Extracellular levels of adenosine and its metabolites in the striatum of awake rats: inhibition of uptake and metabolism.
The level of purines in the striatum of awake, freely moving rats was studied using microdialysis. The calculated extracellular concentration of adenosine and its metabolites inosine and hypoxanthine was very high immediately after implantation of the dialysis probe but decreased within 24 h to a level which remained stable for two days. Using in vitro calibration to determine the relative recovery of the dialysis probes we estimated resting levels in the striatal extracellular space to be 40, 110 and 580 nM, respectively. Inhibition of adenosine deaminase by deoxycoformycin produced a significant 1.4-fold increase in extracellular adenosine levels and a fall in inosine and hypoxanthine. A combination of three uptake blockers (dipyridamole, lidoflazine and nitrobenzylthioinosine), caused a 4.5-fold increase in extracellular adenosine levels without any change in inosine or hypoxanthine levels. After uptake inhibition deoxycoformycin did not have any significant effect. The present results show that the microdialysis technique can be used to determine levels of purines in the extracellular fluid of defined brain regions in awake animals. The high levels recorded during the first several hours after implantation may be artefactually high and reflect trauma. The results also show that adenosine levels can be altered in vivo by inhibitors of adenosine transport and adenosine deaminase. The present results indicate that the physiological adenosine level in striatal extracellular space is in the range 40-460 nM.
Ballarín
M
M
Department of Biochemistry, Faculty of Medicine, University of Barcelona, Spain.
Fredholm
B B
BB
Ambrosio
S
S
Mahy
N
N
D000241:Adenosine / Q000378:metabolism*; D058892:Adenosine Deaminase Inhibitors; D000818:Animals; D001693:Biological Transport, Active / Q000187:drug effects; D003342:Corpus Striatum / Q000187:drug effects / Q000378:metabolism*; D004176:Dipyridamole / Q000494:pharmacology; D005110:Extracellular Space / Q000378:metabolism; D019271:Hypoxanthine; D007042:Hypoxanthines / Q000378:metabolism; D007288:Inosine / Q000378:metabolism; D008297:Male; D015649:Pentostatin / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains
D058892:Adenosine Deaminase Inhibitors; D007042:Hypoxanthines; D019271:Hypoxanthine; D015649:Pentostatin; D007288:Inosine; D004176:Dipyridamole; D000241:Adenosine