Whole-mount procedures for simultaneous visualization of nerves, neurons, cartilage and bone
Whole-mount procedures for simultaneous visualization of nerves, neurons, cartilage and bone
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DOI：
10.1016/s1385-299x(99)00007-0
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发表时间：
1999-07-01
期刊：
影响因子：
--
通讯作者：
Fritzsch, B
中科院分区：
文献类型：
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作者：
Rosa-Molinar, E;Proskocil, BJ;Fritzsch, B
本文描述了两种选择性标记和/或染色传入和传出纤维及其细胞体并同时染色骨和软骨的技术。一种技术使用抗乙酰化醇微管蛋白免疫组化来追踪支配轴向肌肉组织的腹侧脊神经的路线、外周分支和起源，另一种技术使用选择性应用的3 kDa生物素右旋糖酐胺的顺行和逆行运输来识别特定的传入和传出投射及其细胞体。这两种方法都可以与酶清除和染色程序相结合，以同时显示全载制剂中的骨（茜素红S）和软骨（阿利新蓝）。有髓神经纤维和无髓神经纤维用3,3'二氨基联苯胺四盐酸盐（DAB）反应产物染成深棕色，而软骨染成蓝色（阿利新蓝），骨染成红色（茜素红S）。这些程序的结合提供了对传入和传出投射及其细胞体以及轴向和尾向骨骼系统的三维地形的同时理解，这是理解脊髓相对于轴向和尾向骨骼的室间关系的关键。(C) 1999 Elsevier Science B.V.版权所有
Herein are described two techniques to selectively label and/or stain afferent and efferent fibers and their cell bodies and simultaneously stain bone and cartilage. One technique uses anti-acetylated ol-tubulin immunohistochemistry to follow the course, peripheral branching, and origin of the ventral spinal nerve innervating the axial musculature and a second uses anterograde and retrograde transport of selectively applied 3 kDa biotin dextran amines to identify specific afferent and efferent projections and their cell bodies. Both procedures can be combined with an enzyme clearing and staining procedure for the simultaneous visualization of bone (alizarin red S) and cartilage (alcian blue) in whole-mount preparations. Myelinated and unmyelinated nerve fibers are stained dark brown with the 3,3' diaminobenzidine tetrahydrochloride (DAB) reaction product, whereas cartilage is stained blue (alcian blue) and bone is red (alizarin red S). The combination of these procedures provides a simultaneous three-dimensional understanding of the topography of afferent and efferent projections and their cell bodies and the axial and appendicular skeletal system, which is key to understanding the compartmental relationships of the spinal cord with respect to the axial and appendicular skeleton. (C) 1999 Elsevier Science B.V. All rights reserved.