title: Antagonism of adenosine receptors by caffeine and caffeine metabolites in equine forebrain tissues.
abstract: OBJECTIVE
  To determine the presence of adenosine receptor subtypes A1 and A2a in equine forebrain tissues and to characterize the interactions of caffeine and its metabolites with adenosine receptors in the CNS of horses.
  
  
  METHODS
  Brain tissue specimens obtained during necropsy from 5 adult male research horses.
  
  
  METHODS
  Membrane-enriched homogenates from cerebral cortex and striatum were evaluated by radioligand binding assays with the A1-selective ligand [3H]DPCPX and the A2a-selective ligand [3H]ZM241385. Functional responses to adenosine receptor agonists and antagonists were determined by a nucleotide exchange assay using [35S]-guanosine 5'-(gamma-thio) triphosphate ([35S]GTPgammaS).
  
  
  RESULTS
  Saturable high affinity [3H]DPCPX binding (A1) sites were detected in cerebral cortex and striatum, whereas high-affinity [3H]ZM241385 binding (A2a) sites were detected only in striatum. Caffeine and related methylxanthines had similar binding affinities at A1 and A2a sites with rank orders of drug binding affinities (theophylline > paraxanthine > or = caffeine >> theobromine) similar to other species. [35S]GTPgammaS exchange revealed that caffeine and its metabolites act as pure adenosine receptor antagonists at concentrations that correspond to A1 and A2a receptor binding affinities.
  
  
  CONCLUSIONS
  Results of our study affirm the presence of guanine nucleotide binding protein linked adenosine receptors (ie, high-affinity A1 and A2a adenosine receptors) in equine forebrain tissues and reveal the antagonistic actions by caffeine and several biologically active caffeine metabolites. Antagonism of adenosine actions in the equine CNS by these stimulants may be responsible for some central actions of methylxanthine drugs, including motor stimulation and enhanced racing performance.
authors.affiliation: Department of Physiological Sciences, College of Veterinary Medicine, University of Florida, Gainesville, FL 32610-0144, USA.
authors.forename: Chi-Chung
authors.initials: CC
authors.lastname: Chou
authors.forename: Thomas W
authors.initials: TW
authors.lastname: Vickroy
mesh_terms: D000818:Animals; D002110:Caffeine / Q000378:metabolism* / Q000494:pharmacology*; D000697:Central Nervous System Stimulants / Q000378:metabolism / Q000494:pharmacology; D002540:Cerebral Cortex / Q000378:metabolism; D006736:Horses; D008297:Male; D017072:Neostriatum / Q000378:metabolism; D016548:Prosencephalon / Q000378:metabolism*; D058915:Purinergic P1 Receptor Antagonists; D018047:Receptors, Purinergic P1 / Q000378:metabolism*; D013805:Theobromine / Q000378:metabolism; D013806:Theophylline / Q000378:metabolism
chemical_list: D000697:Central Nervous System Stimulants; D058915:Purinergic P1 Receptor Antagonists; D018047:Receptors, Purinergic P1; D002110:Caffeine; D013806:Theophylline; D013805:Theobromine; C021183:1,7-dimethylxanthine