Effect of recombination signal binding protein for immunoglobulin Kappa J region on the proliferation and differentiation of&nbsp; CD133-positive ependymal cells

YE Xin; BIAN Wei; LI Meng-Yi; LOU Yin-Yu; YU Xin-Yue; QI Shuang-Shuang; ZHOU Peng; CUI Huai-Rui; SUN Chen-You

doi:10.16098/j.issn.0529-1356.2022.02.002

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Acta Anatomica Sinica ›› 2022, Vol. 53 ›› Issue (2) : 144-154. DOI: 10.16098/j.issn.0529-1356.2022.02.002

Effect of recombination signal binding protein for immunoglobulin Kappa J region on the proliferation and differentiation of CD133-positive ependymal cells

YE Xin^1,2 BIAN Wei^1,2 LI Meng-yi^1,2 WU An-ting^1,2 ZHANG Ting³ LI Jun-wei^1,2 ZHOU Peng^1,2 CUI Huai-rui¹ SUN Chen-you^1,2*

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Abstract

Objective To explore the effect of recombination signal binding protein for immunoglobulin Kappa J region (RBP-Jκ) on the proliferation and differentiation of CD133-positive ependymal cells and its possible mechanism. Methods RBP-Jκ in CD133-positive ependymal cells of lateral ventricle was interfered with siRNA in the fetuses of embryos which were isolated from 12-day pregnant Institute of Cancer Research(ICR) mouse (3 mice) and knocked out in CD133-CreERTM: :ROSA26-LacZ: :RBP-Jκflox/floxmouse(3 mice) treated with tamoxifen (TAM) intraperitoneally. The proliferation and differentiation of CD133-positive ependymal cells, as well as the related molecules in RBP-Jκ upstream or downstream were evaluated and measured by immunofluorescent staining, Real-time PCR and Western blotting. ResultsBy interfering or knocking out RBP-Jκ, the number of CD133 or β-galactosidase(β-GAL)/doublecortin(DCX) β-tubulin Ⅲ or CD133 β-GAL/microtubule associated protein-2 (MAP-2) neuronal nuclear antigen(NeuN) or CD133 β-GAL/proliferating cell nuclear antigen(PCNA) double-positive cells were significantly increased in vitro or in vivo. Meanwhile, the result from Real-time PCR and Western blotting showed that RBP-Jκ and hairy and enhancer of split1(Hes1) mRNA and their protein expression levels decreased significantly, while Notch signaling pathway receptor 1(Notch1) mRNA and its protein expression levels increased significantly. Conclusion The interference or knockout of RBP-Jκ will lead to upregulateing Notch1 and downregulate Hes1 expression levels, which eventually increase the proliferation and differentiation of CD133-positive ependymal cells.

Key words

CD133-positive ependymal cell / Notch signaling pathway / Recombination signal binding protein for immunoglobulin Kappa J region / Immunofluorescentce / Mouse

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YE Xin BIAN Wei LI Meng-yi WU An-ting ZHANG Ting LI Jun-wei ZHOU Peng CUI Huai-rui SUN Chen-you. Effect of recombination signal binding protein for immunoglobulin Kappa J region on the proliferation and differentiation of CD133-positive ependymal cells[J]. Acta Anatomica Sinica. 2022, 53(2): 144-154 https://doi.org/10.16098/j.issn.0529-1356.2022.02.002

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