Kitchener P D, Bourreau J P, Diamond J
Dept. of Anatomical Sciences, University of Queensland, Brisbane, Australia.
Neuroscience. 1993 Apr;53(3):613-24. doi: 10.1016/0306-4522(93)90610-r.
In addition to labelling endothelium, some ependymal cells (including tanycytes), and a subpopulation of neurons, nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase histochemistry of stab lesion sites in the neocortex revealed a large population of cells concentrated within several hundred micrometers of the lesion site. To determine the identity of these cells, NADPH-diaphorase reactivity was compared to binding with either the I-B4 isolectin from Bandeiraea simplicifolia (which has previously been shown to identify endothelial cells and activated mononuclear phagocytes), or a monoclonal antibody (OX-42) that recognizes activated mononuclear phagocytes. Many I-B4 lectin-labelled cells were also NADPH-diaphorase reactive, and other I-B4 lectin-labelled cells were also OX-42 immunoreactive, but co-existence of OX-42 immunoreactivity and NADPH-diaphorase reactivity was not observed. Only a small minority of NADPH-diaphorase-reactive cells did not exhibit I-B4 lectin binding. In contrast to the simple somatic morphology of the majority of NADPH-diaphorase-reactive cells, the I-B4 lectin-negative cells had a ramified appearance, and while readily observed at two days postlesion, they were only rarely seen at three days postlesion. Primary cultures of bovine aortic endothelial cells also exhibited NADPH-diaphorase reactivity which occupied most of the cytoplasm in a filamentous web pattern. Endothelial cells possess a constitutive form of nitric oxide synthase which, as demonstrated in NADPH-diaphorase-reactive neurons, may be the basis of their NADPH-diaphorase reactivity. These findings indicate that NADPH-diaphorase-reactive cells observed at lesion sites are probably angiogenic endothelial cells not associated with extant blood vessels. Thus, NADPH-diaphorase histochemistry offers an effective method of visualizing neovascularization in the brain and other tissues.
除了标记内皮细胞、一些室管膜细胞（包括伸展细胞）和一部分神经元外，用烟酰胺腺嘌呤二核苷酸磷酸（NADPH）-黄递酶组织化学方法对新皮质刺伤损伤部位进行研究，结果显示大量细胞集中在损伤部位数百微米范围内。为了确定这些细胞的身份，将NADPH-黄递酶反应性与来自单叶豆的I-B4异凝集素（先前已证明可识别内皮细胞和活化的单核吞噬细胞）或识别活化单核吞噬细胞的单克隆抗体（OX-42）的结合情况进行了比较。许多I-B4凝集素标记的细胞也具有NADPH-黄递酶反应性，其他I-B4凝集素标记的细胞也具有OX-42免疫反应性，但未观察到OX-42免疫反应性和NADPH-黄递酶反应性并存的情况。只有极少数NADPH-黄递酶反应性细胞不表现出I-B4凝集素结合。与大多数NADPH-黄递酶反应性细胞简单的体细胞形态不同，I-B4凝集素阴性细胞呈分支状外观，虽然在损伤后两天很容易观察到，但在损伤后三天很少见到。牛主动脉内皮细胞的原代培养物也表现出NADPH-黄递酶反应性，其以丝状网络模式占据大部分细胞质。内皮细胞具有组成型一氧化氮合酶，正如在NADPH-黄递酶反应性神经元中所证明的那样，这可能是其NADPH-黄递酶反应性的基础。这些发现表明，在损伤部位观察到的NADPH-黄递酶反应性细胞可能是与现存血管无关的血管生成内皮细胞。因此，NADPH-黄递酶组织化学为可视化脑和其他组织中的新生血管提供了一种有效的方法。
