p75 neurotrophin receptor mediates neurotrophin activation of NF-kappa B and induction of iNOS expression in P19 neurons.
P19 embryonic carcinoma cells can be differentiated into neurons that form synaptic connections and that produce a variety of neurotransmitters. Results of RT-PCR indicate that P19 neurons express several neurotrophin receptors (p75(NTR), trkB, and trkC, but not trkA) but they do not express any of the four neurotrophins. Consistent with the presence of trkB but not trkA, BDNF causes rapid phosphorylation of MAP kinases ERK1 and ERK2, but NGF does not. Neurotrophins induce translocation of NF-kappaB into the nucleus. All four neurotrophins induce activation of NF-kappaB in a biphasic manner. This effect is apparently mediated by p75(NTR), because an inhibitor of trk receptors, K252a, does not inhibit activation of NF-kappaB. Instead, K252a itself promotes activation of NF-kappaB and this effect is additive with the effect of neurotrophins. Inhibition of reactive oxygen intermediates with PDTC completely abolishes basal activity of NF-kappaB and strongly inhibits activation of NF-kappaB by neurotrophins, indicating an important role of reactive oxygen intermediates in the pathway by which neurotrophins activate NF-kappaB. NF-kappaB is known to promote expression of the iNOS gene. We found that all four neurotrophins increased iNOS mRNA levels, resulting in increased accumulation of iNOS protein. In contrast, none of the neurotrophins stimulated nNOS mRNA or protein synthesis. PDTC abolishes constitutive and neurotrophin-induced expression of iNOS mRNA and protein and abolishes constitutive expression of nNOS mRNA, suggesting that reactive oxygen intermediates promote expression of nNOS.
Burke
Melanie A
MA
Department of Neurology, Box 356465, University of Washington School of Medicine, Seattle, Washington 98195, USA. mab@u.washington.edu
Bothwell
Mark
M
D000818:Animals; D004305:Dose-Response Relationship, Drug; D004790:Enzyme Induction / Q000502:physiology; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D051379:Mice; D016328:NF-kappa B / Q000378:metabolism*; D009414:Nerve Growth Factors / Q000378:metabolism / Q000502:physiology*; D009474:Neurons / Q000378:metabolism* / Q000502:physiology; D019001:Nitric Oxide Synthase / Q000096:biosynthesis*; D052247:Nitric Oxide Synthase Type II; D020800:Receptor, Nerve Growth Factor; D017475:Receptors, Nerve Growth Factor / Q000037:antagonists & inhibitors / Q000502:physiology*; D015398:Signal Transduction / Q000502:physiology; D014407:Tumor Cells, Cultured
D016328:NF-kappa B; D009414:Nerve Growth Factors; D020800:Receptor, Nerve Growth Factor; D017475:Receptors, Nerve Growth Factor; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse