Increase in the ADP/ATP exchange in rat liver mitochondria irradiated in vitro by helium-neon laser

Abstract

To gain some insight into the mechanism of cell photostimulation by laser light, measurements were made of the rate of ADP/ATP exchange in mitochondria irradiated with the low power continuous wave Helium Neon laser (energy dose 5 Joules/cm2). To do this a method has been developed to continuously monitor ATP efflux from phosphorylating mitochondria caused by externally added ADP, by photometrically following the NADP+ reduction which occurs in the presence of glucose, hexokinase, glucose-6-phosphate dehydrogenase and effluxed ATP. The NADP+ reduction rate shows hyperbolic dependence on ADP concentration (Km and Vmax values 8.5 +/- 0.87 microM and 20.7 +/- 0.49 nmoles NADP+ reduced/min x mg mitochondrial protein, respectively), and proves to measure the activity of the ADP/ATP translocator as shown by inhibition experiments using atracyloside, powerful inhibitor of this carrier. Irradiation was found to enhance the rate of ADP/ATP antiport, with externally added ADP ranging between 5 and 100 microM. As a result of experiments carried out with mitochondria loaded with either ATP or ADP, the increase in the activity of the ADP/ATP translocator is here proposed to depend on the increase in the electrochemical proton gradient which occurs owing to irradiation of mitochondria.