Vagal afferent‐dependent cholecystokinin modulation of visceral pain requires central amygdala NMDA‐NR2B receptors in rats

Background Cholecystokinin (CCK), a gut hormone that is released during feeding, exerts gastrointestinal effects in part through vagal pathway. It is reported to be a potential trigger for increased postprandial visceral sensitivity in healthy subjects and, especially in patients with irritable bowel syndrome. NR2B‐containing N‐methyl‐d‐aspartate (NMDA) receptors in the central amygdala (CeA) participate in pain modulation. Systemically administered CCK activates the CeA‐innervating neurons. Here, we investigated whether CCK modulation of visceral sensitivity is mediated through CeA NMDA‐NR2B receptors and whether this modulation involves vagal pathway. Methods We first examined the visceromotor response (VMR) to colorectal distention (CRD) following i.p. injection of CCK octapeptide (CCK‐8) in a rat model. Next, the NR2B antagonist ifenprodil and the NR2A antagonist NVP‐AAM077 were microinjected into the CeA before systemic CCK injection. NR2B phosphorylation was detected by Western blot. To down‐regulate NR2B gene expression, NR2B‐specific small interfering RNA (siRNA) was delivered into CeA neurons by electroporation. In addition, the effects of functional deafferentation by perivagal application of capsaicin and pretreatment with the CCK1 receptor antagonist devazepide were investigated. Key Results CCK‐8 increased VMR to CRD in a dose‐dependent manner. This effect was blunted by intra‐CeA administration of ifenprodil (but not NVP‐AAM077) and was accompanied by phosphorylation of NR2B subunits in the CeA. CCK failed to increase VMR to CRD in NR2B siRNA‐treated rats. Perivagal capsaicin application and pretreatment with devazepide prevented CCK‐induced pronociception and CeA NR2B phosphorylation. Conclusions & Inferences The pronociception induced by systemic CCK, which is vagal afferent‐dependent, requires activation of CeA NMDA‐NR2B receptors. Systemic CCK augments visceral pain response to CRD stimulation in rats. This effect is dependent on vagal afferent activity and the downstream CeA NMDA‐NR2B receptor activation.