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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...
Autores principales: | , , , , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
The Rockefeller University Press
2005
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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. |
format | Text |
id | pubmed-2171740 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2005 |
publisher | The Rockefeller University Press |
record_format | MEDLINE/PubMed |
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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