Document Detail


Estimation of PC12 cell numbers with acid phosphatase assay and mitochondrial dehydrogenase assay: dopamine interferes with assay based on tetrazolium.
MedLine Citation:
PMID:  9928836     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Both the acid-phosphatase and mitochondrial dehydrogenase assay have been used to quantify cell numbers. The commonly used acid-phosphatase assay uses p-nitrophenyl phosphate as a substrate, while the mitochondrial dehydrogenase assay is based on the conversion of tetrazolium to formazan. Our experimental results showed that the former assay was more sensitive in detecting small numbers of PC12 cells (200-10,000 cells/well), whereas the latter was useful for larger numbers of cells (2000-40,000 cells/well). The number of PC12 cells decreased after dopamine treatment, according to the acid-phosphatase assay and by direct cell counts under a light microscope. However, the optical densities measured by the mitochondrial dehydrogenase assay increased after dopamine treatment. We tried to clarify discrepancies between the two assays, since dopamine is an important neurotransmitter and both assays are commonly used to estimate cell numbers. To elucidate the interference between dopamine and tetrazolium salt, cell-free control experiments were performed. Dopamine and other catecholamines (adrenaline and noradrenaline) reacted with tetrazolium and, thus, produced a false positive reaction in the assay. We therefore conclude that the tetrazolium assay is not a suitable method for evaluating the number of catecholamine-treated cells, while the acid-phosphatase assay is reliable and sensitive.
Authors:
F Si; S H Shin; A Biedermann; G M Ross
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Experimental brain research     Volume:  124     ISSN:  0014-4819     ISO Abbreviation:  Exp Brain Res     Publication Date:  1999 Jan 
Date Detail:
Created Date:  1999-03-29     Completed Date:  1999-03-29     Revised Date:  2013-12-13    
Medline Journal Info:
Nlm Unique ID:  0043312     Medline TA:  Exp Brain Res     Country:  GERMANY    
Other Details:
Languages:  eng     Pagination:  145-50     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Acid Phosphatase / analysis*
Animals
Antioxidants / pharmacology
Ascorbic Acid / pharmacology
Cell Count / methods
Clinical Enzyme Tests / standards
Dopamine / analysis*,  metabolism
Epinephrine / analysis
False Positive Reactions
Mitochondria / drug effects,  enzymology
Neurons / chemistry,  cytology,  enzymology*
Norepinephrine / analysis
Oxidoreductases / analysis*
PC12 Cells
Rats
Sensitivity and Specificity
Tetrazolium Salts
Chemical
Reg. No./Substance:
0/Antioxidants; 0/Tetrazolium Salts; EC 1.-/Oxidoreductases; EC 3.1.3.2/Acid Phosphatase; PQ6CK8PD0R/Ascorbic Acid; VTD58H1Z2X/Dopamine; X4W3ENH1CV/Norepinephrine; YKH834O4BH/Epinephrine

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


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