Herzog J, Kümmel H
Department of Physiology, Christian-Albrechts-Universität zu Kiel, Olshausenstrasse 40, D-24098, Kiel, Germany.
J Neurosci Methods. 2000 Sep 15;101(2):149-56. doi: 10.1016/s0165-0270(00)00261-2.
The transsynaptic transport of WGA-HRP is a powerful tool for the identification of last order interneurones in the spinal cord, but differentiation of primarily labelled motoneurones from transsynaptically labelled interneurones can be difficult. We therefore combined the transsynaptic labelling of interneurones with WGA-HRP and the labelling of motoneurones with fluorescent dyes (fluorogold, red beads, fast blue, DAPI) by injecting a mixture of the tracers into the nervi mediani of rats. The influence of different fixatives on the simultaneous preservation of WGA-HRP and fluorescence is evaluated. It is shown that, of several perfusion protocols, only perfusion with paraformaldehyde in addition to 1.4% lysine and 0.23% periodate allowed simultaneous visualisation of transsynaptic WGA-HRP and fluorescent dyes.
WGA-HRP的跨突触运输是鉴定脊髓中最后一级中间神经元的有力工具，但区分主要标记的运动神经元和跨突触标记的中间神经元可能会很困难。因此，我们通过将示踪剂混合物注射到大鼠正中神经中，将中间神经元的跨突触标记与WGA-HRP以及用荧光染料（荧光金、红色珠子、快蓝、DAPI）对运动神经元进行标记相结合。评估了不同固定剂对同时保存WGA-HRP和荧光的影响。结果表明，在几种灌注方案中，只有除1.4%赖氨酸和0.23%高碘酸盐外再用多聚甲醛灌注，才能同时观察到跨突触WGA-HRP和荧光染料。
