Nitric oxide/cyclic GMP signaling regulates motility of a microglial cell line and primary microglia in vitro.
Microglia are the resident immune cells of the brain, which become rapidly activated and migrate to the site of insult in brain infection and disease. Activated microglia generate large amounts of the highly reactive messenger molecule nitric oxide (NO). NO is able to raise cyclic GMP levels via binding to soluble guanylyl cyclase. We investigated potential mechanistic links between inflammation, NO signaling, and microglial migration. To monitor cell migration, we used a scratch wound assay and compared results obtained in the BV-2 microglial line to primary microglia. Incubation with lipopolysaccharide (LPS) as stimulator of acute inflammatory processes enhanced migration of both microglial cell types. LPS activated NO production in BV-2 cells and application of an NO donor increased BV-2 cell migration while an NO scavenger reduced motility. Pharmacological inhibition of soluble guanylyl cyclase and the resulting decrease in motility can be rescued by a membrane permeant analog of cGMP. Despite differences in the threshold towards stimulation with the chemical agents, both BV-2 cells and primary microglia react in a similar way. The important role of NO/cGMP as positive regulator of microglial migration, the downstream targets of the signaling cascade, and resulting cytoskeletal changes can be conveniently investigated in a microglial cell line.
Division of Cell Biology, University of Veterinary Medicine Hannover, Bischofsholer Damm 15/102, D-30173 Hannover, Germany. Electronic address: hannah.scheiblich@tiho-hannover.de.
Hannah
H
Scheiblich
Division of Cell Biology, University of Veterinary Medicine Hannover, Bischofsholer Damm 15/102, D-30173 Hannover, Germany. Electronic address: frank.roloff@tiho-hannover.de.
Frank
F
Roloff
Department of Neurology, Hannover Medical School, Germany; Center for Systems Neuroscience, Hannover, Germany. Electronic address: vikramjeet.singh@med.uni-muenchen.de.
Vikramjeet
V
Singh
Department of Neurology, Hannover Medical School, Germany; Center for Systems Neuroscience, Hannover, Germany. Electronic address: stangel.martin@mh-hannover.de.
Martin
M
Stangel
Division of Cell Biology, University of Veterinary Medicine Hannover, Bischofsholer Damm 15/102, D-30173 Hannover, Germany. Electronic address: michael.stern@tiho-hannover.de.
Michael
M
Stern
Division of Cell Biology, University of Veterinary Medicine Hannover, Bischofsholer Damm 15/102, D-30173 Hannover, Germany; Center for Systems Neuroscience, Hannover, Germany. Electronic address: gerd.bicker@tiho-hannover.de.
Gerd
G
Bicker
D000818:Animals; D002460:Cell Line; D002465:Cell Movement; D006152:Cyclic GMP / Q000378:metabolism*; D007249:Inflammation / Q000378:metabolism; D017628:Microglia / Q000378:metabolism*; D009569:Nitric Oxide / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction
D009569:Nitric Oxide; D006152:Cyclic GMP
BV-2 cells; Cytoskeleton; Inflammation; Primary microglia; iNOS