title: Nuclear clusterin is associated with neuronal apoptosis in the developing rat brain upon ethanol exposure.
abstract: BACKGROUND
  Fetal alcohol spectrum disorder (FASD) is often accompanied by reduced brain volumes, reflecting brain cell death induced by ethanol, but the molecular mechanisms were less elucidated. This study was set up to investigate whether clusterin (Clu) was involved in neuronal cell death in developing rats.
  
  
  METHODS
  Seven-day-old rats were subcutaneously injected with 20% ethanol in normal saline at 3 g/kg twice. The upregulation of Clu and cell death was detected by immunohistochemistry, immunofluorescence microscopy, and/or Western blotting. Protein-protein interaction was detected by immunoprecipitation and immunoblotting. To identify the isoform interacting with Bcl-XL, HT22 mouse hippocampal cells were transfected with nuclear Clu(nClu)- or secretory Clu-expressing vector, and confocal microscopy was performed. Clu transcripts were knocked down in primary cortical cells using siRNA.
  
  
  RESULTS
  We found that Clu increased in the cerebral cortex following acute ethanol treatment. The Clu upregulation was related to increased cell death, which was assessed by activated caspase-3 and TUNEL staining. The upregulated Clu was a nuclear isoform that was nuclear translocated upon ethanol exposure and interacted with Bcl-XL, mediating apoptosis.
  
  
  CONCLUSIONS
  This study shows that nClu plays a pro-apoptotic role in ethanol-induced cell death in the developing brain, providing new insights for development of FASD.
authors.affiliation: Department of Anatomy and Neurobiology, Health Science Institute, Medical Research Center for Neural Dysfunction, School of Medicine, Gyeongsang National University, Jinju, Gyeongnam, South Korea.
authors.forename: Nayoung
authors.initials: N
authors.lastname: Kim
authors.forename: Jae Y
authors.initials: JY
authors.lastname: Han
authors.forename: Gu S
authors.initials: GS
authors.lastname: Roh
authors.forename: Hyun J
authors.initials: HJ
authors.lastname: Kim
authors.forename: Sang S
authors.initials: SS
authors.lastname: Kang
authors.forename: Gyeong J
authors.initials: GJ
authors.lastname: Cho
authors.forename: Jae Y
authors.initials: JY
authors.lastname: Park
authors.forename: Wan S
authors.initials: WS
authors.lastname: Choi
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D017209:Apoptosis / Q000187:drug effects*; D001921:Brain / Q000187:drug effects / Q000254:growth & development* / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000187:drug effects / Q000378:metabolism; D051152:Clusterin / Q000378:metabolism*; D000431:Ethanol / Q000008:administration & dosage*; D006624:Hippocampus / Q000166:cytology; D051379:Mice; D051381:Rats; D017207:Rats, Sprague-Dawley; D015854:Up-Regulation
chemical_list: C498500:Clu protein, rat; D051152:Clusterin; D000431:Ethanol