Larre Isabel, Contreras Ruben G, Cereijido Marcelino
Center for Research & Advanced Studies (CINVESTAV), Mexico City, México.
Methods Mol Biol. 2011;763:155-68. doi: 10.1007/978-1-61779-191-8_10.
Ouabain, a toxic of vegetal origin used for centuries to treat heart failure, has recently been demonstrated to have an endogenous counterpart, most probably ouabain itself, which behaves as a hormone. Therefore, the challenge now is to discover the physiological role of hormone ouabain. We have recently shown that it modulates cell contacts such as gap junctions, which communicate neighboring cells, as well as tight junctions (TJs), which are one of the two differentiated features of epithelial cells, the other being apical/basolateral polarity. The importance of cell contacts can be hardly overestimated, since the most complex object in the universe, the brain, assembles itself depending on what cells contacts what other(s) how, when, and how is the molecular composition and special arrangement of the contacts involved. In the present chapter, we detail the protocols used to demonstrate the effect of ouabain on the molecular structure and functional properties of one of those cell-cell contacts: the TJ.
哇巴因,一种源自植物的毒素,数世纪以来一直被用于治疗心力衰竭,最近已被证明有内源性对应物,很可能就是哇巴因本身,它具有激素的作用。因此,现在的挑战是发现激素哇巴因的生理作用。我们最近发现它能调节细胞连接,如连接相邻细胞的间隙连接,以及紧密连接(TJ),紧密连接是上皮细胞两个分化特征之一,另一个是顶端/基底外侧极性。细胞连接的重要性再怎么高估都不为过,因为宇宙中最复杂的物体——大脑,是根据细胞如何、何时相互接触以及接触的分子组成和特殊排列方式自我组装的。在本章中,我们详细介绍了用于证明哇巴因对其中一种细胞间连接——紧密连接的分子结构和功能特性影响的实验方案。