Document Detail


Free cytosolic Ca2+ measurements with fluorine labelled indicators using 19FNMR.
MedLine Citation:
PMID:  3874697     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Characterisation by 19F NMR of fluorine-labelled indicators of cytosolic free Ca2+ concentration (by 5FBAPTA) and pH (by Fquene) is described, together with the techniques used to load the cell suspensions with the indicators for NMR spectroscopy. Useful features of the 19F NMR indicators include direct identification of the intracellular cation bound to the indicators, internal calibration of [Ca]i and pHi from the spectra, and simultaneous measurements of two or more indicators in the same cell suspension. Perturbations of cellular functions by 5FBAPTA and quin 2 are very similar, but vary widely in different cell systems. The [Ca]i and pHi responses of normal and transformed cells to mitogens and growth factors in serum are compared with data from similar experiments using fluorescence indicators. The only major discrepancy in [Ca]i measurements using the two independent assays was observed in Ehrlich ascites tumour cells. These cells have a high intracellular Zn2+ content which substantially quenches the quin 2 fluorescence, but does not affect [Ca]i measurements by 5FBAPTA. The Zn2+ present in the cells is detected as a separate response in the 5FBAPTA spectrum. The time course of the Ca signal in 2H3 cells stimulated by antigen to release histamine by exocytosis has been defined using 5FBAPTA and quin 2. Extension of the 19F NMR technique to [Ca] i and pHi measurements in perfused organs is illustrated in rat heart and responses to pharmacological agents are demonstrated. Developments in prospect to improve sensitivity and to measure [Na]i with a new family of indicators are outlined.
Authors:
J C Metcalfe; T R Hesketh; G A Smith
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Cell calcium     Volume:  6     ISSN:  0143-4160     ISO Abbreviation:  Cell Calcium     Publication Date:  1985 Apr 
Date Detail:
Created Date:  1985-08-23     Completed Date:  1985-08-23     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8006226     Medline TA:  Cell Calcium     Country:  SCOTLAND    
Other Details:
Languages:  eng     Pagination:  183-95     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Aminoquinolines / diagnostic use
Aniline Compounds / diagnostic use*
Animals
Calcium / analysis*
Carcinoma, Ehrlich Tumor / analysis
Cytosol / analysis*
Exocytosis
Fluorescent Dyes / diagnostic use*
Fluorine
Hydrogen-Ion Concentration
Leukemia / metabolism
Magnetic Resonance Spectroscopy / methods
Mice
Myocardium / analysis
Rats
T-Lymphocytes / analysis
Zinc / analysis
Chemical
Reg. No./Substance:
0/Aminoquinolines; 0/Aniline Compounds; 0/Fluorescent Dyes; 7440-66-6/Zinc; 7440-70-2/Calcium; 7782-41-4/Fluorine; 86277-62-5/8-(bis(ethoxycarbonylmethyl)amino)-6-methoxy-2-(bis(ethoxycarbonylmethyl)amino)styrylquinoline

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


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