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Cilia pp 35-51 | Cite as

Morphological and Functional Characterization of the Ciliary Pocket by Electron and Fluorescence Microscopy

  • Rania Ghossoub
  • Louise Lindbæk
  • Anahi Molla-Herman
  • Alain Schmitt
  • Søren Tvorup ChristensenEmail author
  • Alexandre BenmerahEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1454)

Abstract

In many vertebrate cell types, the proximal part of the primary cilium is positioned within an invagination of the plasma membrane known as the ciliary pocket. Recent evidence points to the conclusion that the ciliary pocket comprises a unique site for exocytosis and endocytosis of ciliary proteins, which regulates the spatiotemporal trafficking of receptors into and out of the cilium to control its sensory function. In this chapter, we provide methods based on electron microscopy, 3D reconstruction of fluorescence images as well as live cell imaging suitable for investigating processes associated with endocytosis at the ciliary pocket.

Key words

Primary cilia Ciliary pocket Clathrin-dependent endocytosis Early endosomes Cellular signaling 

Notes

Acknowledgments

We would like to thank Pierre Bourdoncle (Cellular imaging Facility, Cochin Institute) for his precious help on deconvolution and 3D reconstruction of fluorescence images on RPE1 cells. This work was supported by the University of Copenhagen Excellence Programme for Interdisciplinary Research and the Danish Council for Independent Research (1331-00254).

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Rania Ghossoub
    • 1
  • Louise Lindbæk
    • 2
  • Anahi Molla-Herman
    • 3
    • 9
  • Alain Schmitt
    • 4
    • 5
    • 6
  • Søren Tvorup Christensen
    • 2
    Email author
  • Alexandre Benmerah
    • 7
    • 8
    Email author
  1. 1.Centre de Recherche en Cancérologie de Marseille (CRCM), Inserm, U1068-CNRS UMR7258, Institut Paoli-CalmettesAix-Marseille UniversitéMarseilleFrance
  2. 2.Department of BiologyUniversity of CopenhagenCopenhagenDenmark
  3. 3.Department of Genetics and Developmental BiologyInstitut CurieParisFrance
  4. 4.INSERM, U1016Institut CochinParisFrance
  5. 5.CNRS, UMR8104ParisFrance
  6. 6.Université Paris DescartesParisFrance
  7. 7.INSERM, UMR-1163Laboratory of Inherited Kidney DiseasesParisFrance
  8. 8.Université Paris Descartes-Sorbonne Paris CitéImagine InstitutParisFrance
  9. 9.CNRS, UMRParisFrance

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