Control of basal extracellular adenosine concentration in rat cerebellum.
To re-examine how the basal extracellular concentration of adenosine is regulated in acutely isolated cerebellar slices we have combined electrophysiological and microelectrode biosensor measurements. In almost all cases, synaptic transmission was tonically inhibited by adenosine acting via A1 receptors. By contrast, in most slices, the biosensors did not measure an adenosine tone but did record a spatially non-uniform extracellular tone of the downstream metabolites (inosine and hypoxanthine). Most of the extracellular hypoxanthine arose from the metabolism of inosine by ecto-purine nucleoside phosphorylase (PNP). Adenosine kinase was the major determinant of adenosine levels, as its inhibition increased both adenosine concentration and A1 receptor-mediated synaptic inhibition. Breakdown of adenosine by adenosine deaminase was the major source of the inosine/hypoxanthine tone. However adenosine deaminase played a minor role in determining the level of adenosine at synapses, suggesting a distal location. Blockade of adenosine transport (by NBTI/dipyridamole) had inconsistent effects on basal levels of adenosine and synaptic transmission. Unexpectedly, application of NBTI/dipyridamole prevented the efflux of adenosine resulting from block of adenosine kinase at only a subset of synapses. We conclude that there is spatial variation in the functional expression of NBTI/dipyridamole-sensitive transporters. The increased spatial and temporal resolution of the purine biosensor measurements has revealed the complexity of the control of adenosine and purine tone in the cerebellum.
Wall
Mark J
MJ
Neuroscience Group, Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, UK. mark.wall@warwick.ac.uk
Atterbury
Alison
A
Dale
Nicholas
N
D000241:Adenosine / Q000378:metabolism*; D000243:Adenosine Deaminase / Q000378:metabolism; D000248:Adenosine Kinase / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D015374:Biosensing Techniques / Q000295:instrumentation; D002531:Cerebellum / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D004558:Electric Stimulation; D004594:Electrophysiology / Q000379:methods; D033705:Equilibrative Nucleoside Transport Proteins / Q000378:metabolism; D019706:Excitatory Postsynaptic Potentials; D005110:Extracellular Space / Q000201:enzymology / Q000378:metabolism*; D019271:Hypoxanthine / Q000378:metabolism; D066298:In Vitro Techniques; D007288:Inosine / Q000378:metabolism; D008297:Male; D008839:Microelectrodes; D009433:Neural Inhibition; D033702:Nucleobase Transport Proteins / Q000378:metabolism; D011683:Purine-Nucleoside Phosphorylase / Q000378:metabolism; D011689:Purkinje Cells / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D043682:Receptor, Adenosine A1 / Q000378:metabolism*; D009435:Synaptic Transmission; D013997:Time Factors
D033705:Equilibrative Nucleoside Transport Proteins; D033702:Nucleobase Transport Proteins; D043682:Receptor, Adenosine A1; D019271:Hypoxanthine; D007288:Inosine; D000255:Adenosine Triphosphate; D011683:Purine-Nucleoside Phosphorylase; D000248:Adenosine Kinase; D000243:Adenosine Deaminase; D000241:Adenosine