title: Swelling activated Cl- channels in microglia: Biophysics, pharmacology and role in glutamate release.
abstract: Microglia have a swelling-activated Cl- current (which we call IClswell), and while some of its biophysical properties and functional roles have been elucidated, its molecular identity is unknown. To relate this current to cell functions and determine whether it is regulated by mechanisms other than cell swelling, it is important to establish both biophysical and pharmacological fingerprints. Here, we used rat microglia and a cell line derived from them (MLS-9) to study biophysical, regulatory and pharmacological properties of IClswell. The whole-cell current was activated in response to a hypo-osmotic bath solution, but not by voltage, and was time-independent during long voltage steps. The halide selectivity sequence was I->Br->Cl- (Eisenman sequence I) and importantly, the excitatory amino acid, glutamate was permeant. Current activation required internal ATP, and was not affected by the guanine nucleotides, GTPS or GDPS, or physiological levels of internal Mg2+. The same current was activated by a low intracellular ionic strength solution without an osmotic gradient. IClswell was reversibly inhibited by known Cl- channel blockers (NPPB, flufenamic acid, glibenclamide, DCPIB), and by the glutamate release inhibitor, riluzole. Cell swelling evoked glutamate release from primary microglia and MLS-9 cells, and this was inhibited by the blockers (above), and by IAA-94, but not by tamoxifen or the Na+/K+/Cl- symport inhibitor, bumetanide. Together, these results confirm the similarity of IClswell in the two cell types, and point to a role for this channel in inflammation-mediated glutamate release in the CNS.
authors.affiliation: Toronto Western Research Institute, University Health Network, Department of Physiology, University of Toronto, Toronto, Ontario, Canada. schlicht@uhnres.utoronto.ca
authors.forename: Lyanne C
authors.initials: LC
authors.lastname: Schlichter
authors.forename: Timothy
authors.initials: T
authors.lastname: Mertens
authors.forename: Baosong
authors.initials: B
authors.lastname: Liu
mesh_terms: D000255:Adenosine Triphosphate / Q000737:chemistry; D000818:Animals; D001703:Biophysics / Q000379:methods; D001921:Brain / Q000378:metabolism; D002712:Chlorides / Q000737:chemistry; D018698:Glutamic Acid / Q000378:metabolism*; D007477:Ions / Q000737:chemistry; D017628:Microglia / Q000378:metabolism*; D008954:Models, Biological; D009474:Neurons / Q000378:metabolism; D009711:Nucleotides / Q000737:chemistry; D009995:Osmosis; D018408:Patch-Clamp Techniques; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D002712:Chlorides; D007477:Ions; D009711:Nucleotides; D018698:Glutamic Acid; D000255:Adenosine Triphosphate