Acute pharmacological studies have implicated mu-opioid receptors (MORs) in the shell of the nucleus accumbens (NAC) in mediating responses for palatable food and other natural and drug-induced rewards. However, the long-term behavioral effects of inactivating signal transduction via accumbal MORs, as quantified by an anatomically defined loss of cellular activity, have never been analysed. We combined microinfusion of the irreversible MOR antagonist, β-funaltrexamine (β-FNA; 8.0 nmol/0.8 µL, n = 9; controls, n = 6) with mapping by [35S]GTPγS autoradiography to demonstrate an anatomically specific loss of the coupling of MORs to their G-proteins in the dorsal caudomedial shell of the NAC in rabbits. β-FNA did not alter the stimulated coupling of kappa-opioid receptors. This selective blockade of the cellular function of MORs persistently decreased consumption of a palatable sucrose solution by 40% during a daily 4-h test conducted 2, 3 and 4 days after infusion. β-FNA did not alter body weight or 20-h consumption of standard chow or water. In 10 different rabbits, infusion of the selective, competitive MOR antagonist, CTAP (D-Phe-Cys-Tyr-D-Trp-Arg-Thr-Pen-Thr-NH2) into the same locus produced a reversible decrease in sucrose consumption, with normal intakes returning on the next day. Together, these data appear to establish that MORs in this accumbal subregion support responding for orosensory reward. Overall, these results visualize a discrete brain locus where cellular actions of endogenous opioids mediate behaviors involved in self-administration of foods and perhaps other hedonically valued substances, such as ethanol and drugs of abuse.