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. Author manuscript; available in PMC: 2012 Mar 1.

Published in final edited form as: Exp Physiol. 2010 Dec 17;96(3):305–316. doi: 10.1113/expphysiol.2010.055038

A. Isolated colonic crypts were stimulated by adrenaline [10μM] during standard whole-cell patch-clamp recording (see Methods): adrenaline (solid line), basal level (dashed line). Increased Cl⁻ current was apparent at the K⁺ reversal potential (E_K). B. Adrenaline stimulated current with pipet solution K⁺ replaced with Cs⁺, consistent with activated Cl⁻ current. Addition of CaCCinh-A01 [50μM] reduced current to basal levels. C. Adrenaline stimulated (*) inward current measured at E_K (−86mV) with an activation τ_0.5 of ~14sec (I_Cl, mean±sem, n=17). Variable delay in onset was adjusted by aligning the times at which I_Cl began increasing. Similar activation occurred in the absence of K⁺ (CsCl pipet, n=8). Substitution of Cl⁻ in the recording solutions eliminated the response (n=4); currents were aligned at the time of adrenaline addition. D. Cl⁻ channel blockers (#) inhibited the adrenaline-activated inward current (added 3–5min earlier). The τ_0.5 of inhibition was ~20sec for CaCCinh-A01 (50μM, n=3), ~18sec for GlyH-101 (50μM, n=3), and ~30 sec for NPPB (100μM, n=3).

A. Isolated colonic crypts were stimulated by adrenaline [10μM] during standard whole-cell patch-clamp recording (see Methods): adrenaline (solid line), basal level (dashed line). Increased Cl⁻ current was apparent at the K⁺ reversal potential (E_K). B. Adrenaline stimulated current with pipet solution K⁺ replaced with Cs⁺, consistent with activated Cl⁻ current. Addition of CaCCinh-A01 [50μM] reduced current to basal levels. C. Adrenaline stimulated (*) inward current measured at E_K (−86mV) with an activation τ_0.5 of ~14sec (I_Cl, mean±sem, n=17). Variable delay in onset was adjusted by aligning the times at which I_Cl began increasing. Similar activation occurred in the absence of K⁺ (CsCl pipet, n=8). Substitution of Cl⁻ in the recording solutions eliminated the response (n=4); currents were aligned at the time of adrenaline addition. D. Cl⁻ channel blockers (#) inhibited the adrenaline-activated inward current (added 3–5min earlier). The τ_0.5 of inhibition was ~20sec for CaCCinh-A01 (50μM, n=3), ~18sec for GlyH-101 (50μM, n=3), and ~30 sec for NPPB (100μM, n=3).