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Xiao X , Putatunda R , Zhang Y , Soni PV , Li F , Zhang T , Xin M , Luo JJ , Bethea JR , Cheng Y , Hu W
Lymphotoxin beta receptor-mediated NFkappaB signaling promotes glial lineage differentiation and inhibits neuronal lineage differentiation in mouse brain neural stem/progenitor cells
J Neuroinflammation. 2018 Feb 20;15(1) :49
PMID: 29463313    PMCID: PMC5819232    URL: https://www.ncbi.nlm.nih.gov/pubmed/29463313
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BACKGROUND: Lymphotoxin (LT) is a lymphokine mainly expressed in lymphocytes. LTalpha binds one or two membrane-associated LTbeta to form LTalpha2beta1 or LTalpha1beta2 heterotrimers. The predominant LTalpha1beta2 binds to LTbeta receptor (LTbetaR) primarily expressed in epithelial and stromal cells. Most studies on LTbetaR signaling have focused on the organization, development, and maintenance of lymphoid tissues. However, the roles of LTbetaR signaling in the nervous system, particularly in neurogenesis, remain unknown. Here, we investigated the role of LTbetaR-mediated NFkappaB signaling in regulating neural lineage differentiation. METHODS: The C57BL/6J wild-type and GFAP-dnIkappaBalpha transgenic mice were used. Serum-free embryoid bodies were cultured from mouse embryonic stem cells and further induced into neural stem/progenitor cells (NSCs/NPCs). Primary neurospheres were cultured from embryonic and adult mouse brains followed by monolayer culture for amplification/passage. NFkappaB activation was determined by adenovirus-mediated NFkappaB-firefly-luciferase reporter assay and p65/RelB/p52 nuclear translocation assay. LTbetaR mRNA expression was evaluated by quantitative RT-PCR and LTbetaR protein expression was determined by immunohistochemistry and Western blot analysis. Multilabeled immunocytochemistry or immunohistochemistry followed by fluorescent confocal microscopy and quantitative analysis of neural lineage differentiation were performed. Graphing and statistical analysis were performed with GraphPad Prism software. RESULTS: In cultured NSCs/NPCs, LTalpha1beta2 stimulation induced an activation of classical and non-classical NFkappaB signaling. The expression of LTbetaR-like immunoreactivity in GFAP(+)/Sox2(+) NSCs was identified in well-established neurogenic zones of adult mouse brain. Quantitative RT-PCR and Western blot analysis validated the expression of LTbetaR in cultured NSCs/NPCs and brain neurogenic regions. LTbetaR expression was significantly increased during neural induction. LTalpha1beta2 stimulation in cultured NSCs/NPCs promoted astroglial and oligodendrocytic lineage differentiation, but inhibited neuronal lineage differentiation. Astroglial NFkappaB inactivation in GFAP-dnIkappaBalpha transgenic mice rescued LTbetaR-mediated abnormal phenotypes of cultured NSCs/NPCs. CONCLUSION: This study provides the first evidence for the expression and function of LTbetaR signaling in NSCs/NPCs. Activation of LTbetaR signaling promotes glial lineage differentiation. Our results suggest that neurogenesis is regulated by the adaptive immunity and inflammatory responses.
1742-2094 Xiao, Xiao Putatunda, Raj Zhang, Yonggang Soni, Priya V Li, Fang Zhang, Ting Xin, Mingyang Luo, Jin Jun Bethea, John R Cheng, Yuan Hu, Wenhui ORCID: http://orcid.org/0000-0001-8152-6116 DK075964/National Institute of Diabetes and Digestive and Kidney Diseases R01-MH101041/National Institute of Mental Health Journal Article England J Neuroinflammation. 2018 Feb 20;15(1):49. doi: 10.1186/s12974-018-1074-z.