title: Afobazole modulates neuronal response to ischemia and acidosis via activation of sigma-1 receptors.
abstract: Afobazole is an anxiolytic medication that has been previously shown to be neuroprotective both in vitro and in vivo. However, the mechanism(s) by which afobazole can enhance neuronal survival remain poorly understood. Experiments were carried out to determine whether afobazole can decrease intracellular calcium overload associated with ischemia and acidosis and whether the effects of afobazole are mediated via interaction of the compound with σ receptors. Fluorometric Ca(2+) imaging was used to resolve how application of afobazole affects intracellular Ca(2+) handling in cortical neurons. Application of afobazole significantly depressed, in a concentration-dependent and reversible manner, the intracellular Ca(2+) overload resulting from in vitro ischemia and acidosis. The IC(50) for afobazole inhibition of ischemia-evoked intracellular Ca(2+) overload was considerably less than that for the inhibition of [Ca(2+)](i) increases induced by acidosis. However, afobazole maximally inhibited only 70% of the ischemia-evoked intracellular Ca(2+) overload but effectively abolished intracellular Ca(2+) increases produced by acidosis. The effects of afobazole on ischemia- and acidosis-induced intracellular Ca(2+) dysregulation were inhibited by preincubating the neurons in the irreversible, pan-selective σ-receptor antagonist, metaphit. Moreover, the effects of afobazole on intracellular Ca(2+) increases triggered by acidosis and ischemia were blocked by the selective σ-1-receptor antagonists, BD 1063 and BD 1047, respectively. Experiments examining the effects of afobazole on neuronal survival in response to ischemia showed that afobazole was neuroprotective. Taken together, these data suggest that afobazole regulates intracellular Ca(2+) overload during ischemia and acidosis via activation of σ-1 receptors. This mechanism is probably responsible for afobazole-mediated neuroprotection.
authors.affiliation: Department of Molecular Pharmacology and Physiology, University of South Florida College of Medicine, Tampa, Florida 33612-4799, USA. jcuevas@health.usf.edu
authors.forename: Javier
authors.initials: J
authors.lastname: Cuevas
authors.forename: Adam
authors.initials: A
authors.lastname: Behensky
authors.forename: Wei
authors.initials: W
authors.lastname: Deng
authors.forename: Christopher
authors.initials: C
authors.lastname: Katnik
mesh_terms: D000138:Acidosis / Q000378:metabolism* / Q000473:pathology; D000818:Animals; D014151:Anti-Anxiety Agents / Q000494:pharmacology*; D001562:Benzimidazoles / Q000494:pharmacology*; D002545:Brain Ischemia / Q000378:metabolism* / Q000473:pathology; D002118:Calcium / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D002540:Cerebral Cortex / Q000166:cytology / Q000187:drug effects; D005029:Ethylenediamines / Q000494:pharmacology; D005260:Female; D006146:Guanidines / Q000494:pharmacology; D007202:Indicators and Reagents; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008564:Membrane Potentials / Q000187:drug effects; D009025:Morpholines / Q000494:pharmacology*; D009294:Narcotics / Q000494:pharmacology; D009474:Neurons / Q000378:metabolism* / Q000473:pathology; D018408:Patch-Clamp Techniques; D010423:Pentazocine / Q000494:pharmacology; D010622:Phencyclidine / Q000031:analogs & derivatives / Q000494:pharmacology; D010879:Piperazines / Q000494:pharmacology; D011247:Pregnancy; D051381:Rats; D017480:Receptors, sigma / Q000819:agonists* / Q000037:antagonists & inhibitors; D000097605:Sigma-1 Receptor
chemical_list: D014151:Anti-Anxiety Agents; D001562:Benzimidazoles; D002118:Calcium; D005029:Ethylenediamines; D006146:Guanidines; D007202:Indicators and Reagents; D007770:L-Lactate Dehydrogenase; D009025:Morpholines; D009294:Narcotics; D010423:Pentazocine; D010622:Phencyclidine; D010879:Piperazines; D017480:Receptors, sigma; D000097605:Sigma-1 Receptor; C093337:1-(2-(3,4-dichlorophenyl)ethyl)-4-methylpiperazine; C410924:2-((2-morpholino)ethylthio)-5-ethoxybenzimidazole; C097963:N-(2-(3,4-Dichlorphenyl)ethyl)-N,N',N'-trimethyl-1,2-ethandiamin; C044705:metaphit; C050232:1,3-ditolylguanidine