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

Methods Mol Biol. 2016:1454:35-51. doi: 10.1007/978-1-4939-3789-9_3.

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.

Keywords: Cellular signaling; Ciliary pocket; Clathrin-dependent endocytosis; Early endosomes; Primary cilia.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Cilia / metabolism*
  • Cilia / ultrastructure*
  • Clathrin / metabolism
  • Clathrin-Coated Vesicles / metabolism
  • Endocytosis
  • Endosomes / metabolism
  • Gene Expression
  • Genes, Reporter
  • Humans
  • Imaging, Three-Dimensional
  • Microscopy, Electron*
  • Microscopy, Fluorescence*
  • Organogenesis / genetics
  • Protein Transport
  • Retinal Pigment Epithelium
  • Signal Transduction
  • Transferrin / metabolism
  • Transforming Growth Factor beta1 / metabolism

Substances

  • Clathrin
  • Transferrin
  • Transforming Growth Factor beta1