title: Neurotoxicity in organotypic hippocampal slices mediated by adenosine analogues and nitric oxide.
abstract: Adenosine (ADO) and nitric oxide (NO) have been implicated in a variety of neurophysiological actions, including induction of long-term potentiation, regulation of cerebral blood flow, and neurotoxicity/neuroprotection. ADO has been shown to promote NO release from astrocytes by a direct effect on A1 and A2 receptors, thus providing a link between actions of NO and adenosine in the brain. However, while adenosine acts as an endogenous neuroprotectant, NO is believed to be the effector of glutamate neurotoxicity. To resolve this apparent paradox, we have further investigated the effects of adenosine and NO on neuronal viability in cultured organotypic hippocampal slices exposed to sub-lethal (20') in vitro ischemia. Up to a concentration of 500 microM ADO did not cause toxicity while exposures to 100 microM of the stable ADO analogue chloroadenosine (CADO) caused widespread neuronal damage when paired to anoxia/hypoglycemia. CADO effects were significantly prevented by the ADO receptor antagonist theophylline and blockade of NO production by L-NA (100 microM). Moreover, CADO effects were mimicked by the NO donor SIN-1 (100 microM). Application of 100 microM ADO following blockade of adenosine deaminase (with 10 microM EHNA) replicated the effects of CADO. CADO, ADO + EHNA but not ADO alone caused a prolonged and sustained release of nitric oxide as measured by direct amperometric detection. We conclude that at high concentrations and/or following blockade of its enzymatic catabolism, ADO may cause neurotoxicity by triggering NO release from astrocytes. These results demonstrate for the first time that activation of pathways other than those involving neuronal glutamate receptors can trigger NO-mediated neuronal cell death in the hippocampus.
authors.affiliation: Department of Neurological Surgery, University of Washington, Harborview Medical Center, Seattle 98104, USA.
authors.forename: A
authors.initials: A
authors.lastname: Barth
authors.forename: D W
authors.initials: DW
authors.lastname: Newell
authors.forename: L B
authors.initials: LB
authors.lastname: Nguyen
authors.forename: H R
authors.initials: HR
authors.lastname: Winn
authors.forename: R
authors.initials: R
authors.lastname: Wender
authors.forename: J R
authors.initials: JR
authors.lastname: Meno
authors.forename: D
authors.initials: D
authors.lastname: Janigro
mesh_terms: D015762:2-Chloroadenosine / Q000494:pharmacology; D000225:Adenine / Q000031:analogs & derivatives / Q000494:pharmacology; D000241:Adenosine / Q000031:analogs & derivatives* / Q000494:pharmacology; D058892:Adenosine Deaminase Inhibitors; D000818:Animals; D000831:Animals, Newborn; D002545:Brain Ischemia / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006624:Hippocampus / Q000098:blood supply / Q000378:metabolism*; D009474:Neurons / Q000737:chemistry / Q000166:cytology / Q000201:enzymology; D009498:Neurotoxins / Q000494:pharmacology; D009569:Nitric Oxide / Q000378:metabolism*; D009924:Organ Culture Techniques; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D018047:Receptors, Purinergic P1 / Q000378:metabolism; D013805:Theobromine / Q000031:analogs & derivatives / Q000494:pharmacology; D013806:Theophylline / Q000494:pharmacology; D014970:Xanthines / Q000494:pharmacology
chemical_list: D058892:Adenosine Deaminase Inhibitors; D004791:Enzyme Inhibitors; D009498:Neurotoxins; D010726:Phosphodiesterase Inhibitors; D018047:Receptors, Purinergic P1; D014970:Xanthines; D015762:2-Chloroadenosine; D009569:Nitric Oxide; C010192:9-(2-hydroxy-3-nonyl)adenine; C057837:3,7-dimethyl-1-propargylxanthine; C051360:1,3-dipropyl-8-cyclopentylxanthine; D013806:Theophylline; D000225:Adenine; D000241:Adenosine; D013805:Theobromine