RT Journal Article
SR Electronic
T1 Integrin-linked kinase is a key signal factor involved in Nogo-66-induced inhibition of neurite outgrowth
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.04.14.040444
DO 10.1101/2020.04.14.040444
A1 Ya-ping Yu
A1 Qiang-ping Wang
A1 Jian-Ying Shen
A1 Nan-xiang Xiong
A1 Hua Yu
A1 Peng Fu
A1 Lei Wang
A1 Ye Yuan
A1 Hong-yang Zhao
A1 Fang-cheng Zhang
A1 Hendrik Pool
YR 2020
UL http://biorxiv.org/content/early/2020/04/14/2020.04.14.040444.abstract
AB Nogo-66, the extracellular domain of Nogo-A, has been identified as the most important myelin-associated neuronal growth inhibitor. Evidence suggested that Nogo-66 exert its neurite inhibition effect via a Nogo-66/Protein kinase B (PKB)/Glycogen synthase kinase-3β (GSK-3β)/tau signaling pathway. Integrin-linked kinase (ILK) is a serine/threonine kinase mediating axon upstream growth of PKB and GSK-3β. However, the contribution of ILK to the Nogo-66-induced inhibition of neurite, is not clear. In this study, we set out to reveal the role of ILK on Nogo-66 signaling in vitro and in vivo. To deteremine this directly, Recombinant adenoviruses were constructed to upregulate or downregulate the expresioon of ILK in Neuro 2a (N2a)and analysis the change of downstream molecule and neurite length. The results showed that Nogo-66 inhibited the phosphorylation of ILK, while ILK regulated the phosphorylation of PKB and GSK-3β, and the expression of tau in Nogo-66-treated N2a cells. ILK overexpression through lentivirus vector transfection reduced the inhibitory effect of neurite outgrowth induced by Nogo-66 in cortical neurons. The Tau expression in the complete spinal cord transection rat model was promoted by the overexpression of ILK. Our findings indicated that ILK is a key signal factor involved in Nogo-66-induced inhibition of neurite outgrowth. The mechanism of Nogo-66 signaling pathway was further explained and a proper target for the promotion of neural regeneration was also provided by this study.Competing Interest StatementThe authors have declared no competing interest.Nogo-Aneurite outgrowth inhibitorPKBProtein kinase BGSK-3βGlycogen synthase kinase-3βILKIntegrin-linked kinaseN2aNeuro 2aCNScentral nervous systemOMgpoligodendrocyte-myelin glycoproteinMAGmyelin-associated glycoproteinCSPGchondroitin sulfate proteoglycanNgR1Nogo-66 receptor 1GADPHglyceraldehyde-3-phosphate dehydrogenaseSDSprague-DawleyDMEMDulbecco’s modified Eagle’s mediumPCRpolymerase chain reactionSDS-PAGEsodium dodecyl sulfate-polyacrylamide gel electrophoresisDMSOdimetilsulfóxidoPVDFpolyvinylidene difluoridePBSpolybutylene succinateSPSSThe Statistical Package for the Social SciencesSDstandard deviationANOVAOne-way analysis of variance