| Abstrakt: |
Imredy JP, Penniman JR, Dech SJ, Irving WD, Salata JJ. Modeling of the adrenergic response of the human I[subκs] current (hKCNQIIhKCNEI) stably expressed in HEK-293 cells. Am J Physiol Heart Circ Physiol 295: H 1867-HI 881, 2008. First published August 29, 2008; doi: 10.1152/ajpheart.433.2008.-Stable coexpression of human (h)KCNQ1 and hKCNE1 in human embryonic kidney (HEK)-293 cells reconstitutes a nativelike slowly activating delayed rectifier K[sup+] current (HEK-I[subκs]), allowing β-adrenergic modulation of the current by stimulation of endogenous receptors in the host cell line. HEK-I[subκs]. was enhanced twoto fourfold by isoproterenol (EC50 = 13 nM), forskolin (10 μM), or 8-(4-chlorophenylthio)adenosine 3',S'-cyclic monophosphate (50 μM), indicating an intact cAMP-dependent ion channel-regulating pathway analogous to the PKA-dependent regulation observed in native cardiac myocytes. Activation kinetics of HEK-I[subκs]. were accurately fit with a novel modified second-order Hodgkin-Huxley (H-H) gating model incorporating a fast and a slow gate, each independent of each other in scale and adrenergic response, or a "heterodimer" model. Macroscopically, β-adrenergic enhancement shifted the current activation threshold to more negative potentials and accelerated activation kinetics while leaving deactivation kinetics relatively unaffected. Modeling of the current response using the H-H model indicated that observed changes in gating could be explained by modulation of the opening rate of the fast gate. Under control conditions at nearly physiological temperatures (35°C), rate-dependent accumulation of HEK-I[subκs] was observed only at pulse frequencies exceeding 3 Hz. Rate-dependent accumulation of I[subκs] at high pulsing rate had two phases, an initial staircaselike effect followed by a slower, incremental accumulation phase. These phases are readily interpreted in the context of a heterodimenc H-H model with two independent gates with differing closing rates. In the presence of isoproterenol after normalizing for its tonic effects, rate-dependent accumulation of HEK-l[subκs] appeared at lower pulse frequencies and was slightly enhanced (-25%) over control. [ABSTRACT FROM AUTHOR] |
