Extracellular adenosine concentrations during in vitro ischaemia in rat hippocampal slices

Br J Pharmacol. 1999 Jun;127(3):729-39. doi: 10.1038/sj.bjp.0702591.

Abstract

1. The application of an ischaemic insult in hippocampal slices results in the depression of synaptic transmission, mainly attributed to the activation of A1 adenosine receptors by adenosine released in the extracellular space. 2. To estimate the concentration of endogenous adenosine acting at the receptor level during an ischaemic episode, we recorded field e.p.s.ps (fe.p.s.ps) from hippocampal slices, and evaluated the ability of the selective A1 receptor antagonist, 8-cyclopentyl-1,3-dipropylxanthine (DPCPX), to reverse the fe.p.s.p. depression induced by in vitro ischaemia. A relationship between the IC50 of an antagonist and the endogenous concentration of a neurotransmitter has been used for pharmacological analysis. 3. The complete and reversible depression of fe.p.s.p. in the CA1 region induced by 5 min ischaemia was decreased in the presence of DPCPX (50-500 nM). 8-Phenyltheophylline (10 microM) abolished the depression of fe.p.s.ps during the ischaemic period, while a small (peak effect 12 +/- 4%) decrease in fe.p.s.ps was observed during the initial phase of reperfusion. 4. In the time-interval of maximal depression of fe.p.s.ps., IC50 and adenosine concentration changed as function of time with a good degree of correlation. The maximal value of adenosine concentration was 30 microM. 5. Our data provide an estimation of the adenosine concentration reached at the receptor level during an ischaemic episode, with a higher time discrimination (15 s) than that achieved with any biochemical approach. This estimation may be useful in order to establish appropriate concentrations of purinergic compounds to be tested for their pharmacological effects during an ischaemic episode.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine / metabolism*
  • Adenosine / physiology
  • Animals
  • Brain Ischemia / metabolism*
  • Brain Ischemia / physiopathology
  • Extracellular Space / metabolism*
  • Hippocampus / blood supply*
  • In Vitro Techniques
  • Kinetics
  • Male
  • Oxygen / metabolism
  • Purinergic P1 Receptor Antagonists
  • Rats
  • Rats, Wistar
  • Synaptic Transmission / drug effects
  • Synaptic Transmission / physiology
  • Theophylline / analogs & derivatives
  • Theophylline / pharmacology
  • Xanthines / pharmacology

Substances

  • Purinergic P1 Receptor Antagonists
  • Xanthines
  • 1,3-dipropyl-8-cyclopentylxanthine
  • Theophylline
  • 8-phenyltheophylline
  • Adenosine
  • Oxygen