In vitro reactive nitrating species toxicity in dissociated spinal motor neurons from NFL (-/-) and hNFL (+/+) transgenic mice.
We utilized fetal spinal motor neurons isolated from either NFL (-/-) or hNFL (+/+) transgenic mice to determine whether the loss of the low molecular weight neurofilament protein (NFL) places spinal motor neurons at a greater risk for cell death triggered by reactive nitrating species (RNS). After 21 days in serum-free, antibiotic-free medium, both the NFL (-/-) and hNFL (+/+) motor neurons developed neurofilamentous aggregates. Cultures were then exposed to nitric oxide(100 microM NOC 5, 100 microM NOC 12, or 2 mM sodium nitroprusside) or to peroxynitrite (250 mM SIN-1) forvarying intervals. NFL (-/-) cultures demonstrated extensive numbers of apoptotic neurons within six hours and complete cell loss by 24 hours in response to NOC 5 and NOC 12. In contrast, apoptosis was only observed in the motor neurons derived from control (C57bl/6) or hNLF (+/+) mice at 24 hours. In response to 2 mM sodium nitroprusside, necrosis was induced in all cells within 60 minutes. In response to 250 mM SIN-1, both C57bl/6 and hNFL (+/+) cells survived to six hours with only minimal evidence of degeneration while NFL (-/-) motor neurons were necrotic by 60 minutes. These observations suggest that NFL deficient motor neurons are at an enhanced risk of cell death mediated by RNS.
D000818:Animals; D016923:Cell Death / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D006801:Humans; D066298:In Vitro Techniques; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008981:Molsidomine / Q000031:analogs & derivatives* / Q000494:pharmacology; D009046:Motor Neurons / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D016900:Neurofilament Proteins / Q000235:genetics*; D009569:Nitric Oxide / Q000378:metabolism; D020030:Nitric Oxide Donors / Q000494:pharmacology; D009599:Nitroprusside / Q000494:pharmacology; D009603:Nitroso Compounds / Q000494:pharmacology; D030421:Peroxynitrous Acid / Q000378:metabolism; D013116:Spinal Cord / Q000166:cytology; D013482:Superoxide Dismutase / Q000378:metabolism; D014226:Triazenes / Q000494:pharmacology
C114535:1-hydroxy-2-oxo-3-(3-aminopropyl)-3-isopropyl-1-triazene; C118603:NOC 12; D016900:Neurofilament Proteins; D020030:Nitric Oxide Donors; D009603:Nitroso Compounds; D014226:Triazenes; C044882:neurofilament protein L; D030421:Peroxynitrous Acid; D009599:Nitroprusside; D009569:Nitric Oxide; C002385:linsidomine; D008981:Molsidomine; D013482:Superoxide Dismutase