Document Detail


Sphingolipid trafficking and purification in Chlamydia trachomatis-infected cells.
MedLine Citation:
PMID:  23184593     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Chlamydia trachomatis is an obligate intracellular human pathogen, which lacks a system that allows genetic manipulation. Therefore, chlamydial researchers must manipulate the host cell to better understand chlamydial biology. Host-derived lipid acquisition is critical for chlamydial survival within the host. Hence, the ability to track and purify sphingolipids in/from chlamydial infected cells has become an integral part of pivotal studies in chlamydial biology. This unit outlines protocols that provide details about labeling eukaryotic cells with exogenous lipids to examine Golgi-derived lipid trafficking to the chlamydial inclusion and then performing imaging studies or lipid extractions for quantification. Details are provided to allow these protocols to be applied to subconfluent, polarized, or siRNA knockdown cells. In addition, one will find important experimental design considerations and techniques. These methods are powerful tools to aid in the understanding of mechanisms, which allow C. trachomatis to manipulate and usurp host cell trafficking pathways.
Authors:
Elizabeth R Moore
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Current protocols in microbiology     Volume:  Chapter 11     ISSN:  1934-8533     ISO Abbreviation:  Curr Protoc Microbiol     Publication Date:  2012 Nov 
Date Detail:
Created Date:  2012-11-27     Completed Date:  2013-04-26     Revised Date:  2013-11-06    
Medline Journal Info:
Nlm Unique ID:  101257113     Medline TA:  Curr Protoc Microbiol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  Unit 11A.2.     Citation Subset:  IM    
Copyright Information:
© 2012 by John Wiley & Sons, Inc.
Affiliation:
Division of Basic Biomedical Sciences, Sanford School of Medicine, University of South Dakota, Vermillion, South Dakota, USA.
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MeSH Terms
Descriptor/Qualifier:
Biological Transport
Cell Culture Techniques / methods*
Cell Tracking / methods*
Chlamydia Infections / metabolism,  microbiology*
Chlamydia trachomatis / chemistry,  isolation & purification,  metabolism*
Humans
Sphingolipids / chemistry*,  isolation & purification*,  metabolism
Staining and Labeling / methods*
Grant Support
ID/Acronym/Agency:
K22 AI089856/AI/NIAID NIH HHS; K22AI089856/AI/NIAID NIH HHS
Chemical
Reg. No./Substance:
0/Sphingolipids
Comments/Corrections

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