title: Alcohol disrupts endoplasmic reticulum function and protein secretion in hepatocytes.
abstract: BACKGROUND
  Many alcoholic patients have serum protein deficiency that contributes to their systemic problems. The unfolded protein response (UPR) is induced in response to disequilibrium in the protein folding capability of the endoplasmic reticulum (ER) and is implicated in hepatocyte lipid accumulation and apoptosis, which are associated with alcoholic liver disease (ALD). We investigated whether alcohol affects ER structure, function, and UPR activation in hepatocytes in vitro and in vivo.
  
  
  METHODS
  HepG2 cells expressing human cytochrome P450 2E1 and mouse alcohol dehydrogenase (VL-17A) were treated for up to 48 hours with 50 and 100 mM ethanol. Zebrafish larvae at 4 days postfertilization were exposed to 350 mM ethanol for 32 hours. ER morphology was visualized by fluorescence in cells and transmission electron microscopy in zebrafish. UPR target gene activation was assessed using quantitative PCR, in situ hybridization, and Western blotting. Mobility of the major ER chaperone, BIP, was monitored in cells by fluorescence recovery after photobleaching (FRAP).
  
  
  RESULTS
  VL-17A cells metabolized alcohol yet only had slight activation of some UPR target genes following ethanol treatment. However, ER fragmentation, crowding, and accumulation of unfolded proteins as detected by immunofluorescence and FRAP demonstrate that alcohol induced some ER dysfunction despite the lack of UPR activation. Zebrafish treated with alcohol, however, showed modest ER dilation, and several UPR targets were significantly induced.
  
  
  CONCLUSIONS
  Ethanol metabolism directly impairs ER structure and function in hepatocytes. Zebrafish are a novel in vivo system for studying ALD.
authors.affiliation: Division of Liver Diseases, Department of Medicine, Mount Sinai School of Medicine, New York, New York, USA.
authors.forename: Deanna L
authors.initials: DL
authors.lastname: Howarth
authors.forename: Ana M
authors.initials: AM
authors.lastname: Vacaru
authors.forename: Orkhontuya
authors.initials: O
authors.lastname: Tsedensodnom
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Mormone
authors.forename: Natalia
authors.initials: N
authors.lastname: Nieto
authors.forename: Lindsey M
authors.initials: LM
authors.lastname: Costantini
authors.forename: Erik L
authors.initials: EL
authors.lastname: Snapp
authors.forename: Kirsten C
authors.initials: KC
authors.lastname: Sadler
mesh_terms: D000818:Animals; D004721:Endoplasmic Reticulum / Q000187:drug effects* / Q000648:ultrastructure; D000431:Ethanol / Q000633:toxicity*; D056945:Hep G2 Cells; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism*; D006801:Humans; D051379:Mice; D056811:Unfolded Protein Response / Q000187:drug effects*; D015027:Zebrafish
chemical_list: D000431:Ethanol