In vivo injection of α-bungarotoxin to improve the efficiency of motor endplate labeling.
In vivo injection of α-bungarotoxin to improve the efficiency of motor endplate labeling.
复制标题
体内注射α-银环蛇毒素以提高运动终板标记的效率。
DOI：
10.1002/brb3.468
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发表时间：
2016-06
影响因子：
3.1
通讯作者：
Jiang B
中科院分区：
文献类型：
--
作者：
Chen W;Yu T;Chen B;Qi Y;Zhang P;Zhu D;Yin X;Jiang B
运动终板由运动神经元终端和肌纤维组成，分布于骨骼肌中，引起肌肉收缩。然而，传统的运动终板染色方法仅限于局部骨骼肌的观察。由于严格的孵育条件，该过程非常耗时，并且通常提供令人不满意的结果。我们探索了一种通过体内注射荧光α‐班加罗毒素快速标记运动终板的新方法。将52只小鼠随机分为两组，实验组（n = 50）和对照组（n = 2）。实验组经尾静脉注射α‐班加罗毒素。注射剂量分别为0.1、0.2、0.3、0.4、0.5 μg/g。实验小鼠分为5个亚组，每组10只。对照组只注射生理盐水200 μL。取双侧腓肠肌进行显微镜分析和光学清除，寻找特异性荧光信号。0.3 μg/g α‐邦加毒素，偶联时间1 h，可在平面上显示运动终板的数量和结构。与传统方法相比，该方法快速、方便、省时。结合光学清除技术，还可以看到空间分布，有助于更好地了解骨骼肌运动终板位置的立体视图。实验证明，体内注射α‐班加罗毒素对骨骼肌运动终板的研究是有效的。
Motor endplates are composed of a motor neuron terminal and muscle fiber and are distributed in skeletal muscle, causing muscle contraction. However, traditional motor endplate staining methods are limited to the observation of partial skeletal muscle. The procedure was time‐consuming due to strict incubation conditions, and usually provided unsatisfactory results. We explored a novel method to label motor endplate rapidly by in vivo injection of fluorescent α‐bungarotoxin. Fifty‐two mice were randomly divided into two groups, an experiment group (n = 50), and a contrast group (n = 2). In experiment group, α‐bungarotoxin was injected via the caudal vein. The injection dosages were designated as 0.1, 0.2, 0.3, 0.4, and 0.5 μg/g. The experimental mice were divided into five subgroups of ten mice per group. The contrast group was only injected with 200 μL normal saline solution. Bilateral gastrocnemius were acquired for microscope analysis and optical clearing to seek specific fluorescent signal. A dose of 0.3 μg/g of α‐bungarotoxin with 1 h conjugation time could display the number and structure of motor endplate in plane view. Compared with the traditional procedure, this method was rapid, convenient, and time‐saving. Combined with the optical clearing technique, spatial distribution could also be seen, helping to better understand the stereoscopic view of motor endplate position in skeletal muscle. In vivo injection of α‐bungarotoxin proved effective for studying motor endplate in skeletal muscle.