title: Clodronate inhibits the secretion of proinflammatory cytokines and NO by isolated microglial cells and reduces the number of proliferating glial cells in excitotoxically injured organotypic hippocampal slice cultures.
abstract: Treatment of excitotoxically injured organotypic hippocampal slice cultures (OHSC) with clodronate is known to result in the inhibition of microglial activation. We hypothesized that this is due to direct effects of clodronate on microglial cells, and investigated microglial proliferation in OHSC, and cytokine and NO secretion in isolated microglial cells. N-methyl-D-aspartate (NMDA) lesioning of OHSC resulted in a massive increase in the number of proliferating, bromo-desoxy-uridine (BrdU)-labeled cells that was reduced to control levels after treatment with clodronate (0.1, 1, 10 microg/ml). Triple-labeling revealed that clodronate abrogated the proliferation of both glial fibrillary acidic protein (GFAP)-labeled astrocytes and Griffonia simplicifolia isolectin B4 (IB4)-labeled microglial cells. Furthermore, isolated microglial cells were treated with clodronate after stimulation with lipopolysaccharide (LPS) or macrophage colony stimulating factor (M-CSF). Clodronate (0.01, 0.1, 1 microg/ml) significantly down-regulated the LPS-stimulated microglial secretion of tumor necrosis factor (TNF)-alpha, Interleukin (IL)-1beta and NO, but not of IL-6. In contrast, clodronate significantly reduced the microglial IL-6-release induced by M-CSF, indicating different intracellular pathways. The number and morphology of isolated microglial cells did not change significantly after treatment with clodronate. In summary, the number of proliferating microglial cells and astrocytes after excitotoxic injury is reduced to control levels after treatment with clodronate. Furthermore, clodronate inhibits microglial secretion of proinflammatory cytokines and NO. Clodronate could therefore prove to be a useful tool in the investigation of interactions between damaged neurons and microglial cells.
authors.affiliation: Institute of Anatomy II, Johann Wolfgang Goethe-University, Theodor-Stern-Kai 7, D-60590 Frankfurt am Main, Germany.
authors.forename: Faramarz
authors.initials: F
authors.lastname: Dehghani
authors.forename: Ariane
authors.initials: A
authors.lastname: Conrad
authors.forename: Angelika
authors.initials: A
authors.lastname: Kohl
authors.forename: Horst-Werner
authors.initials: HW
authors.lastname: Korf
authors.forename: Nils P
authors.initials: NP
authors.lastname: Hailer
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D000963:Antimetabolites / Q000494:pharmacology; D001253:Astrocytes / Q000187:drug effects / Q000378:metabolism; D002452:Cell Count; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D004002:Clodronic Acid / Q000494:pharmacology*; D016207:Cytokines / Q000037:antagonists & inhibitors* / Q000378:metabolism; D015536:Down-Regulation / Q000187:drug effects / Q000502:physiology; D005911:Gliosis / Q000139:chemically induced / Q000188:drug therapy* / Q000473:pathology; D006624:Hippocampus / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D066298:In Vitro Techniques; D018836:Inflammation Mediators / Q000494:pharmacology; D007375:Interleukin-1 / Q000037:antagonists & inhibitors / Q000378:metabolism; D017628:Microglia / Q000187:drug effects* / Q000378:metabolism; D016202:N-Methylaspartate; D009498:Neurotoxins; D009569:Nitric Oxide / Q000037:antagonists & inhibitors* / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors / Q000378:metabolism
chemical_list: D000963:Antimetabolites; D016207:Cytokines; D018836:Inflammation Mediators; D007375:Interleukin-1; D009498:Neurotoxins; D014409:Tumor Necrosis Factor-alpha; D004002:Clodronic Acid; D009569:Nitric Oxide; D016202:N-Methylaspartate