Immunoelectron Microscopy of Vesicle Transport to the Primary Cilium of Photoreceptor Cells

Publication type:
Chapter
Metadata:
Autoren
  • Tina Sedmak
  • Elisabeth Sehn
  • Uwe Wolfrum
Autoren-URL
https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=fis-test-1&SrcAuth=WosAPI&KeyUT=WOS:000272975500014&DestLinkType=FullRecord&DestApp=WOS_CPL
DOI
10.1016/S0091-679X(08)94013-9
ISBN-13
978-0-12-375024-2
Paginierung
259 - +
Buchtitel
PRIMARY CILIA
Datum der Veröffentlichung
2009
Status
Published
Titel
Immunoelectron Microscopy of Vesicle Transport to the Primary Cilium of Photoreceptor Cells
Ausgabe der Zeitschrift
94

Data source: Web of Science (Lite)

Other metadata sources:
Autoren
  • Tina Sedmak
  • Elisabeth Sehn
  • Uwe Wolfrum
DOI
10.1016/s0091-679x(08)94013-9
Paginierung
259 - 272
Buchtitel
Methods in Cell Biology
Datum der Veröffentlichung
2009
Status
Published
Herausgeber
Elsevier
Herausgeber URL
http://dx.doi.org/10.1016/s0091-679x(08)94013-9
Datum der Datenerfassung
2019
Titel
Immunoelectron Microscopy of Vesicle Transport to the Primary Cilium of Photoreceptor Cells

Data source: Crossref

Abstract
Cilia are organelles of high structural complexity. Since the biosynthetic machinery is absent from cilia all their molecular components must be synthesized in organelles of the cytoplasm and subsequently transported to the cilium. Ciliary cargos are thought to be translocated in the membrane of transport vesicles or association with these vesicles to the base of the cilium where the vesicles fuse with the periciliary target membrane for further delivery of their cargo into the ciliary compartment by the intraflagellar transport (IFT). Here we describe a modified preembedding labeling method as an alternative technique to conventional postembedding methods eligible for analyses of ciliary cargo vesicles and the distribution of ciliary molecules in subciliary compartments for immunoelectron microscopy. The preembedding labeling method preserves the antigenicity of ciliary antigens and its application reveals differential localization of individual IFT proteins in vertebrate photoreceptor cilia. Since membrane vesicles are conserved, the preembedding protocol additionally allows the identification of ciliary cargo vesicles by immunolabeling of individual IFT proteins and ciliary targeting molecules in ciliary photoreceptor cells. These results do not only confirm the central function of IFT molecules in ciliary transport, but further strengthen their role in transport processes in the cytoplasm. Furthermore, evidence for different alternative transport routes of cargo vesicles directed to different target membranes is gathered.
Addresses
  • Department of Cell and Matrix Biology, Institute of Zoology, Johannes Gutenberg University, Mainz D-55099, Germany.
Autoren
  • Tina Sedmak
  • Elisabeth Sehn
  • Uwe Wolfrum
DOI
10.1016/s0091-679x(08)94013-9
Open access
false
Schlüsselwörter
  • Cilia
  • Transport Vesicles
  • Animals
  • Microscopy, Immunoelectron
  • Tissue Fixation
  • Photoreceptor Cells
Medium
Print-Electronic
Online publication date
2009
Paginierung
259 - 272
Datum der Veröffentlichung
2009
Status
Published
Datum der Datenerfassung
2010
Titel
Immunoelectron microscopy of vesicle transport to the primary cilium of photoreceptor cells.
Ausgabe der Zeitschrift
94

Data source: Europe PubMed Central

Abstract
Cilia are organelles of high structural complexity. Since the biosynthetic machinery is absent from cilia all their molecular components must be synthesized in organelles of the cytoplasm and subsequently transported to the cilium. Ciliary cargos are thought to be translocated in the membrane of transport vesicles or association with these vesicles to the base of the cilium where the vesicles fuse with the periciliary target membrane for further delivery of their cargo into the ciliary compartment by the intraflagellar transport (IFT). Here we describe a modified preembedding labeling method as an alternative technique to conventional postembedding methods eligible for analyses of ciliary cargo vesicles and the distribution of ciliary molecules in subciliary compartments for immunoelectron microscopy. The preembedding labeling method preserves the antigenicity of ciliary antigens and its application reveals differential localization of individual IFT proteins in vertebrate photoreceptor cilia. Since membrane vesicles are conserved, the preembedding protocol additionally allows the identification of ciliary cargo vesicles by immunolabeling of individual IFT proteins and ciliary targeting molecules in ciliary photoreceptor cells. These results do not only confirm the central function of IFT molecules in ciliary transport, but further strengthen their role in transport processes in the cytoplasm. Furthermore, evidence for different alternative transport routes of cargo vesicles directed to different target membranes is gathered.
Autoren
  • Tina Sedmak
  • Elisabeth Sehn
  • Uwe Wolfrum
Autoren-URL
https://www.ncbi.nlm.nih.gov/pubmed/20362095
DOI
10.1016/S0091-679X(08)94013-9
Schlüsselwörter
  • Animals
  • Cilia
  • Microscopy, Immunoelectron
  • Photoreceptor Cells
  • Tissue Fixation
  • Transport Vesicles
Paginierung
259 - 272
Datum der Veröffentlichung
2009
Status
Published
Datum, an dem der Datensatz öffentlich gemacht wurde
2010
Titel
Immunoelectron microscopy of vesicle transport to the primary cilium of photoreceptor cells.
Ausgabe der Zeitschrift
94

Data source: PubMed

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