to two injections versus one, and systemic injections are also accompanied by the stress of unexpected alcohol intoxication, which could in itself induce c-Fos in many areas. To extrapolate, it would appear that endogenous ghrelin may have anxiolytic effects which can be blocked by the antagonist resulting in higher c-Fos levels in the Arc. Lutter et al. (2008) has recently reported that this may be the case after injecting ghrelin and showing less anxiety in the elevated plus maze in mice. While further experimentation is needed to fully explain this phenomenon, this increase in c-Fos immunoreactivity following D-Lys3-GHRP-6 was not specific to the actions of alcohol as it was observed in both ethanol and saline-injected mice. Therefore, it cannot explain the effects of the antagonist on alcohol intake. No potential anxiety effects were observed in pIIIu, as there was a clear interaction between effects of D-Lys3-GHRP-6 and the effects of ethanol on c-Fos induction. This is in agreement with our previous studies showing that c-Fos expression in the pIIIu of C57BL/6J mice is not sensitive to stress or anxiety (Turek and Ryabinin, 2005; Spangler et al., 2009). We conclude therefore, that only the pIIIu, but not the VTA or Arc
