Signaling mechanisms of interferon gamma induced apoptosis in chromaffin cells: involvement of nNOS, iNOS, and NFκB

• Published in: Journal of neurochemistry 2009-02, Vol.108 (4), p.1083-1096
• Main Authors: Pérez-Rodríguez, Rocío, Roncero, Cesáreo, Oliván, Ana María, González, María Pilar, Oset-Gasque, María Jesús
• Format: Article
• Language: English
• Subjects: apoptosis; cell death; chromaffin cells; JAK/STAT; nitric oxide synthase; nuclear factor κB; signal transduction
• ISSN: 0022-3042; 1471-4159
• DOI: 10.1111/j.1471-4159.2008.05862.x

Previous work of our group stated that exogenously added and endogenous nitric oxide (NO) generated by cytokines induce apoptosis in chromaffin cells. In this work, we investigate the specific regulation of the NO synthase (NOS) isoforms, inducible NOS (iNOS) and neuronal NOS (nNOS), and their particular participation in cell death induced by interferon gamma (IFNγ). Lipopolysaccharide (LPS) and IFNγ increase iNOS expression, with no effect on nNOS expression. On the other hand, dexamethasone increases basal nNOS expression but decreases LPS + IFNγ-induced iNOS expression. IFNγ-induced cell death was abolished by W-1400, a specific iNOS inhibitor, but only partially by nNOS inhibitors [N-ω-propyl- l-arginine (N-PLA), 3-Bromo-7-nitroindazol (7-NI), l-methyl thiocitrulline and N-methyl l-arginine], indicating the main iNOS participation in chromaffin cell death. IFNγ and LPS induce nuclear factor κB (NFκB) translocation to the nucleus, a process implicated in activation of iNOS expression, as inhibition of NFκB translocation, by SN50, decreased iNOS expression. In addition, IFNγ and LPS induce ⁸⁴⁷Ser-nNOS phosphorylation, inhibiting nNOS activity. Both processes, nNOS phosphorylation and iNOS expression induced by LPS + IFNγ, are regulated by Janus Kinase/Signal Transducer and Activator of Transcription (JAK/STAT) pathway, as IFNγ increases ⁷²⁷STAT-3 phosphorylation and specific inhibitors of JAK/STAT pathway, such as AG490, inhibited both processes. Taken together, these results support the hypothesis of an inactivating phosphorylation of nNOS by IFNγ, via JAK/STAT, in bovine chromaffin cells. Low NO concentrations achieved by this event, would activate NFκB translocation, increasing iNOS expression and generating, this last, high apoptotic NO concentrations.