Macrophage migration inhibitory factor (MIF) increases neuronal delayed rectifier K current

Macrophage migration inhibitory factor (MIF) has widespread actions in the immune, endocrine and nervous systems. Previously, we reported that increases in the intracellular levels of MIF depress the firing of hypothalamus/brainstem neurons in culture, including the chronotropic actions of angiotensin II (Ang II). The objective of this study was to investigate the effects of MIF on delayed rectifier K current (IKv), one of the component currents whose activity contributes to neuronal firing. Intracellular perfusion of MIF (80 nM) into Sprague Dawley rat neuronal cultures caused a significant increase in IKv, as measured by patch-clamp recordings. This effect was apparent by 3 minutes, and was maximal after 20-30 minutes. IKv current density (pA/pF) increased from 31.58 ± 2.36 in controls to 41.88 ± 3.76 in MIF-treated neurons (mean ± SE; n=9; p<0.01). MIF that had been inactivated by boiling did not alter IKv, and MIF-neutralizing antibodies abolished the action of rMIF. The stimulatory effect of MIF on IKv current density was mimicked by intracellular application of either P1S-MIF (80 nM) or the peptide MIF-(5065) [0.8-8 μM], both of which harbor the thiol-protein oxidoreductase (TPOR) activity of the MIF molecule. Conversely, neither C60S-MIF (80 nM) nor the MIF homologue Ddopachrome tautomerase (DCT, 80 nM), both of which lack TPOR activity, altered IKv. Lastly, the increase in IKv produced by rMIF was abolished by the superoxide scavenger Tiron (1 mM). These studies indicate that the neuronal action of MIF includes a stimulatory action on IKv that may be mediated via a TPOR/superoxide-scavenging mechanism. JN-00499-2005.R2