Endothelial nitric oxide synthase and caveolin-1 are co-localized in sinusoidal endothelial fenestrae
Endothelial nitric oxide synthase and caveolin-1 are co-localized in sinusoidal endothelial fenestrae
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DOI：
10.1034/j.1600-0676.2001.021003198.x
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发表时间：
2001-06-01
期刊：
影响因子：
--
通讯作者：
Ishii, H
中科院分区：
文献类型：
--
作者：
Yokomori, H;Oda, M;Ishii, H
背景/目标：一氧化氮是由钙调蛋白依赖性一氧化氮合酶 (NOS) 家族在多种哺乳动物组织中合成的。 Caveolin 是小窝中的主要结构蛋白，与内皮 NOS 相互作用，导致 Ca(++)-钙调蛋白调节的可逆过程中的酶抑制。本研究的目的是通过电子免疫金法阐明肝窦内皮细胞中 eNOS 和 Caveolin-1 的超微结构定位。方法：使用雄性Wistar大鼠。研究了通过胶原酶输注从大鼠肝脏分离的肝组织和肝窦内皮细胞。对于免疫组织化学，肝脏标本与抗 eNOS 或抗 Caveolin-1 抗体发生反应。 eNOS 或 Caveolin-1 的超微结构定位通过电子显微镜使用免疫金后嵌入方法进行鉴定。结果：使用肝组织肝窦衬里细胞、门静脉和肝动脉中的局部内皮 NOS 进行免疫组织化学研究；和caveolin-1存在于正弦衬里细胞、胆小管、门静脉和肝动脉中。在肝组织和分离的肝窦内皮细胞的肝窦内皮窗孔的质膜上显示了表明 eNOS 和 Caveolin-1 存在的免疫金颗粒。结论：内皮 NOS 和小窝蛋白共定位于窦状内皮窗孔，表明两者的相互作用可能调节 NO 合成的细胞调节。
Background/Aims: Nitric oxide is synthesized in diverse mammalian tissues by a family of calmodulin-dependent nitric oxide synthases (NOS). Caveolin, the principal structural protein in caveolae, interacts with endothelial NOS leading to enzyme inhibition in a reversible process modulated by Ca(++)-calmodulin. The aim of the present study was to clarify the ultrastructural localization of eNOS and caveolin-1 in hepatic sinusoidal endothelium by an electron immunogold method. Methods: Male Wistar rats were used. Liver tissues and hepatic sinusoidal endothelial cells isolated from rat livers by collagenase infusion were studied. For immunohistochemistry, liver specimens were reacted with anti-eNOS or anti-caveolin-1 antibody. The ultrastructural localization of eNOS or caveolin-1 was identified by electron microscopy using an immunogold postembedding method. Results: Immunohistochemical studies using liver tissues localized endothelial NOS in hepatic sinusoidal lining cells, portal veins and hepatic arteries; and caveolin-1 in sinusoidal lining cells, bile canaliculi, portal vein and hepatic arteries. Immunogold particles indicating the presence of eNOS and caveolin-1 were demonstrated on the plasma membrane of sinusoidal endothelial fenestrae in liver tissue and also in isolated sinusoidal endothelial cells. Conclusion: Endothelial NOS and caveolin are co-localized on sinusoidal endothelial fenestrae, suggesting that interaction of the two may modulate cellular regulation of NO synthesis.