Paniagua Antonio E, Segurado Alicia, Dolón Jorge F, Esteve-Rudd Julián, Velasco Almudena, Williams David S, Lillo Concepción
Institute of Neurosciences of Castilla y León, IBSAL, Cell Biology and Pathology, University of Salamanca, Salamanca, Spain.
Stein Eye Institute and Department of Ophthalmology, David Geffen School of Medicine at UCLA, Los Angeles, CA, United States.
Front Cell Dev Biol. 2021 Jun 28;9:701853. doi: 10.3389/fcell.2021.701853. eCollection 2021.
Apicobasal polarity is essential for epithelial cell function, yet the roles of different proteins in its completion is not fully understood. Here, we have studied the role of the polarity protein, CRB2, in human retinal pigment epithelial (RPE) cells during polarization , and in mature murine RPE cells . After establishing a simplified protocol for the culture of human fetal RPE cells, we studied the temporal sequence of the expression and localization of polarity and cell junction proteins during polarization in these epithelial cells. We found that CRB2 plays a key role in tight junction maintenance as well as in cell cycle arrest. In addition, our studies show that the knockdown of CRB2 in the RPE affects to the distribution of different apical polarity proteins and results in perturbed retinal homeostasis, manifested by the invasion of activated microglial cells into the subretinal space. Together our results demonstrate that CRB2 is a key protein for the development and maintenance of a polarized epithelium.
顶-基极性对于上皮细胞功能至关重要,然而不同蛋白质在其形成过程中的作用尚未完全明确。在此,我们研究了极性蛋白CRB2在人视网膜色素上皮(RPE)细胞极化过程中以及在成熟小鼠RPE细胞中的作用。在建立了一种简化的人胎儿RPE细胞培养方案后,我们研究了这些上皮细胞在极化过程中极性和细胞连接蛋白表达及定位的时间顺序。我们发现CRB2在紧密连接维持以及细胞周期停滞中起关键作用。此外,我们的研究表明,RPE细胞中CRB2的敲低会影响不同顶端极性蛋白的分布,并导致视网膜内环境稳态受到干扰,表现为活化的小胶质细胞侵入视网膜下间隙。我们的研究结果共同表明,CRB2是极化上皮发育和维持的关键蛋白。