Beta-oxidation of docosahexaenoic acid and tetracosahexaenoic acid in isolated rat hepatocytes and human fibroblasts

2001 ◽  
Vol 29 (1) ◽  
pp. A23-A23
Author(s):  
K. De Nys ◽  
E. Meyhi ◽  
S. Asselberghs ◽  
G. Mannaerts ◽  
P. Van Veldhoven
Keyword(s):  
Docosahexaenoic Acid ◽  
Human Fibroblasts ◽  
Rat Hepatocytes ◽  
Beta Oxidation ◽  
Isolated Rat Hepatocytes ◽  
Tetracosahexaenoic Acid ◽  
Isolated Rat

scholarly journals Continuous bioluminescent monitoring of cytoplasmic ATP in single isolated rat hepatocytes during metabolic poisoning

1993 ◽  
Vol 295 (1) ◽  
pp. 165-170 ◽  
Author(s):  
A Koop ◽  
P H Cobbold
Keyword(s):  
Time Course ◽  
Rat Hepatocytes ◽  
Firefly Luciferase ◽  
Beta Oxidation ◽  
Isolated Rat Hepatocytes ◽  
Chemical Hypoxia ◽  
Luminescent Signal ◽  
Bioluminescent Monitoring ◽  
Cell To Cell Variability ◽  
Isolated Rat

We have devised a technique for monitoring cytoplasmic ATP continuously in single hepatocytes. Single isolated rat hepatocytes were injected with the ATP-dependent luminescent protein firefly luciferase, and then superfused with 45 microM luciferin in air-equilibrated medium. Signals of approx. 10-200 photoelectron counts per second could be recorded from individual healthy cells for up to 3 h. The response of the luminescent signal to chemical hypoxia (2-5 mM CN- and 5-10 mM 2-deoxyglucose) was monitored. We found a great cell-to-cell variability in the time course of the ATP decline in response to CN-, 2-deoxyglucose or to their combination; the time for the signal to fall to 10% of the original (corresponding to approx. 100 microM ATP) ranged from approx. 20 to 75 min. This resistance of the cytoplasmic ATP concentration to depletion after blockade of oxidative phosphorylation and glycolysis could be abolished by pretreatment of the cells with etomoxir, which blocks mitochondrial beta-oxidation. Etomoxir alone had no effect on the luciferase signal, but etomoxir-pre-treated cells showed a prompt fall in the luciferase signal starting within 1-2 min of application of cyanide and 2-deoxyglucose and falling to 10% of the original signal in approx. 6-10 min. The technique allows cytoplasmic ATP changes to be monitored in single hepatocytes at concentrations of 1 mM or lower, but more precise calibration of the signal will require correction for the effects of cytoplasmic pH changes.

scholarly journals Lysosomal triacylglycerol lipase and lipolysis in isolated rat hepatocytes.

1979 ◽  
Vol 254 (18) ◽  
pp. 8841-8846
Author(s):  
L.J. Debeer ◽  
J. Thomas ◽  
P.J. De Schepper ◽  
G.P. Mannaerts
Keyword(s):  
Rat Hepatocytes ◽  
Isolated Rat Hepatocytes ◽  
Triacylglycerol Lipase ◽  
Isolated Rat
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