Henschen A, Hökfelt T, Elde R, Fahrenkrug J, Frey P, Terenius L, Olson L
Department of Histology and Neurobiology, Karolinska Institutet, Stockholm, Sweden.
Neuroscience. 1988 Jul;26(1):193-213. doi: 10.1016/0306-4522(88)90137-6.
The present study examines the distribution of several neuropeptides, as revealed by immunohistochemistry in the isolated cord. Fetal rat spinal cord was grafted to the anterior chamber of the adult Sprague-Dawley albino rats. After intraocular maturation for 2-3 months, the amount and distribution of somatostatin, neuropeptide Y, substance P, enkephalin, vasoactive intestinal peptide, peptide histidine-isoleucine, calcitonin gene-related peptide and cholecystokinin immunoreactive terminals and cell bodies were analysed using indirect fluorescence immunohistochemistry. The visualization of immunoreactive cell bodies in the grafts was enhanced using a novel intraocular colchicine treatment. In the graft a rich network of somatostatin-positive terminals was found with a high density in well-demarcated areas reminiscent of substantia gelatinosa of the dorsal horn of normal spinal cord. A large number of small- to medium-sized somatostatin neurons was found throughout the grafts without colchicine treatment. This is in contrast to normal spinal cord, where positive neurons were difficult to visualize without colchicine and were mainly confined to the dorsal horn. Neuropeptide Y had a distribution in the grafts similar to that of somatostatin and neuropeptide Y cells were found throughout the grafts without colchicine treatment. In normal spinal cord, neuropeptide Y-positive fibers were found mainly in substantia gelatinosa with a sparse network in the ventral horn. Enkephalin-positive fibers were found throughout the grafts. The distribution of fibers resembled that of somatostatin and neuropeptide Y with distinct zones of high fiber density in well-demarcated areas, whereas the density of nerve fibers in the rest of the graft neuropil was moderate to low. The distribution of substance P was similar to that of enkephalin. After colchicine treatment, both enkephalin- and substance P-positive cell bodies were visualized. In the intact spinal cord both peptides were seen in the entire gray matter with the highest concentrations in the superficial laminae of the dorsal horn. Antisera against calcitonin gene related-peptide, revealed a sparse terminal network and many large cells, which might represent motoneurons. A sparse network of varicose cholecystokinin-immunoreactive fibers was found evenly distributed in the grafts. In normal spinal cord a dense cholecystokinin-positive network of primary sensory afferent origin was found in the dorsal horn. In the grafts cholecystokinin cell bodies were seen after colchicine treatment.(ABSTRACT TRUNCATED AT 400 WORDS)
本研究通过免疫组织化学方法，研究了几种神经肽在离体脊髓中的分布情况。将胎鼠脊髓移植到成年Sprague-Dawley白化大鼠的前房。眼内成熟2至3个月后，使用间接荧光免疫组织化学方法分析生长抑素、神经肽Y、P物质、脑啡肽、血管活性肠肽、肽组氨酸异亮氨酸、降钙素基因相关肽和胆囊收缩素免疫反应性终末及细胞体的数量和分布。使用一种新型的眼内秋水仙碱处理方法增强了移植体中免疫反应性细胞体的可视化效果。在移植体中发现了丰富的生长抑素阳性终末网络，在界限清晰的区域密度较高，让人联想到正常脊髓背角的胶状质。在未用秋水仙碱处理的整个移植体中发现了大量中小尺寸的生长抑素神经元。这与正常脊髓不同，在正常脊髓中，不用秋水仙碱很难看到阳性神经元，且主要局限于背角。神经肽Y在移植体中的分布与生长抑素相似，在未用秋水仙碱处理的整个移植体中都发现了神经肽Y细胞。在正常脊髓中，神经肽Y阳性纤维主要见于胶状质，腹角有稀疏网络。脑啡肽阳性纤维遍布整个移植体。纤维的分布与生长抑素和神经肽Y相似，在界限清晰的区域有明显的高纤维密度区，而移植体其余神经毡中的神经纤维密度为中度到低度。P物质的分布与脑啡肽相似。秋水仙碱处理后，脑啡肽和P物质阳性细胞体都能被可视化。在完整脊髓中，两种肽在整个灰质中都可见，在背角浅层浓度最高。抗降钙素基因相关肽的抗血清显示出稀疏的终末网络和许多大细胞，这些大细胞可能代表运动神经元。在移植体中发现了稀疏的曲张胆囊收缩素免疫反应性纤维网络，均匀分布。在正常脊髓中，在背角发现了密集的源自初级感觉传入的胆囊收缩素阳性网络。秋水仙碱处理后在移植体中可见胆囊收缩素细胞体。（摘要截短于400字）
