semi-random SSR experiments. Here, we validate an approach in which the GAL4/UAS binary expression system is used for targeting specific sensilla for recordings. We take advantage of available OrX-Gal4 lines, in combination with recently generated strong membrane targeted GFP reporters, to guide electrophysiological recordings to GFP-labeled sensilla. We validate a full set of reagents that can be used to rapidly screen the odor-response profiles of all basiconic, intermediate, and trichoid sensilla. Fluorescence-guided SSR further revealed that two antennal trichoid sensilla types should be re-classified as intermediate sensilla. This approach provides a simple and practical addition to a proven method for investigating olfactory neurons, and can be extended by the addition of UAS-geneX effectors for gain-of-function or loss-of-function studies.