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Ultrastructural identification of uncoated caveolin-independent early endocytic vehicles

Using quantitative light microscopy and a modified immunoelectron microscopic technique, we have characterized the entry pathway of the cholera toxin binding subunit (CTB) in primary embryonic fibroblasts. CTB trafficking to the Golgi complex was identical in caveolin-1null (Cav1−/−) mouse embryonic...

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Autores principales: Kirkham, Matthew, Fujita, Akikazu, Chadda, Rahul, Nixon, Susan J., Kurzchalia, Teymuras V., Sharma, Deepak K., Pagano, Richard E., Hancock, John F., Mayor, Satyajit, Parton, Robert G.
Formato: Texto
Lenguaje:English
Publicado: The Rockefeller University Press 2005
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2171740/
https://www.ncbi.nlm.nih.gov/pubmed/15668297
http://dx.doi.org/10.1083/jcb.200407078
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author Kirkham, Matthew
Fujita, Akikazu
Chadda, Rahul
Nixon, Susan J.
Kurzchalia, Teymuras V.
Sharma, Deepak K.
Pagano, Richard E.
Hancock, John F.
Mayor, Satyajit
Parton, Robert G.
author_facet Kirkham, Matthew
Fujita, Akikazu
Chadda, Rahul
Nixon, Susan J.
Kurzchalia, Teymuras V.
Sharma, Deepak K.
Pagano, Richard E.
Hancock, John F.
Mayor, Satyajit
Parton, Robert G.
author_sort Kirkham, Matthew
collection PubMed
description Using quantitative light microscopy and a modified immunoelectron microscopic technique, we have characterized the entry pathway of the cholera toxin binding subunit (CTB) in primary embryonic fibroblasts. CTB trafficking to the Golgi complex was identical in caveolin-1null (Cav1−/−) mouse embryonic fibroblasts (MEFs) and wild-type (WT) MEFs. CTB entry in the Cav1−/− MEFs was predominantly clathrin and dynamin independent but relatively cholesterol dependent. Immunoelectron microscopy was used to quantify budded and surface-connected caveolae and to identify noncaveolar endocytic vehicles. In WT MEFs, a small fraction of the total Cav1-positive structures were shown to bud from the plasma membrane (2% per minute), and budding increased upon okadaic acid or lactosyl ceramide treatment. However, the major carriers involved in initial entry of CTB were identified as uncoated tubular or ring-shaped structures. These carriers contained GPI-anchored proteins and fluid phase markers and represented the major vehicles mediating CTB uptake in both WT and caveolae-null cells.
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spelling pubmed-21717402008-03-05 Ultrastructural identification of uncoated caveolin-independent early endocytic vehicles Kirkham, Matthew Fujita, Akikazu Chadda, Rahul Nixon, Susan J. Kurzchalia, Teymuras V. Sharma, Deepak K. Pagano, Richard E. Hancock, John F. Mayor, Satyajit Parton, Robert G. J Cell Biol Research Articles Using quantitative light microscopy and a modified immunoelectron microscopic technique, we have characterized the entry pathway of the cholera toxin binding subunit (CTB) in primary embryonic fibroblasts. CTB trafficking to the Golgi complex was identical in caveolin-1null (Cav1−/−) mouse embryonic fibroblasts (MEFs) and wild-type (WT) MEFs. CTB entry in the Cav1−/− MEFs was predominantly clathrin and dynamin independent but relatively cholesterol dependent. Immunoelectron microscopy was used to quantify budded and surface-connected caveolae and to identify noncaveolar endocytic vehicles. In WT MEFs, a small fraction of the total Cav1-positive structures were shown to bud from the plasma membrane (2% per minute), and budding increased upon okadaic acid or lactosyl ceramide treatment. However, the major carriers involved in initial entry of CTB were identified as uncoated tubular or ring-shaped structures. These carriers contained GPI-anchored proteins and fluid phase markers and represented the major vehicles mediating CTB uptake in both WT and caveolae-null cells. The Rockefeller University Press 2005-01-31 /pmc/articles/PMC2171740/ /pubmed/15668297 http://dx.doi.org/10.1083/jcb.200407078 Text en Copyright © 2005, The Rockefeller University Press This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).
spellingShingle Research Articles
Kirkham, Matthew
Fujita, Akikazu
Chadda, Rahul
Nixon, Susan J.
Kurzchalia, Teymuras V.
Sharma, Deepak K.
Pagano, Richard E.
Hancock, John F.
Mayor, Satyajit
Parton, Robert G.
Ultrastructural identification of uncoated caveolin-independent early endocytic vehicles
title Ultrastructural identification of uncoated caveolin-independent early endocytic vehicles
title_full Ultrastructural identification of uncoated caveolin-independent early endocytic vehicles
title_fullStr Ultrastructural identification of uncoated caveolin-independent early endocytic vehicles
title_full_unstemmed Ultrastructural identification of uncoated caveolin-independent early endocytic vehicles
title_short Ultrastructural identification of uncoated caveolin-independent early endocytic vehicles
title_sort ultrastructural identification of uncoated caveolin-independent early endocytic vehicles
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2171740/
https://www.ncbi.nlm.nih.gov/pubmed/15668297
http://dx.doi.org/10.1083/jcb.200407078
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