Alcohol Disrupts Endoplasmic Reticulum Function and Protein Secretion in Hepatocytes
- Authors
- Howarth, D.L., Vacaru, A.M., Tsedensodnom, O., Mormone, E., Nieto, N., Costantini, L.M., Snapp, E.L., and Sadler, K.C.
- ID
- ZDB-PUB-110803-14
- Date
- 2012
- Source
- Alcoholism, clinical and experimental research 36(1): 14-23 (Journal)
- Registered Authors
- Howarth, Deanna, Sadler Edepli, Kirsten C., Tsedensodnom, OonoO, Vacaru, Ana
- Keywords
- ethanol, liver, unfolded protein response, alcoholic liver disease, zebrafish, fuorescence recovery after photobleaching
- MeSH Terms
-
- Animals
- Endoplasmic Reticulum/drug effects*
- Endoplasmic Reticulum/ultrastructure
- Ethanol/toxicity*
- Hep G2 Cells
- Hepatocytes/drug effects*
- Humans
- Mice
- Unfolded Protein Response/drug effects*
- Zebrafish
- PubMed
- 21790674 Full text @ Alcoholism Clin. Exp. Res.
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.