关键词: Cilia length Gene knock-down Immunofluorescence Primary cilia Renal epithelial cells Scanning electron microscopy

Mesh : Animals Cells, Cultured Cilia / metabolism ultrastructure Epithelial Cells / cytology metabolism ultrastructure Fluorescent Antibody Technique / instrumentation methods Gene Knockdown Techniques / instrumentation methods HEK293 Cells Histocytological Preparation Techniques / instrumentation methods Humans Intravital Microscopy / instrumentation methods Kidney / cytology metabolism ultrastructure Mice Microscopy, Electron, Scanning / instrumentation methods Microscopy, Fluorescence / instrumentation methods RNA, Small Interfering Signal Transduction

来  源:   DOI:10.1016/bs.mcb.2019.04.008   PDF(Sci-hub)   PDF(Pubmed)

Abstract:
Primary cilia are singular, sensory organelles that extend from the plasma membrane of most quiescent mammalian cells. These slender, microtubule-based organelles receive and transduce extracellular cues and regulate signaling pathways. Primary cilia are critical to the development and function of many tissue types, and mutation of ciliary genes causes multi-system disorders, termed ciliopathies. Notably, renal cystic disease is one of the most common clinical features of ciliopathies, highlighting a central role for primary cilia in the kidney. Additionally, acute kidney injury and chronic kidney disease are associated with altered primary cilia lengths on renal epithelial cells, suggesting ciliary dynamics and renal physiology are linked. Here we describe methods to examine primary cilia in kidney tissue and in cultured renal cells. We include immunofluorescence and scanning electron microscopy to determine ciliary localization of proteins and cilia structure. Further, we detail cellular assays to measure cilia assembly and disassembly, which regulate cilia length.
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