Fritzsch B
Department of Biomedical Sciences, Creighton University, Omaha, NE 68178.
J Neurosci Methods. 1993 Oct;50(1):95-103. doi: 10.1016/0165-0270(93)90060-5.
The distances of anterograde and retrograde axonal movement per hour were examined for dextran amines of 3000, 10,000 and 40,000 molecular weights (MW) conjugated to different fluorochromes (Cascade blue, fluorescein, tetramethylrhodamine, Texas red) or to biotin. Lateral line nerves of Xenopus laevis tadpoles were used as an easily accessible test system. Only 10,000 and 3000 MW dextran amines underwent significant anterograde and retrograde movement. Dextrans of 3000 MW progressed about twice as far (2 mm/h at 22 degrees C) than 10,000 MW dextrans. Dextrans conjugated to different fluorochromes or to biotin did not show differences in their distance covered. Tracers traveled over the same distance in amphibians pre-treated with either 1 microM colchicine for 2-6 h or 10 micrograms/ml nocodazole for 5 h prior to the application to depolymerize microtubules needed for active transport. This suggests that diffusion is the major mechanism of movement for dextran amines over short distances. In the developing Xenopus retina, 3000 MW dextrans traveled from the optic disc as far away as the ora serrata within 1 h. In mouse embryos, chicken embryos and larval lampreys dextran amines progress within 1 h from cut peripheral nerves through efferent and afferent tracts. These data show that 3000 MW dextrans can be used in much the same way as 10,000 MW dextrans, but label neuronal profiles in a shorter time in a wide variety of species.
研究了与不同荧光染料(级联蓝、荧光素、四甲基罗丹明、德克萨斯红)或生物素偶联的分子量为3000、10000和40000道尔顿(MW)的葡聚糖胺每小时的顺行和逆行轴突运输距离。非洲爪蟾蝌蚪的侧线神经被用作易于获取的测试系统。只有分子量为10000和3000道尔顿的葡聚糖胺经历了显著的顺行和逆行运输。3000道尔顿的葡聚糖运输速度约为10000道尔顿葡聚糖的两倍(22℃时为2毫米/小时)。与不同荧光染料或生物素偶联的葡聚糖在运输距离上没有差异。在用1微摩尔秋水仙碱预处理2 - 6小时或用10微克/毫升诺考达唑预处理5小时以解聚主动运输所需的微管后,将示踪剂应用于两栖动物,示踪剂在相同距离内运输。这表明扩散是葡聚糖胺在短距离内运输的主要机制。在发育中的非洲爪蟾视网膜中,3000道尔顿的葡聚糖在1小时内从视盘运输到锯齿缘。在小鼠胚胎、鸡胚胎和七鳃鳗幼体中,葡聚糖胺在1小时内从切断的外周神经通过传出和传入束运输。这些数据表明,3000道尔顿的葡聚糖可以与10000道尔顿的葡聚糖以大致相同的方式使用,但在更短的时间内标记多种物种的神经元轮廓。