Zhang Yong, Huang Guangming, Shornick Laurie P, Roswit William T, Shipley J Michael, Brody Steven L, Holtzman Michael J
Department of Medicine, Washington University School of Medicine, St. Louis, MO 63110-1093, USA.
Am J Respir Cell Mol Biol. 2007 May;36(5):515-9. doi: 10.1165/rcmb.2006-0475RC. Epub 2007 Jan 25.
Ciliated airway epithelial cells are critical for mucosal barrier function, including host defense against pathogens. This cell population is often the primary target and thereby the first line of defense against many common respiratory viruses. It is also the precursor for mucous cells and thereby promotes mucociliary clearance of infectious and other noxious agents. Cells with motile cilia in other organs (e.g., brain and reproductive organs) may also have roles in development and reproduction. However, definitive proof of ciliated cell function is hampered by the lack of strategies to specifically target this cell population for loss of function in vivo. To this end, cell type-specific gene promoters have been combined with the Cre/LoxP system to disrupt genes in airway and alveolar epithelial cell populations expressing surfactant protein C (SP-C) or Clara cell secretory protein (CCSP). By contrast, an analogous system to disrupt gene function in ciliated airway epithelial cells was still needed. Here we report the generation and analysis of mouse lines with a FOXJ1 promoter driving the Cre recombinase and show that this system mediates genomic recombination specifically in ciliated cells. The pattern of recombination recapitulates endogenous FOXJ1 promoter function, being restricted to ciliated cells present in pulmonary airways as well as choroid plexus, ependyma, oviduct, and testis. This transgenic mouse system thereby offers a new strategy for specific knockouts of genes in ciliated cells. It should prove extremely useful for defining ciliated cell function in airway mucosal immunity as well as development and reproduction.
纤毛气道上皮细胞对于黏膜屏障功能至关重要,包括宿主抵御病原体。这一细胞群体常常是主要靶点,因此是抵御许多常见呼吸道病毒的第一道防线。它也是黏液细胞的前体,从而促进对传染性及其他有害物质的黏液纤毛清除。其他器官(如脑和生殖器官)中具有活动纤毛的细胞在发育和生殖中可能也发挥作用。然而,由于缺乏在体内特异性靶向这一细胞群体使其功能丧失的策略,纤毛细胞功能的确切证据受到了阻碍。为此,细胞类型特异性基因启动子已与Cre/LoxP系统相结合,以破坏表达表面活性蛋白C(SP-C)或克拉拉细胞分泌蛋白(CCSP)的气道和肺泡上皮细胞群体中的基因。相比之下,仍需要一种类似的系统来破坏纤毛气道上皮细胞中的基因功能。在此,我们报告了由FOXJ1启动子驱动Cre重组酶的小鼠品系的产生和分析,并表明该系统在纤毛细胞中特异性介导基因组重组。重组模式重现了内源性FOXJ1启动子的功能,仅限于肺气道以及脉络丛、室管膜、输卵管和睾丸中的纤毛细胞。这种转基因小鼠系统因此为在纤毛细胞中特异性敲除基因提供了一种新策略。它对于确定纤毛细胞在气道黏膜免疫以及发育和生殖中的功能应该极其有用。