J Korean Neuropsychiatr Assoc.  2010 Mar;49(2):241-247.

The Regulation of ERK, GSK3beta and AKT after Acute Ethanol Exposure and Withdrawal in SH-SY5Y Human Neuroblastoma Cell-line

• Roh J
• Cui FJ
• Kang UG

Abstract

OBJECTIVES
This study aimed to demonstrate the activities and phosphorylation changes induced by acute ethanol treatment and withdrawal conditions in the intracellular signal transduction molecules [such as extracellular signal-regulated kinase (ERK), glycogen synthase kinase 3beta (GSK3beta), and Akt] of the SH-SY5Y neuroblastoma cell line.
METHODS
The acute treatment exposed SH-SY5Y cells to 100 mM ethanol, and we took samples 30 minutes, 60 minutes, and 24 hours after initiating this treatment. After 24 hours' continuous ethanol treatment, we initiated ethanol withdrawal, taking samples at 30 minutes and 60 minutes. We assayed the kinase phosphorylations via an immunoblot analysis using phosphorspecific antibodies, quantified by optical densitometry.
RESULTS
Ethanol treatment induced a transient increase in phosphorylation of GSK3beta and Akt at 30 minutes but failed to change the phosphorylation level of ERK. Ethanol withdrawal induced a transient ERK phosphorylation increase at 30 minutes, but it had no effect on the phosphorylation of GSK3beta or Akt.
CONCLUSION
The results indicate that the ethanol-induced cellular response includes the ERK, GSK3beta, and Akt systems. In particular, the ERK pathway may play a role in the acute withdrawal response. This also suggests that a relatively short exposure to ethanol, such as the 24-hour exposure in this study, can induce functional adaptation within a cell.

Keyword

Ethanol; Treatment; Withdrawal; ERK; GSK3beta; AKT; Phosphorylation

MeSH Terms

Antibodies
Cell Line
Densitometry
Ethanol
Glycogen Synthase Kinase 3
Glycogen Synthase Kinases
Humans
MAP Kinase Signaling System
Neuroblastoma
Phosphorylation
Phosphotransferases
Signal Transduction
Antibodies
Ethanol
Glycogen Synthase Kinase 3
Glycogen Synthase Kinases
Phosphotransferases