Van Dam A M, Bauer J, Man-A-Hing W K, Marquette C, Tilders F J, Berkenbosch F
Department of Pharmacology, Research Institute Neurosciences Free University, Faculty of Medicine, Amsterdam, The Netherlands.
J Neurosci Res. 1995 Feb 1;40(2):251-60. doi: 10.1002/jnr.490400214.
The cellular localization of inducible (iNOS) and constitutive (cNOS) nitric oxide synthase was studied in rats by immunocytochemical techniques involving specific iNOS and cNOS directed antibodies and by NADPH-diaphorase histochemistry. Paraformaldehyde-fixed vibratome sections of brains and cryostat sections of peripheral lymph nodes were studied of rats treated with endotoxin (2.5 micrograms/kg or 2.5 mg/kg i.v.), rats infected with rabies virus, and rats exposed to experimental allergic encephalomyelitis (EAE). Endotoxin-treated animals showed no appearance of immunoreactive iNOS (ir-iNOS) cells in the brain with the exception of a few microglial cells near the median eminence and some meningeal macrophages. In the same animals however, iNOS-immunoreactive cells were found in peripheral lymph nodes. Neurons that stain positive for cNOS and for NADPH-diaphorase could be observed in brains of control as well as of endotoxin-treated animals with a similar distribution and staining intensity. In contrast, animals that had been infected with rabies virus or subjected to EAE, showed the appearance of ir-iNOS-positive cells in several brain areas. These cells are located near blood vessels and lesion sites. The majority of these cells are GSA-I-B4 isolectin-positive and therefore are likely to represent macrophages. Our data suggest that increased production of nitric oxide may play a role in the altered brain functions in rabies-infected and EAE rats. On the contrary, increased nitric oxide production is probably not involved in the non-specific symptoms of sickness induced by endotoxin.
采用免疫细胞化学技术,利用针对诱导型(iNOS)和组成型(cNOS)一氧化氮合酶的特异性抗体,并结合NADPH - 黄递酶组织化学方法,对大鼠体内诱导型和组成型一氧化氮合酶的细胞定位进行了研究。研究了用内毒素(2.5微克/千克或2.5毫克/千克静脉注射)处理的大鼠、感染狂犬病病毒的大鼠以及暴露于实验性变态反应性脑脊髓炎(EAE)的大鼠的脑甲醛固定振动切片和外周淋巴结冰冻切片。除了在正中隆起附近的一些小胶质细胞和一些脑膜巨噬细胞外,内毒素处理的动物脑内未出现免疫反应性iNOS(ir - iNOS)细胞。然而,在同一动物的外周淋巴结中发现了iNOS免疫反应性细胞。在对照动物和内毒素处理动物的脑中,均可观察到cNOS和NADPH - 黄递酶染色阳性的神经元,其分布和染色强度相似。相比之下,感染狂犬病病毒或患EAE的动物在几个脑区出现了ir - iNOS阳性细胞。这些细胞位于血管和病变部位附近。这些细胞大多数为GSA - I - B4异凝集素阳性,因此可能代表巨噬细胞。我们的数据表明,一氧化氮产生增加可能在狂犬病感染大鼠和EAE大鼠脑功能改变中起作用。相反,一氧化氮产生增加可能与内毒素诱导的非特异性疾病症状无关。