Live-Cell Imaging of Dynein-Mediated Cargo Transport in Aspergillus nidulans.
Live-Cell Imaging of Dynein-Mediated Cargo Transport in Aspergillus nidulans.
复制标题
构巢曲霉中动力蛋白介导的货物运输的活细胞成像。
DOI：
10.1007/978-1-0716-2958-1_1
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Xiang,Xin
中科院分区：
文献类型：
--
作者：
Qiu,Rongde;Zhang,Jun;McDaniel,Dennis;Peñalva,MiguelA;Xiang,Xin
丝状真菌已被用于研究细胞质动力蛋白驱动的货物长距离运输，构巢曲霉是研究体内动力蛋白功能和调节的成熟遗传模式生物。在这里，我们描述我们如何成长。nidulansstrains用于活细胞成像，以及我们如何观察早期内体和分泌囊泡的动力蛋白介导的分布。使用倒置显微镜的台上培养箱和培养室，我们可以使用宽视场落射荧光显微镜或新的Zeiss LSM 980（带Airyscan 2）显微镜对自然附着在培养室底部的真菌菌丝进行成像。除了制备用于成像的细胞的方法之外，还提供了用于A.还描述了巢状转变。
Filamentous fungi have been used for studying long-distance transport of cargoes driven by cytoplasmic dynein.Aspergillus nidulansis a well-established genetic model organism used for studying dynein function and regulation in vivo. Here, we describe how we growA. nidulansstrains for live-cell imaging and how we observe the dynein-mediated distribution of early endosomes and secretory vesicles. Using an on-stage incubator and culture chambers for inverted microscopes, we can image fungal hyphae that naturally attach to the bottom of the chambers, using wide-field epifluorescence microscopes or the new Zeiss LSM 980 (with Airyscan 2) microscope. In addition to methods for preparing cells for imaging, a procedure forA. nidulanstransformation is also described.