Department of Molecular Biology, Princeton University, Princeton, NJ 08544USA.
Department of Medicine, Columbia University, New York, NY 10032USA.
Development. 2021 Sep 15;148(18). doi: 10.1242/dev.199695. Epub 2021 Sep 23.
The collective polarization of cellular structures and behaviors across a tissue plane is a near universal feature of epithelia known as planar cell polarity (PCP). This property is controlled by the core PCP pathway, which consists of highly conserved membrane-associated protein complexes that localize asymmetrically at cell junctions. Here, we introduce three new mouse models for investigating the localization and dynamics of transmembrane PCP proteins: Celsr1, Fz6 and Vangl2. Using the skin epidermis as a model, we characterize and verify the expression, localization and function of endogenously tagged Celsr1-3xGFP, Fz6-3xGFP and tdTomato-Vangl2 fusion proteins. Live imaging of Fz6-3xGFP in basal epidermal progenitors reveals that the polarity of the tissue is not fixed through time. Rather, asymmetry dynamically shifts during cell rearrangements and divisions, while global, average polarity of the tissue is preserved. We show using super-resolution STED imaging that Fz6-3xGFP and tdTomato-Vangl2 can be resolved, enabling us to observe their complex localization along junctions. We further explore PCP fusion protein localization in the trachea and neural tube, and discover new patterns of PCP expression and localization throughout the mouse embryo.
细胞结构和行为在组织平面上的集体极化是上皮组织的一个近乎普遍的特征,称为平面细胞极性(PCP)。这种特性由核心 PCP 途径控制,该途径由高度保守的膜相关蛋白复合物组成,这些复合物在细胞连接处不对称定位。在这里,我们引入了三个用于研究跨膜 PCP 蛋白定位和动力学的新的小鼠模型:Celsr1、Fz6 和 Vangl2。我们使用皮肤表皮作为模型,对内源性标记的 Celsr1-3xGFP、Fz6-3xGFP 和 tdTomato-Vangl2 融合蛋白的表达、定位和功能进行了表征和验证。Fz6-3xGFP 在基底表皮祖细胞中的活细胞成像显示,组织的极性不是随着时间固定的。相反,不对称性在细胞重排和分裂过程中动态移动,而组织的整体平均极性得以保持。我们使用超分辨率 STED 成像表明,Fz6-3xGFP 和 tdTomato-Vangl2 可以被分辨,使我们能够观察它们在连接处的复杂定位。我们进一步探索了 PCP 融合蛋白在气管和神经管中的定位,并在整个小鼠胚胎中发现了新的 PCP 表达和定位模式。