title: Laminin activates NF-kappaB in Schwann cells to enhance neurite outgrowth.
abstract: Extracellular matrix (ECM) molecules and Schwann cells (SCs) are important components of peripheral nerve regeneration. In this study, the role of the transcription factor nuclear factor kappa B (NF-kappaB) in SC activation in response to laminin and the subsequent effect on in vitro neurite outgrowth was investigated. Immunocytochemistry and Western blot analysis showed that compared with poly-d-lysine (PDL), laminin enhanced the phosphorylation of IkappaB and p65 NF-kappaB signalling proteins in SCs. Phospho NF-kappaB-p65 was localised to the nucleus indicating activation of NF-kappaB. To assess the functional effect of NF-kappaB activation, SCs plated on PDL or laminin were pre-treated with NF-kappaB inhibitors, 6-amino-4-(4-phenoxyphenylethylamino)quinazoline (QNZ) or Z-leu-leu-leu-CHO (MG-132) before NG108-15 neuronal cells were seeded on the SC monolayer. After 24h co-culture in the absence of inhibitors, SCs seeded on laminin enhanced the mean number and length of neurites extended by NG108-15 cells (1.87+/-0.13 neurites; 238.74+/-8.53microm) compared with those cultured in the presence of SCs and PDL (1.26+/-0.07 neurites; 157.57+/-9.80microm). At 72h, neurite length had further increased to 321.83+/-6.60microm in the presence of SCs and laminin. Inhibition of NF-kappaB completely abolished the effect of laminin on SC evoked neurite outgrowth at 24h and reduced the enhancement of neurite length by over 60% at 72h. SC proliferation was unaffected by NF-kappaB inhibition suggesting that the NF-kappaB signalling pathway plays a discrete role in the activation of SCs and their neurotrophic potential.
authors.affiliation: Blond McIndoe Laboratories, The University of Manchester, Manchester, UK.
authors.forename: Stephanie J
authors.initials: SJ
authors.lastname: Armstrong
authors.forename: Mikael
authors.initials: M
authors.lastname: Wiberg
authors.forename: Giorgio
authors.initials: G
authors.lastname: Terenghi
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Kingham
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D002478:Cells, Cultured; D004347:Drug Interactions; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D066298:In Vitro Techniques; D007797:Laminin / Q000494:pharmacology*; D007976:Leupeptins / Q000494:pharmacology; D016328:NF-kappa B / Q000378:metabolism*; D016501:Neurites / Q000187:drug effects* / Q000502:physiology*; D011804:Quinolines / Q000494:pharmacology; D051381:Rats; D012583:Schwann Cells / Q000166:cytology* / Q000378:metabolism*; D012584:Sciatic Nerve / Q000166:cytology; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D013997:Time Factors
chemical_list: C505996:6-amino-4-phenyltetrahydroquinoline; D004791:Enzyme Inhibitors; D007797:Laminin; D007976:Leupeptins; D016328:NF-kappa B; D011804:Quinolines; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde