Brugia malayi is a human filarial nematode parasite that causes lymphatic filariasis or ‘elephantiasis’ a disfiguring neglected tropical disease. This parasite is a more tractable nematode parasite for the experimental study of anthelmintic drugs and has been studied with patch-clamp and RNAi techniques. Unlike in C. elegans however, calcium signaling in B. malayi or other nematode parasites has not been achieved, limiting the studies of the mode of action of anthelmintic drugs. We describe here the development of calcium imaging methods that allow us to characterize changes in cellular calcium in the muscles of B. malayi. This is a powerful technique that can help in elucidating the mode of action of selected anthelmintics. We developed two approaches that allow the recording of calcium signals in the muscles of adult B. malayi: (a) soaking the muscles with Fluo-3AM, promoting large-scale imaging of multiple cells simultaneously and, (b) direct insertion of Fluo-3 using microinjection, providing the possibility of performing dual calcium and electrophysiological recordings. Here, we describe the techniques used to optimize dye entry into the muscle cells and demonstrate that detectable increases in Fluo-3 fluorescence to elevated calcium concentrations can be achieved in B. malayi using both techniques.

中文翻译：

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适应技术在成年 Brugia malayi 的肌肉中进行钙成像。

马来丝虫是一种人类丝虫线虫寄生虫，可引起淋巴丝虫病或“象皮病”，这是一种被忽视的毁容热带疾病。这种寄生虫是一种更易于处理的线虫寄生虫，用于驱虫药物的实验研究，并已使用膜片钳和 RNAi 技术进行了研究。然而，与秀丽隐杆线虫不同的是，尚未在马来亚拟南芥或其他线虫寄生虫中实现钙信号传导，这限制了对驱虫药物作用方式的研究。我们在这里描述了钙成像方法的发展，这些方法使我们能够表征马来亚人肌肉中细胞钙的变化. 这是一种强大的技术，可以帮助阐明所选驱虫药的作用模式。我们开发了两种方法来记录成年马来人肌肉中的钙信号：(a) 用 Fluo-3AM 浸泡肌肉，同时促进多个细胞的大规模成像，(b) 直接插入 Fluo- 3 使用显微注射，提供了进行双钙和电生理记录的可能性。在这里，我们描述了用于优化染料进入肌肉细胞的技术，并证明使用这两种技术可以在B. malayi 中实现 Fluo-3 荧光到钙浓度升高的可检测增加。