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首页> 外文期刊>Journal of Neuroscience Methods >Labeling blastomeres with a calcium indicator: a non-invasive method of visualizing neuronal activity in zebrafish.
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Labeling blastomeres with a calcium indicator: a non-invasive method of visualizing neuronal activity in zebrafish.

机译：用钙指示剂标记卵裂球：一种在斑马鱼中可视化神经元活动的非侵入性方法。

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摘要

Injections of the calcium indicator calcium green dextran (CGD) into zebrafish embryos at the 1-4 cell stages were used to monitor the activity of neurons in larval fish. Dye was pressure injected into a single cell and the fish allowed to develop until post-hatching, when they were embedded in agar and viewed under a confocal microscope. Labeled larval cells, including identifiable neuronal classes such as Rohon-Beard cells and olfactory neurons, were clearly visible with extensive labeling of the whole fish following injections at the one cell embryonic stage, and a mosaic labeling pattern following injections at the 2 or 4 cell stages. Activity of neurons in the spinal cord, as indicated by intracellular calcium concentration changes, was observed directly by monitoring fluorescence changes of individual spinal neurons and groups of spinal neurons on a confocal microscope. Fluorescence increases of between 9 and 55% in spinal neurons were seen during escape responses produced when the fish was tapped on the tail. This technique can potentially be used to monitor the activity of any neuron or group of neurons with respect to behavior non-invasively in intact living zebrafish.
机译：在1-4个细胞阶段向斑马鱼胚胎中注射钙指示剂钙绿右旋糖酐（CGD）来监测幼鱼中神经元的活性。将染料加压注射到单个细胞中，让鱼发育直至孵化后，然后将它们包埋在琼脂中并在共聚焦显微镜下观察。标记的幼虫细胞（包括可识别的神经元类，如Rohon-Beard细胞和嗅觉神经元）清晰可见，在单个细胞胚胎阶段注射后对整个鱼进行了广泛标记，在2或4细胞注射后进行了镶嵌标记阶段。通过共聚焦显微镜监测单个脊髓神经元和脊髓神经元组的荧光变化，可以直接观察到细胞内钙浓度变化所指示的脊髓中神经元的活性。在鱼被尾巴拍打时产生的逃避反应期间，脊髓神经元的荧光增加了9％至55％。相对于在完整的活斑马鱼中的非侵入性行为，该技术可以潜在地用于监视任何神经元或神经元组的活动。

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