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对斑马鱼中的纤毛进行成像。

Imaging cilia in zebrafish.

作者信息

Jaffe Kimberly M, Thiberge Stephan Y, Bisher Margaret E, Burdine Rebecca D

机构信息

Department of Molecular Biology, Princeton University, Princeton, New Jersey 08544, USA.

出版信息

Methods Cell Biol. 2010;97:415-35. doi: 10.1016/S0091-679X(10)97022-2.

Abstract

Research focused on cilia as extremely important cellular organelles has flourished in recent years. A thorough understanding of cilia regulation and function is critical, as disruptions of cilia structure and/or function have been linked to numerous human diseases and disorders. The tropical freshwater zebrafish is an excellent model organism in which to study cilia structure and function. We can readily image cilia and their motility in embryonic structures including Kupffer's vesicle during somite stages and the pronephros from 1 day postfertilization onward. Here, we describe how to image cilia by whole-mount immunofluorescence, transverse cryosection/immunohistochemistry, and transmission electron microscopy. We also describe how to obtain videos of cilia motility in living embryos.

摘要

近年来,针对作为极其重要的细胞器的纤毛的研究蓬勃发展。深入了解纤毛的调节和功能至关重要,因为纤毛结构和/或功能的破坏与众多人类疾病和病症有关。热带淡水斑马鱼是研究纤毛结构和功能的优秀模式生物。我们可以很容易地在胚胎结构中观察纤毛及其运动,包括体节期的库普弗囊泡以及受精后1天起的前肾。在这里,我们描述了如何通过整体免疫荧光、横向冷冻切片/免疫组织化学和透射电子显微镜对纤毛进行成像。我们还描述了如何获取活胚胎中纤毛运动的视频。

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