Whole cell patch-clamp recordings were made from cultured myenteric neurons taken from murine proximal colon. The micropipette contained Cs⁺ to remove K⁺ currents. Depolarization elicited a slowly activating time-dependent outward current (I_tdo), whereas repolarization was followed by a slowly deactivating tail current (I_tail). I_tdo and I_tail were present in ~70% of neurons. We identified these currents as Cl currents (I_Cl), because changing the transmembrane Cl gradient altered the measured reversal potential (E_rev) of both I_tdo and I_tail with that for I_tail shifted close to the calculated Cl equilibrium potential (E_Cl). I_Cl are Ca²⁺-activated Cl current [I_Cl(Ca)] because they were Ca²⁺ dependent. E_Cl, which was measured from the E_rev of I_Cl(Ca) using a gramicidin perforated patch, was 33 mV. This value is more positive than the resting membrane potential (56.3 ± 2.7 mV), suggesting myenteric neurons accumulate intracellular Cl. -Conotoxin GIVA [0.3 µM; N-type Ca²⁺ channel blocker] and niflumic acid [10 µM; known I_Cl(Ca) blocker], decreased the I_Cl(Ca). In conclusion, these neurons have I_Cl(Ca) that are activated by Ca²⁺ entry through N-type Ca²⁺ channels. These currents likely regulate postspike frequency adaptation.