Effects of Melanocortin Receptor Activation and Blockade on Ethanol Intake: A Possible Role for the Melanocortin-4 Receptor

THE MELANOCORTIN (MC) system is composed of peptides that are cleaved from the polypeptide precursor proopiomelanocortin (POMC). Central MC peptides are produced by neurons within the hypothalamic arcuate nucleus and the medulla (Dores et al., 1986; Jacobowitz and O’Donohue, 1978; O’Donohue and Dorsa, 1982). These peptides include adrenocorticotropic hormone, α-melanocyte–stimulating hormone (α-MSH), β-MSH, and γ-MSH (Hadley and Haskell-Luevano, 1999). Genetic and pharmacological evidence reveals that MC receptor (MCR) signaling is involved in grooming behavior (Gispen et al., 1975), antipyretic (Murphy et al., 1983) and anti-inflammatory (Macaluso et al., 1994) responses, learning (Zhao et al., 1995), reproductive function (Schioth and Watanobe, 2002), and regulation of appetite and energy homeostasis (Chen et al., 2000; Fan et al., 1997; Huszar et al., 1997; Marsh et al., 1999; Schwartz and Wisse, 2000). There are several observations that suggest that the MC system is a prime candidate for regulating drug-seeking behavior. MCRs are expressed in brain regions thought to mediate drug self-administration, including the nucleus accumbens (NAcc), the hypothalamus, and the ventral tegmental area (VTA) (Alvaro et al., 1996; Griffon et al., 1994; Mountjoy et al., 1994; Roselli-Rehfuss et al., 1993). Interestingly, α-MSH administered into the VTA increases dopamine and 3,4-dihydroxyphenylacetic acid (DOPAC) levels in the NAcc (Lindblom et al., 2001), and chronic intracerebroventricular (ICV) infusion of the nonselective MCR agonist melanotan II (MTII) to rats increases dopamine D1 receptor binding in the NAcc and dopamine D2 receptor binding in the VTA, suggesting that MTII alters dopamine signaling in these regions (Lindblom et al., 2002a). Chronic treatment of a high dose of morphine decreases MC4 receptor (MC4R) mRNA expression in the NAcc, the periaqueductal gray, and neostriatum (Alvaro et al., 1996), brain regions that modulate drug reward, opiate tolerance, and psychomotor stimulation, respectively (Kalivas and Stewart, 1991; Koob and Bloom, 1988; Wise and Bozarth, 1987). On the other hand, chronic treatment with low doses of morphine or cocaine increases MC4R mRNA in the striatum and hippocampus (Alvaro et al., 2003). Consistent with a role in drug self-administration, central infusion of an MCR agonist decreases the acquisition of heroin self-administration in rats (van Ree et al., 1981). Ethanol has direct effects on central POMC and α-MSH activity (Angelogianni and Gianoulakis, 1993; Rainero et al., 1990). It is therefore possible that MC neuropeptides modulate neurobiological responses to ethanol and participate in the control of voluntary ethanol consumption. Several observations are consistent with this hypothesis. First, α-MSH is expressed in brain regions involved with neurobiological responses to ethanol, including the striatum, NAcc, VTA, amygdala, hippocampus, and hypothalamus (Bloch et al., 1979; Dube et al., 1978; Jacobowitz and O’Donohue, 1978; O’Donohue and Jacobowitz, 1980; O’Donohue et al., 1979; Yamazoe et al., 1984). Second, rats selectively bred for high ethanol drinking (AA [Alko, alcohol]) have low levels of MC3R in the shell of the NAcc but have high levels of MC3R and MC4R in various regions of the hypothalamus, when compared with low–ethanol-drinking rats (Lindblom et al., 2002b). Third, central infusion of MTII significantly reduces voluntary ethanol drinking in AA rats with an established history of ethanol intake (Ploj et al., 2002). Recently, MTII-induced reduction of ethanol consumption was shown to be receptor mediated and not associated with alterations of ethanol metabolism in C57BL/6 mice (Navarro et al., 2003). One objective of the present report was to study the role of selected MCRs in the modulation of MCR agonist–induced reduction of ethanol intake. In rodents, MC peptides act through five receptors (MC1R–MC5R) (Hadley and Haskell-Luevano, 1999). MC3R and MC4R are expressed at high levels in the brain (Alvaro et al., 1997), whereas MC1R and MC5R are detected at low levels and in only limited brain regions (Adan and Gispen, 1997; Barrett et al., 1994; Xia et al., 1995). MTII binds, with varying affinity, to all centrally expressed MCRs (Haskell-Luevano et al., 1997; Schioth et al., 1997). Thus, it is unclear which MCRs are important for modulating MTII-induced reductions of ethanol consumption (Navarro et al., 2003; Ploj et al., 2002). To assess the contribution of MC3R, we examined MTII-induced alteration of ethanol drinking in Mc3r–knock-out (Mc3r−/−) and wild-type (Mc3r+/+) mice. To assess the role of MC4R, we studied ethanol intake by C57BL/6J mice after central infusion of the highly selective MC4R agonist cyclo(NH-CH2-CH2-CO-His-d-Phe-Arg-Trp-Glu)-NH2. A second objective was to determine whether central administration of AgRP-(83-132) would increase ethanol consumption by mice. In vivo, AgRP-(83-132) is a potent and nonselective MCR antagonist (Quillan et al., 1998). Such results would strengthen the argument that MCR signaling plays an important role in the modulation of ethanol self-administration. Because MCR agonists reduce and MCR antagonists increase feeding (Hagan et al., 2000; Hohmann et al., 2000; Hollopeter et al., 1998; Marsh et al., 1999; Thiele et al., 1998), food intake was concurrently assessed in most studies described in the present report.