Ramos M, Lamé M W, Segall H J, Wilson D W
Department of Veterinary Medicine: Pathology, Immunology, Microbiology, One Shields Avenue, 1044 Haring Hall, University of California, Davis, Davis, California 95616-8617, United States.
Vascul Pharmacol. 2006 Jan;44(1):50-9. doi: 10.1016/j.vph.2005.09.007. Epub 2005 Nov 3.
Polymorphic mutations in the Bone Morphogenetic Protein type II receptor (BMPrII) gene have been implicated in the development of familial primary pulmonary hypertension (PPH) however, the role BMPrII mutations play in the development of PH has not yet been elucidated. Endothelial caveolae are an important domain of hemodynamics containing eNOS, the serotonin transporter, and endothelin receptors. In this study we show by standard immunohistochemistry (IHC) that BMPrII is widely distributed in the vasculature of the rat lung, and more specifically distributed to both apical and basal membranes of the arteriolar endothelium by fluorescent IHC. We also examined compartmentalization of BMPrII in lipid fractions of plasma membranes isolated by silica based extraction from human pulmonary artery endothelial cells and rat lung endothelium. Density gradient centrifugation demonstrated BMPrII in separate caveolin-1 (cav-1) and non-cav-1 lipid rich fractions. Electron microscopy co-localized cav-1 and BMPrII in flask shaped membrane fragments. Three-dimensional fluorescence microscopy demonstrated BMPrII in discrete membrane foci, a portion of which were co-localized with cav-1, as well as in Golgi. Our findings indicate that BMPrII is located within lipid-dense fractions of pulmonary endothelial cell membranes with a portion present in caveolae suggesting potential dynamic regulatory structural relationships.
骨形态发生蛋白II型受体(BMPrII)基因的多态性突变与家族性原发性肺动脉高压(PPH)的发生有关,然而,BMPrII突变在肺动脉高压发生中的作用尚未阐明。内皮小凹是血流动力学的一个重要区域,含有内皮型一氧化氮合酶(eNOS)、5-羟色胺转运体和内皮素受体。在本研究中,我们通过标准免疫组织化学(IHC)显示,BMPrII广泛分布于大鼠肺血管中,更具体地说,通过荧光免疫组织化学显示其分布于小动脉内皮的顶端和基底膜。我们还通过基于硅胶提取法从人肺动脉内皮细胞和大鼠肺内皮中分离的质膜脂质部分检测了BMPrII的区室化。密度梯度离心显示BMPrII存在于单独的富含小窝蛋白-1(cav-1)和非cav-1的脂质部分中。电子显微镜显示cav-1和BMPrII共定位于烧瓶状膜碎片中。三维荧光显微镜显示BMPrII存在于离散的膜灶中,其中一部分与cav-1共定位,同时也存在于高尔基体中。我们的研究结果表明,BMPrII位于肺内皮细胞膜的脂质致密部分,其中一部分存在于小凹中,提示可能存在潜在的动态调节结构关系。