Chen Z, Zandonatti M, Jakubowski D, Fox H S
Department of Neuropharmacology, Scripps Research Institute, La Jolla, California 92037, USA.
Lab Invest. 1998 Mar;78(3):353-63.
Brain capillary endothelial cells compose the blood-brain barrier, which has a crucial role in maintaining the normal extracellular environment of the central nervous system. We have developed a method to isolate endothelial cells from mouse brain and maintain them in a relatively pure primary culture. Using a subtractive hybridization technique, a novel cDNA, termed MBEC1 (mouse brain endothelial cell 1), has been isolated from the cultured mouse brain capillary endothelial cells. MBEC1 is a 1435-bp cDNA predicted to encode a protein of 218 amino acids. The predicted protein is similar to those of the newly characterized Clostridium perfringens enterotoxin receptors and is the mouse homolog of a recently described human cDNA clone, which is hemizygously deleted in individuals with velo-cardio-facial syndrome and DiGeorge syndrome. MBEC1 was expressed in our cultured MBEC, in freshly isolated MBEC, in a variety of mouse organs, and in mouse embryos as early as embryonic Day 7. In situ hybridization and immunocytochemical analyses revealed the presence of the MBEC1 mRNA and its protein product in brain capillary endothelial cells, as well as in a subset of other endothelial and epithelial cells. Moreover, developmental regulation of expression of MBEC1 was present in respiratory epithelium. Our research thus provides a new molecule for further study of the function of normal and abnormal blood-brain barrier as well as of other specialized endothelia and epithelia.
脑毛细血管内皮细胞构成血脑屏障，这在维持中枢神经系统的正常细胞外环境中起着关键作用。我们已开发出一种从小鼠脑中分离内皮细胞并将其维持在相对纯的原代培养中的方法。利用消减杂交技术，从培养的小鼠脑毛细血管内皮细胞中分离出一种新的cDNA，称为MBEC1（小鼠脑内皮细胞1）。MBEC1是一个1435碱基对的cDNA，预计编码一个218个氨基酸的蛋白质。预测的蛋白质与新鉴定的产气荚膜梭菌肠毒素受体的蛋白质相似，是最近描述的人类cDNA克隆的小鼠同源物，该克隆在患有心脏颜面综合征和迪格奥尔格综合征的个体中半合子缺失。MBEC1在我们培养的MBEC、新鲜分离的MBEC、多种小鼠器官以及早在胚胎第7天的小鼠胚胎中均有表达。原位杂交和免疫细胞化学分析显示，MBEC1 mRNA及其蛋白质产物存在于脑毛细血管内皮细胞以及其他一些内皮细胞和上皮细胞中。此外，MBEC1的表达在呼吸道上皮中存在发育调控。因此，我们的研究为进一步研究正常和异常血脑屏障以及其他特殊内皮细胞和上皮细胞的功能提供了一个新分子。
