Document Detail

A resealed-cell system for analyzing pathogenic intracellular events: perturbation of endocytic pathways under diabetic conditions.
MedLine Citation:
PMID:  22952896     Owner:  NLM     Status:  MEDLINE    
Cell-based assay systems that can serve as cellular models of aberrant function in pathogenic organs would be novel and useful tools for screening drugs and clarifying the molecular mechanisms of various diseases. We constructed model cells that replicated the conditions in diabetic hepatocytes by using the cell resealing technique, which enables the exchange of cytosol. The plasma membrane of HeLa cells was permeabilized with the streptococcal toxin streptolysin O, and cytosol that had been prepared from wild-type or db/db diabetic mice was introduced into the resulting semi-intact cells. By resealing the plasma membrane by exposure to Ca(2+), we created WT or Db model cells, in which the cytosolic conditions replicated those of healthy or diabetic liver. Interestingly, phosphorylation of p38 MAPK was promoted, whereas the level of endosomal phosphatidylinositol-3-phosphate was decreased, in Db cells. We investigated several endocytic pathways in WT and Db cells, and found that retrograde endosome-to-Golgi transport was delayed in a p38 MAPK-dependent manner in Db cells. Furthermore, the degradation pathway of the EGF receptor from endosomes to lysosomes was enhanced in Db cells, and this did not depend on the activation of p38 MAPK. The disease model cell system should become a powerful tool for the detection of aberrant processes in cells under pathogenic conditions and for therapeutic applications.
Fumi Kano; Daiki Nakatsu; Yoshiyuki Noguchi; Akitsugu Yamamoto; Masayuki Murata
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2012-08-29
Journal Detail:
Title:  PloS one     Volume:  7     ISSN:  1932-6203     ISO Abbreviation:  PLoS ONE     Publication Date:  2012  
Date Detail:
Created Date:  2012-09-06     Completed Date:  2013-02-07     Revised Date:  2013-07-12    
Medline Journal Info:
Nlm Unique ID:  101285081     Medline TA:  PLoS One     Country:  United States    
Other Details:
Languages:  eng     Pagination:  e44127     Citation Subset:  IM    
Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Meguro-ku, Tokyo, Japan.
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MeSH Terms
Bacterial Proteins / pharmacology
Calcium Chloride / pharmacology
Cell Membrane Permeability / drug effects
Cholera Toxin / metabolism
Dextrans / metabolism
Diabetes Mellitus / metabolism,  pathology*
Endocytosis / drug effects*
Endosomes / drug effects,  metabolism,  ultrastructure
Fluoresceins / metabolism
Golgi Apparatus / drug effects,  metabolism,  ultrastructure
HeLa Cells
Intracellular Space / drug effects,  metabolism*
Liver / drug effects,  metabolism,  pathology
Models, Biological
Molecular Weight
Phosphatidylinositol Phosphates / metabolism
Protein Transport / drug effects
Proteolysis / drug effects
Receptor, Epidermal Growth Factor / metabolism
Streptolysins / pharmacology
Transferrin / metabolism
p38 Mitogen-Activated Protein Kinases / metabolism
Reg. No./Substance:
0/Bacterial Proteins; 0/Fluoresceins; 0/Phosphatidylinositol Phosphates; 0/Streptolysins; 0/Transferrin; 0/fluorescein-dextran; 0/phosphatidylinositol 3-phosphate; 0/streptolysin O; 10043-52-4/Calcium Chloride; 9004-54-0/Dextrans; 9012-63-9/Cholera Toxin; EC, Epidermal Growth Factor; EC Mitogen-Activated Protein Kinases

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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