title: Interleukin-1beta exacerbates and interleukin-1 receptor antagonist attenuates neuronal injury and microglial activation after excitotoxic damage in organotypic hippocampal slice cultures.
abstract: The effects of interleukin (IL)-1beta and IL-1 receptor antagonist (IL-1ra) on neurons and microglial cells were investigated in organotypic hippocampal slice cultures (OHSCs). OHSCs obtained from rats were excitotoxically lesioned after 6 days in vitro by application of N-methyl-D-aspartate (NMDA) and treated with IL-1beta (6 ng/mL) or IL-1ra (40, 100 or 500 ng/mL) for up to 10 days. OHSCs were then analysed by bright field microscopy after hematoxylin staining and confocal laser scanning microscopy after labeling of damaged neurons with propidium iodide (PI) and fluorescent staining of microglial cells. The specificity of PI labeling of damaged neurons was validated by triple staining with neuronal and glial markers and it was observed that PI accumulated in damaged neurons only but not in microglial cells or astrocytes. Treatment of unlesioned OHSCs with IL-1beta did not induce neuronal damage but caused an increase in the number of microglial cells. NMDA lesioning alone resulted in a massive increase in the number of microglial cells and degenerating neurons. Treatment of NMDA-lesioned OHSCs with IL-1beta exacerbated neuronal cell death and further enhanced microglial cell numbers. Treatment of NMDA-lesioned cultures with IL-1ra significantly attenuated NMDA-induced neuronal damage and reduced the number of microglial cells, whereas application of IL-1ra in unlesioned OHSCs did not induce significant changes in either cell population. Our findings indicate that: (i) IL-1beta directly affects the central nervous system and acts independently of infiltrating hematogenous cells; (ii) IL-1beta induces microglial activation but is not neurotoxic per se; (iii) IL-1beta enhances excitotoxic neuronal damage and microglial activation and (iv) IL-1ra, even when applied for only 4 h, reduces neuronal cell death and the number of microglial cells after excitotoxic damage.
authors.affiliation: Dr Senckenbergische Anatomie, Institute of Anatomy 2, Johann Wolfgang Goethe-University, D-60590 Frankfurt am Main, Federal Republic of Germany.
authors.forename: Nils P
authors.initials: NP
authors.lastname: Hailer
authors.forename: Cornelia
authors.initials: C
authors.lastname: Vogt
authors.forename: Horst-Werner
authors.initials: HW
authors.lastname: Korf
authors.forename: Faramarz
authors.initials: F
authors.lastname: Dehghani
mesh_terms: D000818:Animals; D001930:Brain Injuries / Q000188:drug therapy / Q000473:pathology; D002450:Cell Communication / Q000187:drug effects; D016923:Cell Death / Q000187:drug effects; D004396:Coloring Agents; D006624:Hippocampus / Q000187:drug effects / Q000473:pathology*; D053590:Interleukin 1 Receptor Antagonist Protein; D007375:Interleukin-1 / Q000494:pharmacology*; D017628:Microglia / Q000187:drug effects* / Q000502:physiology; D018613:Microscopy, Confocal; D009474:Neurons / Q000166:cytology* / Q000187:drug effects; D009498:Neurotoxins / Q000633:toxicity; D009924:Organ Culture Techniques; D037121:Plant Lectins; D011419:Propidium; D051381:Rats; D017208:Rats, Wistar; D012795:Sialoglycoproteins / Q000494:pharmacology*
chemical_list: D004396:Coloring Agents; C045475:Griffonia simplicifolia lectins; D053590:Interleukin 1 Receptor Antagonist Protein; D007375:Interleukin-1; D009498:Neurotoxins; D037121:Plant Lectins; D012795:Sialoglycoproteins; D011419:Propidium