Ladic L A, Buchan A M
Department of Physiology, University of British Columbia, 2146 Health Sciences Mall, Vancouver BC, Canada.
Brain Res. 1998 Jun 8;795(1-2):312-24. doi: 10.1016/s0006-8993(98)00299-6.
Retrograde tracing, multi-label fluorescence immunohistochemistry, confocal microscopy and three-dimensional (3-D) reconstruction techniques were combined to examine the spatial relationship of immunoreactive nerve terminals containing either calcitonin gene-related polypeptide (CGRP) or substance P (SP) to identified gastric efferent neurons in the dorsal motor nucleus of the vagus (DMV) in the brainstem of the rat. The availability of an antibody to the receptor for SP (NK-1r) permitted observation of the association between peptide and receptor. Although both SP-IR and CGRP-IR nerve fibres came in close spatial proximity to identified gastric efferent neurons, few discrete contacts between these fibres and the neuronal membrane were observed. In addition, NK-1r-IR was localized to the somatic and dendritic membranes of a subpopulation of neurons within the DMV, with the majority of receptor labelling not in close spatial proximity to SP-IR nerve fibres. The methodology described in this study permitted the simultaneous observation of the spatial relationship between neuropeptide and an identified neuron (and the corresponding receptor in the case of SP) in 3-D, which is something that cannot be achieved using conventional microscopic techniques
将逆行追踪、多标记荧光免疫组织化学、共聚焦显微镜和三维（3-D）重建技术相结合，以研究大鼠脑干迷走神经背核（DMV）中含有降钙素基因相关肽（CGRP）或P物质（SP）的免疫反应性神经末梢与已鉴定的胃传出神经元之间的空间关系。针对SP受体（NK-1r）的抗体使观察肽与受体之间的关联成为可能。尽管SP免疫反应性（SP-IR）和CGRP免疫反应性（CGRP-IR）神经纤维在空间上都与已鉴定的胃传出神经元紧密相邻，但在这些纤维与神经元膜之间几乎未观察到离散的接触。此外，NK-1r免疫反应性（NK-1r-IR）定位于DMV内一部分神经元的胞体和树突膜上，大多数受体标记与SP-IR神经纤维在空间上并不紧密相邻。本研究中描述的方法允许在三维空间中同时观察神经肽与已鉴定神经元之间的空间关系（对于SP而言，还包括相应的受体），这是使用传统显微镜技术无法实现的。
