Precursory Research for Embryonic Science and Technology (PRESTO), Japan Science and Technology Agency (JST), Kawaguchi, Saitama, 332-0012, Japan.
Department of Biomolecular Science and Engineering, SANKEN (The Institute of Scientific and Industrial Research), Osaka University, 8-1 Mihogaoka, Ibaraki, 567-0047, Japan.
Commun Biol. 2022 Oct 7;5(1):1065. doi: 10.1038/s42003-022-04023-2.
N-cadherin (NCad) is a classical cadherin that mediates cell-cell interactions in a Ca-dependent manner. NCad participates in various biological processes, from ontogenesis to higher brain functions, though the visualization of NCad interactions in living cells remains limited. Here, we present intensiometric NCad interaction indicators, named INCIDERs, that utilize dimerization-dependent fluorescent proteins. INCIDERs successfully visualize reversible NCad interactions across cells. Compared to FRET-based indicators, INCIDERs have a ~70-fold higher signal contrast, enabling clear identification of NCad interactions. In primary neuronal cells, NCad interactions are visualized between closely apposed processes. Furthermore, visualization of NCad interaction at cell adhesion sites in dense cell populations is achieved by two-photon microscopy. INCIDERs are useful tools in the spatiotemporal investigation of NCad interactions across cells; future research should evaluate the potential of INCIDERs in mapping complex three-dimensional architectures in multi-cellular systems.
N-钙黏蛋白（NCad）是一种经典的钙黏蛋白，以 Ca2+依赖的方式介导细胞-细胞间的相互作用。NCad 参与了从个体发生到高级大脑功能的各种生物学过程，尽管在活细胞中可视化 NCad 相互作用仍然有限。在这里，我们提出了强度计 NCad 相互作用指示剂，命名为 INCIDERs，它利用二聚化依赖性荧光蛋白。INCIDERs 成功地可视化了细胞间可逆的 NCad 相互作用。与基于 FRET 的指示剂相比，INCIDERs 的信号对比度高约 70 倍，能够清晰地识别 NCad 相互作用。在原代神经元细胞中，NCad 相互作用在紧密相邻的突起之间可视化。此外，通过双光子显微镜实现了在密集细胞群体中细胞黏附位点处 NCad 相互作用的可视化。INCIDERs 是在细胞间时空研究 NCad 相互作用的有用工具；未来的研究应该评估 INCIDERs 在绘制多细胞系统中复杂三维结构的潜力。
