Document Detail


Assay of mitochondrial functions by resazurin in vitro.
MedLine Citation:
PMID:  15000895     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
AIM: To study the mechanism of resazurin as indicator of mitochondrial function and to develop a rapid and sensitive assay for measuring metabolic activity of isolated mitochondria from rat liver in vitro. METHODS: The screening was carried out on 96-well microtitre plates by monitoring fluorescence intensity of resazurin reduced by mitochondria. Experimental conditions were optimized and influences of several inhibitors on mitochondrial function were observed. RESULTS: Fluorescence intensity increased in a linear manner when the mitochondrial protein concentration from 5 to 50 microg protein per well was incubated with resazurin (5 micromol/L) during 230 min period at 37 degrees C. Edetic acid could promote the reduction of resazurin in mitochondria. The fluorescence intensity decreased greatly after pretreatment with NaN3, antimycin A, carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP), and oligomycin compared with the control. However, the typical complex I inhibitor, rotenone enhanced the fluorescence intensity without mitochondria. CONCLUSION: Using resazurin to determine mitochondrial function is sensitive, inexpensive and could be easily automated for high throughput screening.
Authors:
Hai-xia Zhang; Guan-hua Du; Jun-tian Zhang
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Publication Detail:
Type:  In Vitro; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Acta pharmacologica Sinica     Volume:  25     ISSN:  1671-4083     ISO Abbreviation:  Acta Pharmacol. Sin.     Publication Date:  2004 Mar 
Date Detail:
Created Date:  2004-03-05     Completed Date:  2004-08-06     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  100956087     Medline TA:  Acta Pharmacol Sin     Country:  China    
Other Details:
Languages:  eng     Pagination:  385-9     Citation Subset:  IM    
Affiliation:
Institute of Materia Medica, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing 100050, China.
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MeSH Terms
Descriptor/Qualifier:
Animals
Antimycin A / pharmacology
Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone / pharmacology
Edetic Acid / pharmacology
Fluorescence
Male
Mitochondria, Liver / drug effects,  metabolism*
Mitochondrial Proteins / metabolism
Nitrates / pharmacology
Oligomycins / pharmacology
Oxazines / metabolism*
Rats
Rats, Sprague-Dawley
Rotenone / pharmacology
Xanthenes*
Chemical
Reg. No./Substance:
0/Mitochondrial Proteins; 0/Nitrates; 0/Oligomycins; 0/Oxazines; 0/Xanthenes; 370-86-5/Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone; 550-82-3/resazurin; 60-00-4/Edetic Acid; 642-15-9/Antimycin A; 7631-99-4/sodium nitrate; 83-79-4/Rotenone

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


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