Open AccessVisualization of Live Primary Cilia Dynamics Using Fluorescence Microscopy
Author(s) -
Ott Carolyn,
LippincottSchwartz Jennifer
Publication year - 2012
Publication title -
current protocols in cell biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.149
H-Index - 38
eISSN - 1934-2616
pISSN - 1934-2500
DOI - 10.1002/0471143030.cb0426s57
Subject(s) - cilium , intraflagellar transport , fluorescence recovery after photobleaching , ciliogenesis , microbiology and biotechnology , microtubule , live cell imaging , biology , dynein , flagellum , cell , biochemistry , membrane , gene
Methods useful for exploring the formation and functions of primary cilia in living cells are described here. First, multiple protocols for visualizing solitary cilia that extend away from the cell body are described. Primary cilia collect, synthesize, and transmit information about the extracellular space into the cell body to promote critical cellular responses. Problems with cilia formation or function can lead to dramatic changes in cell physiology. These methods can be used to assess cilia formation and length, the location of the cilium relative to other cellular structures, and localization of specific proteins to the cilium. The subsequent protocols describe how to quantify movement of fluorescent molecules within the cilium using kymographs, photobleaching, and photoconversion. The microtubules that form the structural scaffold of the cilium are also critical avenues for kinesin and dynein‐mediated movement of proteins within the cilium. Assessing intraflagellar dynamics can provide insight into mechanisms of ciliary‐mediated signal perception and transmission. Curr. Protoc. Cell Biol . 57:4.26.1‐4.26.22. © 2012 by John Wiley & Sons, Inc.