Reiner M, Bloch W, Addicks K
Department of Anatomy I, University of Cologne, Cologne, Germany.
J Histochem Cytochem. 2001 Dec;49(12):1605-10. doi: 10.1177/002215540104901214.
Immunogold labeling on samples of isolated perfused rat hearts embedded by an innovative low-temperature LR White procedure provided detailed insight into the interaction of caveolin-1 and endothelial NOS in myocardial capillary endothelium at the subcellular level. Separately, the localization of caveolin-1 and eNOS at caveolae under steady state conditions was visualized. A double-labeling experiment supported their close co-localization. Short-term bradykinin stimulation caused a detectable dissociation of eNOS from caveolin and its redistribution to different cell compartments, whereas caveolin itself remained stationary at caveolae. Morphometric analysis revealed that more than 80% of detectable eNOS was co-localized with caveolin-1 at caveolae under control conditions. After brief stimulation for 2 min with 10(-7) M bradykinin, only 26% of the eNOS signals were associated with caveolin-1 and randomly distributed over the endothelial cells. After stimulation, eNOS was found at the plasmalemmal and intracellular membranes, freely in the cytoplasm, and at outer mitochondrial membranes.
采用创新的低温LR White程序包埋分离的灌注大鼠心脏样本，进行免疫金标记，在亚细胞水平上详细深入地了解了小窝蛋白-1与心肌毛细血管内皮细胞中内皮型一氧化氮合酶的相互作用。另外，还观察到了稳态条件下小窝蛋白-1和内皮型一氧化氮合酶在小窝中的定位。一项双重标记实验证实了它们紧密的共定位。短期缓激肽刺激导致内皮型一氧化氮合酶与小窝蛋白可检测到的解离，并使其重新分布到不同的细胞区室，而小窝蛋白本身则保持固定在小窝处。形态计量分析显示，在对照条件下，超过80%可检测到的内皮型一氧化氮合酶与小窝蛋白-1在小窝中共定位。用10(-7) M缓激肽短暂刺激2分钟后，只有26%的内皮型一氧化氮合酶信号与小窝蛋白-1相关，并随机分布在内皮细胞上。刺激后，在内质膜和细胞内膜、细胞质中自由以及线粒体外膜上均发现了内皮型一氧化氮合酶。
