Bööj S, Goldstein M, Fischer-Colbrie R, Dahlström A
Institute of Neurobiology, University of Göteborg, Sweden.
Neuroscience. 1989;30(2):479-501. doi: 10.1016/0306-4522(89)90267-4.
The presence and intra-axonal transport of calcitonin gene-related peptide and chromogranin A were investigated in motor neurons belonging to the rat sciatic nerve. Their co-localization with markers of cholinergic organelles (SV2, p38, and synapsin I) was also investigated, using immunofluorescence techniques, including double labelling experiments. It was found that motor perikarya in the lumbar spinal cord contained calcitonin gene-related peptide-like immunoreactivity and chromogranin A-like immunoreactivity, and probably also caligulin-like immunoreactivity, located in the Nissl substance of the cytoplasm. Also, some SV2 (detected by the monoclonal antibody 10H) was present in some motor neuron perikarya, but most often these were devoid of SV2 and p38, as well as of synapsin I-like immunoreactivity. These three antigens were, on the other hand, concentrated in nerve terminals in the entire gray substance of the spinal cord. In the ventral root, after crushing, calcitonin gene-related peptide, chromogranin A, synapsin I, SV2, p38 and caligulin-like immunoreactivity accumulated in thick and medium-sized axons proximal to the crush, while only antisera against SV2 and p38 labelled accumulated material distal to the crush. In the sciatic nerve, the same essential picture was observed as in the ventral root, but here two other nervous components were also present in the normal sciatic nerve, i.e. peripheral branches of the sensory system and axons of the sympathetic system. By various denervation procedures, it was demonstrated that most calcitonin gene-related peptide-like immunoreactivity and almost all chromogranin A-like immunoreactivity, accumulating in thick axons proximally, emanated from the ventral root. Thin and medium-sized axons originated from the sensory and sympathetic systems and contributed to accumulations both proximally and distally to the crush. Synapsin I-like immunoreactive material accumulated only proximal to the crush, while SV2 and p38-like material accumulated bidirectionally in axons of all sizes. In motor endplates of the rat diaphragm and gastrocnemic muscle, no calcitonin gene-related peptide-like material was observed. However, some chromogranin A-like immunoreactivity was present, in addition to large amounts of synapsin I-like, p38-like and SV2-like material, which had a finely granular appearance and was concentrated near the presynaptic membrane of the nerve terminal endfeet, where synaptic vesicles are known to be located.
研究了大鼠坐骨神经运动神经元中降钙素基因相关肽和嗜铬粒蛋白A的存在及轴突内运输情况。还利用免疫荧光技术，包括双重标记实验，研究了它们与胆碱能细胞器标记物（SV2、p38和突触素I）的共定位。结果发现，腰段脊髓运动神经元胞体含有降钙素基因相关肽样免疫反应性和嗜铬粒蛋白A样免疫反应性，可能还含有钙调蛋白样免疫反应性，位于细胞质的尼氏体中。此外，一些运动神经元胞体中存在一些SV2（由单克隆抗体10H检测到），但这些胞体大多缺乏SV2和p38以及突触素I样免疫反应性。另一方面，这三种抗原集中在脊髓全层灰质的神经末梢中。在腹根，挤压后，降钙素基因相关肽、嗜铬粒蛋白A、突触素I、SV2、p38和钙调蛋白样免疫反应性在挤压近端的粗、中等大小轴突中积累，而只有抗SV2和p38的抗血清标记了挤压远端的积累物质。在坐骨神经中，观察到与腹根相同的基本情况，但正常坐骨神经中还存在另外两种神经成分，即感觉系统的外周分支和交感神经系统的轴突。通过各种去神经支配程序表明，在近端粗轴突中积累的大部分降钙素基因相关肽样免疫反应性和几乎所有嗜铬粒蛋白A样免疫反应性都来自腹根。细和中等大小的轴突起源于感觉和交感神经系统，在挤压近端和远端都有积累。突触素I样免疫反应性物质仅在挤压近端积累，而SV2和p38样物质在各种大小的轴突中双向积累。在大鼠膈肌和腓肠肌的运动终板中，未观察到降钙素基因相关肽样物质。然而，除了大量突触素I样、p38样和SV2样物质外，还存在一些嗜铬粒蛋白A样免疫反应性，这些物质呈细颗粒状外观，集中在神经末梢终足的突触前膜附近，已知突触小泡位于此处。
