The effect of temperature on the respiration of cultured neural cells as studied by a novel electron paramagnetic resonance technique.
A new technique to study cellular respiration under steady-state conditions is described. For detection of oxygen concentration the technique utilizes lithium phthalocyanine crystals whose, electron paramagnetic resonance (EPR) spectra are highly sensitive to oxygen. A gas permeable poly(tetrafluoroethylene) (TFE) capillary (i.d. = 0.81 mm; o.d. = 0.86 mm) is used to control the oxygen flux to the sample of cultured neural cells (N1E-115) of 50 microliters volume. The measured oxygen permeability of the TFE capillary exhibits Arrhenius law behavior (Ep = 21.2 kJ/mole) in the temperature range 15-43 degrees C. The observed increase in the respiratory rate of the N1E-115 cells with temperature can be described with an apparent activation energy of Ea = 110 +/- 20 kJ/mol. Cellular depolarization with 35 mM KCl increases the respiratory rate of the cells but gives the same activation energy. The results confirm a hypothesis that respiratory depolarizers affect only the total enzyme concentration and do not change the apparent Ea of the substrate-enzyme decomposition reaction. The theoretical treatment of oxygen diffusion in the system provides guidelines for designing steady-state cellular respiration experiments.
Smirnov
A I
AI
College of Medicine, Urbana, IL 61801.
Norby
S W
SW
Weyhenmeyer
J A
JA
Clarkson
R B
RB
D000818:Animals; D004058:Diffusion; D004578:Electron Spin Resonance Spectroscopy / Q000379:methods*; D007211:Indoles; D008433:Mathematics; D009447:Neuroblastoma / Q000378:metabolism; D009942:Organometallic Compounds; D010100:Oxygen / Q000032:analysis; D010101:Oxygen Consumption; D013696:Temperature; D014407:Tumor Cells, Cultured
D007211:Indoles; D009942:Organometallic Compounds; C081756:lithium phthalocyanine; D010100:Oxygen