Metabolic regulation of ATP breakdown and of adenosine production in rat brain extracts.
ATP concentration is dramatically affected in ischemic injury. From previous studies on ATP mediated purine and pyrimidine salvage in CNS, we observed that when "post-mitochondrial" extracts of rat brain were incubated with ATP at 3.6 mM, a normoxic concentration, formation of IMP always preceded that of adenosine, a well known neuroactive nucleoside and a homeostatic cellular modulator. This observation prompted us to undertake a study aimed at assessing the precise pathways and kinetics of ATP breakdown, a process considered to be the major source of adenosine in rat brain. The results obtained using post-mitochondrial extracts strongly suggest that the breakdown of intracellular ATP at normoxic concentration follows almost exclusively the pathway ATP<=>ADP<=>AMP --> IMP --> inosine<=>hypoxanthine, with little, if any, intracellular adenosine production. At low ischemic concentration, intracellular ATP breakdown follows the pathway ATP<=>ADP<=>AMP --> adenosine --> inosine<=>hypoxanthine with little IMP formation. At the same time, extracellular ATP, whose concentration is known to be enhanced during ischemia, is actively broken down to adenosine through the pathway ATP --> ADP --> AMP --> adenosine, catalysed by the well characterized ecto-enzyme cascade system. Moreover, we show that during intracellular GTP catabolism, xanthosine, in addition to guanosine, is generated through the so called "ribose 1-phosphate recycling for nucleoside interconversion". These results considerably extend our knowledge on the long debated question of the extra or intracellular origin of adenosine in CNS, suggesting that at least in normoxic conditions, intracellular adenosine is of extracellular origin.
Barsotti
Catia
C
Department of Physiology and Biochemistry, University of Pisa, Via San Zeno 51, 56127 Pisa, Italy. cbarsotti@dfb.unipi.it
Ipata
Piero L
PL
D000241:Adenosine / Q000378:metabolism*; D000249:Adenosine Monophosphate / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D001921:Brain / Q000201:enzymology*; D006151:Guanosine / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000378:metabolism; D019271:Hypoxanthine / Q000378:metabolism; D007288:Inosine / Q000378:metabolism; D007291:Inosine Monophosphate / Q000378:metabolism*; D051381:Rats
D006151:Guanosine; D007291:Inosine Monophosphate; D019271:Hypoxanthine; D000249:Adenosine Monophosphate; D007288:Inosine; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D000241:Adenosine