N-methyl-D-aspartate receptors mediate endogenous opioid release in enteric neurons after abdominal surgery

Background & Aims: We tested the hypothesis that N-methyl-D-aspartate (NMDA) receptors mediate surgery-induced opioid release in enteric neurons. Methods: We used mu opioid receptor (mu OR) internalization as a measure of opioid release with immunohistochemistry and confocal microscopy. mu OR internalization was quantified in enteric neurons from nondenervated and denervated ileal segments of guinea pig after abdominal laparotomy with and without pretreatment with NMDA-receptor antagonists acting at different recognition sites (+)-5-methyl-10, 11-dihydro-5H-dibenzo [a, b] cyclohepten-5, 10-imine (MK-801) or (D) 2-amino-5-phosphopenoic acid (AP-5) at .5, 1 mg/kg; 8-chloro-4-hydroxy-1-oxo-1, 2-dihydropyridazinol [4,5-]quinoline-5-oxide choline (MRZ 2/576) or 8-chloro-1, 4-dioxo-1,2,3,4-tetrahydropyridazinol [4,5-]quinoline choline salt (MRZ 2/596) at.3, 1 mg/kg, or with an antagonist for the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors, 6-cyano-7-nitroquinoxaline-2, 3-dione (1, 3 mg/kg). To determine whether NMDA stimulation induces opioid release, (1) ilea were exposed to NMDA (100 mu mol/L) and D-serine (10 mu mol/L) with or without the antagonist MK-801 or AP-5 (50 mu mol/L); and (2) neuromuscular preparations of the ileum were stimulated electrically (20 Hz, 20 min) with or without MK-801 or AP-5 (50 mu mol/L). Results: mu OR endocytosis induced by abdominal laparotomy was inhibited significantly by NMDA-receptor antagonists in nondenervated and denervated ileal segments, but not by the AMPA-receptor antagonist. mu OR endocytosis in neurons exposed to NMDA or electrical stimulation was prevented by NMDA-R antagonists. Conclusions: Abdominal laparotomy evokes local release of glutamate that results in endogenous opioid release through the activation of peripheral NMDA receptors. This suggests an interaction between the glutamatergic and opiold systems in response to the noxious and perhaps mechanosensory stimulation of surgery.