title: Haloperidol and clozapine block formation of autophagolysosomes in rat primary neurons.
abstract: Early intervention and maintenance treatment for schizophrenia patients may prolong the duration of exposure to antipsychotic agents; however, there have been few studies on the neurotoxicity of these agents. Here, we investigated the effects of antipsychotics on cell viability and autophagy in rat primary neurons. Cultured cortical neurons obtained from rat embryos were treated with various concentrations of haloperidol and clozapine, and the neuronal toxicity was assessed by measuring lactate dehydrogenase (LDH) activity and 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay. Autophagosomes were quantitated by measuring the level of microtubule-associated protein 1A/1B-light chain 3 (LC3-II) by Western blot and immunofluorescence staining. Autophagic flux was assayed using bafilomycin A1 and GFP-mCherry-LC3 transfection. Haloperidol and clozapine decreased the viability of neurons in vitro in a concentration- and time-dependent manner. We also observed increased accumulation of autophagosomes after antipsychotic treatment. Using bafilomycin A1 and GFP-mCherry-LC3 transfection, we discovered that haloperidol and clozapine inhibited autophagosome turnover resulting in a dysfunctional autophagic process, including impaired lysosomal fusion. Together, these results suggest that haloperidol and clozapine negatively affect neuronal viability, possibly by blocking autophagolysosome formation.
authors.affiliation: Department of Psychiatry, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea.
authors.forename: J
authors.initials: J
authors.lastname: Park
authors.forename: S
authors.initials: S
authors.lastname: Chung
authors.forename: H
authors.initials: H
authors.lastname: An
authors.forename: J
authors.initials: J
authors.lastname: Kim
authors.forename: J
authors.initials: J
authors.lastname: Seo
authors.forename: D H
authors.initials: DH
authors.lastname: Kim
authors.forename: S Y
authors.initials: SY
authors.lastname: Yoon
mesh_terms: D000818:Animals; D014150:Antipsychotic Agents / Q000633:toxicity*; D001343:Autophagy / Q000187:drug effects*; D015153:Blotting, Western; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003024:Clozapine / Q000633:toxicity*; D005455:Fluorescent Antibody Technique; D006220:Haloperidol / Q000633:toxicity*; D009474:Neurons / Q000187:drug effects*; D010588:Phagosomes / Q000187:drug effects*; D051381:Rats
chemical_list: D014150:Antipsychotic Agents; D003024:Clozapine; D006220:Haloperidol