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Multi-color live-cell fluorescence imaging of primary ciliary membrane assembly and dynamics

  1. Author:
    Lu,Quanlong
    Westlake,Christopher

  2. Author Address

    Laboratory of Cell and Developmental Signaling, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Frederick, MD, United States. Electronic address: quanlong.lu@nih.gov., Laboratory of Cell and Developmental Signaling, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Frederick, MD, United States. Electronic address: chris.westlake@nih.gov.,
    1. Year: 2023
    2. Epub Date: 2023 03 27
  2. Journal: Methods in Cell Biology
    1. 176
    2. Pages: 235-250
  4. Type of Article: Article
  1. Abstract:

    The ciliary membrane is continuous with the plasma membrane but has distinct lipid and protein composition, which is key to defining the function of the primary cilium. Ciliary membranes dynamically assemble and disassemble in association with the cell cycle and directly transmit signals and molecules through budding membranes. Various imaging approaches have greatly advanced the understanding of the ciliary membrane function. In particular, fluorescence live-cell imaging has revealed important insights into the dynamics of ciliary membrane assembly by monitoring the changes of fluorescent-tagged ciliary proteins. Protein dynamics can be tracked simultaneously using multi-color live cell imaging by coupling ciliary-associated factors with different colored fluorescent tags. Ciliary membrane and membrane associated-proteins such as Smoothened, 5-HTr6, SSTR3, Rab8a, and Arl13b have been used to track ciliary membranes and centriole proteins like Centrin1/2, CEP164, and CEP83 are often used to mark the ciliary basal body. Here, we describe a method for studying ciliogenesis membrane dynamics using spinning disk confocal live-cell imaging. Copyright © 2023 Elsevier Inc. All rights reserved.

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External Sources

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  2. DOI: 10.1016/bs.mcb.2023.02.001
  3. PMID: 37164540
  4. PII : S0091-679X(23)00045-6

Library Notes

  1. Fiscal Year: FY2022-2023
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