Abstract
Introduction . Mechanistic target of rapamycin complex 2 (mTORC2) mediates binge intake of ethanol in rodents and thereby facilitates behavioural sensitization towards the effects of ethanol. Generally, mTORC2 acts as an upstream kinase by phosphorylating other downstream kinases through its sub-components, mTOR and sin1. Amygdala is the brain region that regulates emotion and has been associated with various behavioural changes in alcohol use disorder (AUD). To date, little is known about the effects of ethanol on mTORC2, especially in amygdala. Therefore, this study was undertaken to evaluate the changes in protein expression of mTOR and sin1 in the amygdala of male Wistar rats. Methods . Seven-weeks-old male Wistar rats (n=18) were allocated into three different groups: control, acute, and chronic ethanol. The control group was fed solely with Modified Liquid Diet (MLD) (without ethanol) which included low fat milk powder, maltodextrin and sucrose for 27 days. The acute group was given control MLD for 27 days and ethanol (2.5 g/kg, 20% v/v in saline) was administered once, on day 28 via intra peritoneal route. For chronic group, MLD without ethanol was given for first 7 days and followed by gradual introduction of ethanol at 2.4% (3 days), 4.8% (3 days) and 7.2% (14 days). Parameters such as body weight (g), MLD intake (ml), ethanol intake (g/kg) and serum ethanol level (mg/dl) were measured. At the end, amygdala was dissected bilaterally and the target protein expressions were analysed using Western Blot. Results . The relative comparison of chronic group with control showed significant increase in the protein expression of mTOR and sin1. However, no relative difference was noticed for acute ethanol. Conclusion . Collectively, our data indicate changes in the protein expression of mTORC2 components following chronic ethanol intake, suggesting their probable roles in AUD at chronic stage rather than acute in the amygdala.
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