Figure 1.
Acute JMV2959 treatment decreases alcohol intake in rats.
(A) The acute effects of the GHS-R1A antagonist JMV2959 (1 or 3 mg/kg) administration on alcohol intake was evaluated at 1 hour time interval after both two and five months of voluntary alcohol consumption. (B) The acute effects of the GHS-R1A antagonist JMV2959 (1 or 3 mg/kg) administration on alcohol intake was evaluated at 4 hour time interval after both two and five months of voluntary alcohol consumption. JMV2959 significantly decreases voluntary alcohol consumption compared to vehicle and the effect was more pronounced after five compared to two months of alcohol consumption. All values represent mean alcohol intake (g/kg) ± SEM (n = 19, *P<0.05, **P<0.01, ***P<0.001 compared to corresponding vehicle or as indicated).
Figure 2.
Acute JMV2959 treatment increases water intake in rats.
(A) Acute administration of the GHS-R1A antagonist JMV2959 (3 mg/kg) significantly increased water consumption compared to vehicle at 1 hour time interval in rats voluntarily consuming alcohol for two and five months before the treatment. (B) Acute administration of the GHS-R1A antagonist JMV2959 (3 mg/kg) significantly increased water consumption compared to vehicle at 4 hour time interval in rats voluntarily consuming alcohol for two and five months before the treatment. All values represent mean water intake (ml) ± SEM (n = 19, ***P<0.001 compared to corresponding vehicle).
Figure 3.
Acute JMV2959 treatment decreases alcohol preference in rats.
(A) Acute administration of the GHS-R1A antagonist JMV2959 (1 or 3 mg/kg) decreases the alcohol preference compared to vehicle at 1 hour time interval in rats voluntarily consuming alcohol for two and five months before the treatment. Acute administration of the GHS-R1A antagonist JMV2959 (1 or 3 mg/kg) decreases the alcohol preference compared to vehicle at 4 hour time interval in rats voluntarily consuming alcohol for two and five months before the treatment. All values represent mean alcohol preference (%) ± SEM (n = 19, *P<0.05, **P<0.01, ***P<0.001 compared to corresponding vehicle or as indicated).
Figure 4.
Repeated JMV2959 treatment decreases alcohol intake in rats.
(A) Repeated administration of the GHS-R1A antagonist JMV2959 (3 mg/kg) significantly decreased voluntary alcohol consumption compared to vehicle at 1 hour time interval in rats voluntarily consuming alcohol for eight months before the treatment. (B) Repeated administration of the GHS-R1A antagonist JMV2959 (3 mg/kg) significantly decreased voluntary alcohol consumption compared to vehicle at 24 hour time interval in rats voluntarily consuming alcohol for eight months before the treatment. No tolerance to treatment was observed and no rebound drinking following discontinuation of treatment was observed. All values represent mean alcohol intake (g/kg) ± SEM (n = 19, *P<0.05, **P<0.01, ***P<0.001 compared to vehicle on the corresponding treatment day).
Figure 5.
Repeated JMV2959 treatment increases water intake in rats.
(A) Repeated administration of the GHS-R1A antagonist JMV2959 (3 mg/kg) significantly increased water intake compared to vehicle at the 1 hour time interval in rats voluntarily consuming alcohol for eight months before the treatment. (B) Repeated administration of the GHS-R1A antagonist JMV2959 (3 mg/kg) significantly decreased alcohol preference compared to vehicle at the 1 hour time interval in rats voluntarily consuming alcohol for eight months before the treatment. All values represent mean ± SEM (n = 19, *P<0.05, **P<0.01, ***P<0.001 compared to vehicle on the corresponding treatment day).
Figure 6.
Acute JMV2959 treatment prevents the alcohol deprivation effect in rats.
Following seven weeks of alcohol consumption (baseline, white bars) the rats were exposed to ten days of alcohol abstinence. Before alcohol was reintroduced (black bars) the rats were treated with wither JMV2959 or vehicle. An alcohol deprivation effect was observed in vehicle treated rats but not in rats treated with the GHS-R1A antagonist, JMV2959. Data are presented as mean alcohol intake (g/kg/1 hr) ± SEM. (n = 15 in each group, *P<0.05 compared to corresponding baseline).
Figure 7.
Scatter plot of Ghsr expression in the VTA.
The present figure shows a significant negative correlation between the expression of Ghsr in the VTA (data presented as arbitrary units 2−ΔΔCT where the mean ΔCT of the group of low alcohol consuming rats was set as the calibrator) and mean alcohol consumption (g/kg/24 hrs) in rats that have voluntarily been drinking alcohol for ten months.
Table 1.
Ghsr mRNA expression in reward related areas of low (2.3±0.2 g/kg/24 hr) and high (4.5±0.2 g/kg/24 hr alcohol-consuming rats after approximately ten months of voluntary alcohol consumption.
Table 2.
Data expressed as methylation degree (%±SD), both overall for the CpG island and at the individual CpG sites, of the Ghsr in ventral tegmental area of low (2.3±0.2 g/kg/24 hr) and high (4.5±0.2 g/kg/24 hr) alcohol-consuming rats after approximately ten months of voluntary alcohol consumption.