title: Induction of potassium channels in mouse brain microglia: cells acquire responsiveness to pneumococcal cell wall components during late development.
abstract: Lipopolysaccharides derived from cell walls of Gram-negative bacteria have proven a useful tool to simulate bacterial infection of the central nervous system. Rapid activation of microglia within the brain parenchyma as well as in vitro has thereby been shown to be an early event upon bacterial or lipopolysaccharide challenges. Less is known about microglial responses to a contact with Gram-positive bacteria, such as Streptococcus pneumoniae, a lethal pathogen causing meningitis with a 30% mortality rate. In the present study, we compared lipopolysaccharide-induced microglial activation in vitro with that induced by preparations of pneumococcal cell walls. As a readout of microglial activation, we studied by patch-clamp recording the expression of outward rectifying potassium currents (IK+OR), which are known to be induced by lipopolysaccharide. We found that pneumococcal cell walls and lipopolysaccharide induced a similar type of IK+OR. Stimulation of IK+OR by pneumococcal cell walls and lipopolysaccharide involved protein synthesis since it was not induced in the presence of cycloheximide. Pharmacological characterization of the pneumococcal cell wall- and lipopolysaccharide-induced currents with specific ion channel blockers indicated for both cases expression of the charybdotoxin/margatoxin-sensitive Kv1.3 subtype of the Shaker family of voltage-dependent potassium channels. Activation of the outward currents by pneumococcal cell walls depended on the developmental stage: while lipopolysaccharide triggered IK+OR in both embryonal and postnatal microglial cells, pneumococcal cell walls had only a marginal effect on embryonal cells. This, however, does not imply that embryonic microglial cells are unresponsive to pneumococcal cell walls. In both embryonic and postnatal cells, (i) the amplitude of the constitutively expressed inward rectifying potassium current was significantly reduced, (ii) tumor necrosis factor-a was released and (iii) the cells changed their morphology, similarly as it was induced by lipopolysaccharide treatment. Thus, embryonic microglial cells are sensitive to pneumococcal cell wall challenges, but respond with a distinctly different pattern of physiological reactions. The expression of IK+OR could thus be a suitable tool to study signalling cascades selectively involved in the activation of microglia by Gram-negative and -positive cell wall components and to functionally distinguish between populations of microglial cells.
authors.affiliation: Department of CNS Research, Boehringer Ingelheim Pharma KG, Ingelheim, Germany.
authors.forename: H J
authors.initials: HJ
authors.lastname: Draheim
authors.forename: M
authors.initials: M
authors.lastname: Prinz
authors.forename: J R
authors.initials: JR
authors.lastname: Weber
authors.forename: T
authors.initials: T
authors.lastname: Weiser
authors.forename: H
authors.initials: H
authors.lastname: Kettenmann
authors.forename: U K
authors.initials: UK
authors.lastname: Hanisch
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D001030:Apamin / Q000494:pharmacology; D001921:Brain / Q000166:cytology / Q000276:immunology / Q000502:physiology*; D002473:Cell Wall / Q000276:immunology*; D002478:Cells, Cultured; D018999:Charybdotoxin / Q000494:pharmacology; D004546:Elapid Venoms / Q000494:pharmacology; D004622:Embryo, Mammalian; D051664:Kv1.3 Potassium Channel; D008070:Lipopolysaccharides / Q000494:pharmacology; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D051379:Mice; D008815:Mice, Inbred Strains; D017628:Microglia / Q000166:cytology / Q000276:immunology / Q000502:physiology*; D009498:Neurotoxins / Q000494:pharmacology; D026902:Potassium Channel Blockers; D015221:Potassium Channels / Q000096:biosynthesis* / Q000502:physiology; D024683:Potassium Channels, Tandem Pore Domain; D024642:Potassium Channels, Voltage-Gated; D051381:Rats; D012604:Scorpion Venoms; D051637:Shaker Superfamily of Potassium Channels; D013296:Streptococcus pneumoniae / Q000276:immunology*
chemical_list: D004546:Elapid Venoms; C496974:Kcna3 protein, mouse; C496975:Kcna3 protein, rat; D051664:Kv1.3 Potassium Channel; D008070:Lipopolysaccharides; D009498:Neurotoxins; D026902:Potassium Channel Blockers; D015221:Potassium Channels; D024683:Potassium Channels, Tandem Pore Domain; D024642:Potassium Channels, Voltage-Gated; D012604:Scorpion Venoms; D051637:Shaker Superfamily of Potassium Channels; D018999:Charybdotoxin; D001030:Apamin; C077910:margatoxin; C020273:dendrotoxin