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Central ghrelin is required for the rewarding properties of drug abuse. We investigated whether alcohol affects ghrelinergic, dopaminergic, and serotoninergic neurons and growth hormone secretagogue receptor 1A (GHS-R1A) levels in the reward system of the brain. Alcohol-naïve C57BL/6 J mice received 2 g/kg ethanol (EtOH) intraperitoneally (i.p.). Plasma ghrelin levels decreased between 1 and 4 h. We investigated the effects of EtOH administration on plasma ghrelin levels in two different animal models at 1, 3, and 10 months of age. Plasma ghrelin levels decreased following the EtOH treatment in 1- and 3-month-old short-term (1-day) alcohol vapor-exposed (STA) mice. In contrast, EtOH administration increased plasma ghrelin levels in 1- and 3-month-old long-term (20-day) alcohol vapor-exposed (LTA) mice. In vivo ghrelin release in the lateral hypothalamus (LH) increased in STA and LTA mice after the i.p. administration of EtOH. EtOH increased in vivo dopamine (DA), but not serotonin (5-HT) release in the LH of STA mice, and increased in vivo DA and 5-HT release in the LH of LTA mice. GHS-R1A mRNA expression and GHS-R1A protein levels in the LH were increased in LTA mice. The number of GHS-R1A-immunoreactive cells was greater in the LH and amygdala of LTA mice. These results support the neurobiological correlation between the development of drinking behavior and activation of ghrelinergic and serotonergic neurons in the LH. The activation of ghrelinergic systems in the amygdala may also induce an increase in 5-HT release in the LH during long-term alcohol intake.Alcohol decreased plasma ghrelin in one- and 3-month old short- term alcohol vapor-exposed (STA) mice.Alcohol-induced plasma ghrelin levels were increased in the long-term alcohol vapor-exposed (LTA) mice.Lateral hypothalamus (LH) alcohol-induced ghrelin releases were increased in both models.Characteristic increases of LH serotonin releases were shown in the LTA following alcohol administration.Positive ghrelin receptor cells and their proteins were increased in the LH and central amygdala of the only LTA.