Morphine dependence in single enteric neurons from the mouse colon requires deletion of β‐arrestin2

Chronic administration of morphine results in the development of tolerance to the analgesic effects and to inhibition of upper gastrointestinal motility but not to colonic motility, resulting in persistent constipation. In this study we examined the effect of chronic morphine in myenteric neurons from the adult mouse colon. Similar to the ileum, distinct neuronal populations exhibiting afterhyperpolarization (AHP)-positive and AHP-negative neurons were identified in the colon. Acute morphine (3 μM) decreased the number of action potentials, and increased the threshold for action potential generation indicative of reduced excitability in AHP-positive neurons. In neurons from the ileum of mice that were rendered antinociceptive tolerant by morphine-pellet implantation for 5 days, the opioid antagonist naloxone precipitated withdrawal as evidenced by increased neuronal excitability. Overnight incubation of ileum neurons with morphine also resulted in enhanced excitability to naloxone. Colonic neurons exposed to long-term morphine, remained unresponsive to naloxone suggesting that precipitated withdrawal does not occur in colonic neurons. However, morphine-treated colonic neurons from β-arrestin2 knockout mice demonstrated increased excitability upon treatment with naloxone as assessed by change in rheobase, number of action potentials and input resistance. These data suggest that similar to the ileum, acute exposure to morphine in colonic neurons results in reduced excitability due to inhibition of sodium currents. However, unlike the ileum, dependence to chronic exposure of morphine develops in colonic neurons from the β-arrestin2 knockout mice. These studies corroborate the in-vivo findings of the differential role of neuronal β-arrestin2 in the development of morphine tolerance/dependence in the ileum and colon.