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The properties of the hyperpolarization-activated inward cation current (Ih) in mouse anterior pituitary, AtT20 D16:16 corticotropes was characterized by whole cell patch clamp recording. In response to hyperpolarizing steps a large, slowly activating, voltage-dependent inward current was activated with a half maximal activation voltage (V0.5) of -96.2+/-3.1 mV with a time constant of 168+/-13 msec determined at -140 mV at room temperature. Ih had a reversal potential of -35.5+/-1.0 mV and -23.3+/-1.4 mV using 5 mM and 25 mM extracellular K+, respectively, with a relative permeability ratio for Na+ and K+ of 0.24. The current was completely blocked by 2 mM extracellular CsCl and partially blocked by ZD7288 (100 microM) but was unaffected by TEA (10 mM) or Ba2+ (1 mM). RT-PCR analysis revealed robust expression of HCN1, but not HCN2 or HCN3, subunits of hyperpolarization-activated cation channels. The endogenous Ih current was weakly activated by cAMP but robustly inhibited by the cAMP antagonist, Rp-8-CPT-cAMPS. Activation or suppression of protein kinase C activity had no significant effect on the Ih current. The data suggest that in AtT20 D16:16 corticotropes Ih is tonically regulated by the cAMP-signaling cascade and may serve to limit excessive hyperpolarization.

Characterization of Hyperpolarization-Activated Cation Currents in Mouse Anterior Pituitary, AtT20 D16:16 Corticotropes*