Inverse Modulation of Neuronal Kv12.1 and Kv11.1 Channels by 4-Aminopyridine and NS1643

Marlen Dierich; Saskia Evers; Bettina U Wilke; Michael G Leitner

doi:10.3389/fnmol.2018.00011

Inverse Modulation of Neuronal K_v12.1 and K_v11.1 Channels by 4-Aminopyridine and NS1643

Front Mol Neurosci. 2018 Jan 30:11:11. doi: 10.3389/fnmol.2018.00011. eCollection 2018.

Authors

Marlen Dierich¹, Saskia Evers¹, Bettina U Wilke¹, Michael G Leitner^{1

2}

Affiliations

¹ Department of Neurophysiology, Institute of Physiology and Pathophysiology, Philipps University of Marburg, Marburg, Germany.
² Division of Physiology, Department of Physiology and Medical Physics, Innsbruck Medical University, Innsbruck, Austria.

Abstract

The three members of the ether-à-go-go-gene-like (Elk; K_v12.1-K_v12.3) family of voltage-gated K⁺ channels are predominantly expressed in neurons, but only little information is available on their physiological relevance. It was shown that K_v12.2 channels modulate excitability of hippocampal neurons, but no native current could be attributed to K_v12.1 and K_v12.3 subunits yet. This may appear somewhat surprising, given high expression of their mRNA transcripts in several brain areas. Native K_v12 currents may have been overlooked so far due to limited knowledge on their biophysical properties and lack of specific pharmacology. Except for K_v12.2, appropriate genetically modified mouse models have not been described; therefore, identification of K_v12-mediated currents in native cell types must rely on characterization of unique properties of the channels. We focused on recombinant human K_v12.1 to identify distinct properties of these channels. We found that K_v12.1 channels exhibited significant mode shift of activation, i.e., stabilization of the voltage sensor domain in a "relaxed" open state after prolonged channel activation. This mode shift manifested by a slowing of deactivation and, most prominently, a significant shift of voltage dependence to hyperpolarized potentials. In contrast to related K_v11.1, mode shift was not sensitive to extracellular Na⁺, which allowed for discrimination between these isoforms. Sensitivity of K_v12.1 and K_v11.1 to the broad-spectrum K⁺ antagonist 4-aminopyridine was similar. However, 4-AP strongly activated K_v12.1 channels, but it was an inhibitor of K_v11 channels. Interestingly, the agonist of K_v11 channels NS1643 also differentially modulated the activity of these channels, i.e., NS1643 activated K_v11.1, but strongly inhibited K_v12.1 channels. Thus, these closely related channels are distinguished by inverse pharmacological profiles. In summary, we identified unique biophysical and pharmacological properties of K_v12.1 channels and established straightforward experimental protocols to characterize K_v12.1-mediated currents. Identification of currents in native cell types with mode shift that are activated through 4-AP and inhibited by NS1643 can provide strong evidence for contribution of K_v12.1 to whole cell currents.

Keywords: 4-aminopyridine; HERG; Kv10; Kv11; Kv12; NS1643; mode shift; voltage-dependent potentiation.