title: Microglial SK3 and SK4 currents and activation state are modulated by the neuroprotective drug, riluzole.
abstract: Microglia monitor the CNS for 'danger' signals after acute injury, such as stroke and trauma, and then undergo complex activation processes. Classical activation of microglia can produce neurotoxic levels of glutamate and immune mediators (e.g., pro-inflammatory cytokines, reactive oxygen and nitrogen species), while alternative activation up-regulates anti-inflammatory molecules and is thought to resolve inflammation and protect the brain. Thus, pharmacological strategies to decrease classical- and/or promote alternative activation are of interest. Here, we assessed actions of the neuroprotective drug, riluzole, on two Ca(2+)- activated K channels in microglia - SK3 (KCa2.3, KCNN3) and SK4 (KCa3.1, KCNN4) - and on classical versus alternative microglial activation. Riluzole is used to treat amyotrophic lateral sclerosis, and is in clinical trials for several other CNS disorders, where it has been presumed to target neurons and reduce glutamate-mediated toxicity. We show that simply elevating intracellular Ca(2+) to micromolar levels in whole-cell recordings does not activate SK channels in a cell line derived from primary rat microglia (MLS-9). In intact cells, riluzole raised cytoplasmic Ca(2+), but it was marginal (~200 nM) and transient (2 min). Surprisingly then, in whole cell recordings, riluzole rapidly activated SK3 and SK4 channels for as long as it was present, and did not require elevated intracellular Ca(2+). We then used primary rat microglia to analyze expression of several activation markers and inflammatory mediators. Riluzole decreased classical LPS-induced activation, and increased some aspects of IL-4-induced alternative activation. These actions on microglia suggest an additional mechanism underlying the neuroprotective actions of riluzole.
authors.affiliation: Toronto Western Research Institute, University Health Network, MC9-417, 399 Bathurst Street, Toronto, Ontario M5T 2S8, Canada.
authors.forename: B-S
authors.initials: BS
authors.lastname: Liu
authors.forename: R
authors.initials: R
authors.lastname: Ferreira
authors.forename: S
authors.initials: S
authors.lastname: Lively
authors.forename: L C
authors.initials: LC
authors.lastname: Schlichter
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D020013:Calcium Signaling / Q000187:drug effects / Q000502:physiology; D002460:Cell Line; D015847:Interleukin-4 / Q000494:pharmacology; D051660:Intermediate-Conductance Calcium-Activated Potassium Channels / Q000187:drug effects*; D008070:Lipopolysaccharides / Q000494:pharmacology; D017628:Microglia / Q000187:drug effects* / Q000378:metabolism; D018696:Neuroprotective Agents / Q000494:pharmacology*; D018408:Patch-Clamp Techniques; D051381:Rats; D060888:Real-Time Polymerase Chain Reaction; D019782:Riluzole / Q000494:pharmacology*; D051640:Small-Conductance Calcium-Activated Potassium Channels / Q000187:drug effects*
chemical_list: D051660:Intermediate-Conductance Calcium-Activated Potassium Channels; C496901:Kcnn3 protein, rat; C491575:Kcnn4 protein, rat; D008070:Lipopolysaccharides; D018696:Neuroprotective Agents; D051640:Small-Conductance Calcium-Activated Potassium Channels; D015847:Interleukin-4; D019782:Riluzole; D002118:Calcium