Ruan R S, Leong S K, Yeoh K H
Department of Otolaryngology, National University Hospital, Singapore.
J Hirnforsch. 1997;38(4):433-41.
The distributions of nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) and nitric oxide synthase (NOS) in mammalian cochlea were studied at light and electron microscope levels by NADPH-d histochemistry and brain NOS (bNOS) immunohistochemistry. The cochleae from 15 albino guinea pigs were perilymphatically fixed with 2% periodate-lysine-paraformaldehyde, decalcified in 10% EDTA and processed for light and electron microscopy after NADPH-d or NOS staining in frozen and vibratome sections respectively. One human cochlea was available for light microscope examination of NADPH-d or bNOS stained sections. Light microscope results revealed that type I neurons and nerve fibers of the spiral ganglion cells were labeled by bNOS immunohistochemistry as well as NADPH-d histochemistry in both guinea pig and human cochleae. At subcellular level, NADPH-d reaction product was localized in the mitochondria of the neuronal cytoplasm and axoplasm and in the cytoplasm of the vascular endothelium. The immunoreaction products of bNOS were evenly distributed in the neuronal cytoplasm and axoplasm. Myelinated and unmyelinated fibers in the intraganglionic spiral bundle and the inner spiral and inner radial fibers below the inner hair cells were labeled for bNOS. The nerve endings below the outer hair cells were not stained. NOS immunoreaction product was also found in the outer hair cells, Schwann cells of myelinated nerve fibers, Deiter's cells, pillar cells and the tympanic lamina cells. No difference was found in the staining pattern of both NADPH-d and NOS reaction products between human and guinea pig cochleae at the light microscope level. The results suggest that NO plays an important role in the maintenance of auditory function in the mammal.
采用还原型辅酶Ⅱ黄递酶(NADPH-d)组织化学和脑型一氧化氮合酶(bNOS)免疫组织化学方法,在光镜和电镜水平上研究了烟酰胺腺嘌呤二核苷酸磷酸二酯酶(NADPH-d)和一氧化氮合酶(NOS)在哺乳动物耳蜗中的分布。用2%高碘酸盐-赖氨酸-多聚甲醛经外淋巴固定15只白化豚鼠的耳蜗,10%乙二胺四乙酸(EDTA)脱钙,分别在冰冻切片和振动切片上进行NADPH-d或NOS染色后,进行光镜和电镜处理。有1个人类耳蜗用于光镜观察NADPH-d或bNOS染色切片。光镜结果显示,在豚鼠和人类耳蜗中,螺旋神经节细胞的Ⅰ型神经元和神经纤维均被bNOS免疫组织化学以及NADPH-d组织化学标记。在亚细胞水平,NADPH-d反应产物定位于神经元细胞质和轴质的线粒体以及血管内皮细胞的细胞质中。bNOS的免疫反应产物均匀分布于神经元细胞质和轴质中。螺旋神经节内束以及内毛细胞下方的内螺旋纤维和内放射状纤维中的有髓和无髓纤维被标记为bNOS。外毛细胞下方的神经末梢未被染色。在有髓神经纤维的施万细胞、Dieters细胞、柱细胞和鼓膜层细胞中也发现了NOS免疫反应产物。在光镜水平上,人类和豚鼠耳蜗的NADPH-d和NOS反应产物的染色模式没有差异。结果表明,NO在哺乳动物听觉功能的维持中起重要作用。