Interaction between SDF1 and CXCR4 Promotes Photoreceptor Differentiation via Upregulation of NFκB Pathway Signaling Activity in Pax6 Gene-Transfected Photoreceptor Precursors.
Interaction between SDF1 and CXCR4 Promotes Photoreceptor Differentiation via Upregulation of NFκB Pathway Signaling Activity in Pax6 Gene-Transfected Photoreceptor Precursors.
复制标题
SDF1 和 CXCR4 之间的相互作用通过上调 Pax6 基因转染的光感受器前体中 NFκB 通路信号活性来促进光感受器分化。
DOI：
10.1159/000503929
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发表时间：
2020
期刊：
影响因子：
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通讯作者：
Suzuki N.
中科院分区：
文献类型：
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作者：
Shimizu J;Suzuki T;Hirotsu C;Ueno H;Takada E;Arimitsu N;Ueda Y;Wakisaka S;Suzuki N.
背景CCL 2（也称为单核细胞趋化蛋白1）和CX3CR 1（也称为Fractalkine受体）缺陷小鼠有受损的photoreceptors. ObjectiveWe检查SDF 1和CXCR 4的相互作用对视网膜祖细胞分化为视紫红质阳性photoreceptors.MethodsCloned retinal progenitor通过Pax6基因转染小鼠iPS细胞，然后连续稀释获得。通过巢蛋白、Musashi1、Six3和Chx10 mRNA的表达选择克隆。流式细胞术分析细胞表面蛋白表达。采用实时荧光定量PCR和免疫组化方法检测细胞内mRNA和蛋白水平。视网膜祖细胞的瞬时转染实验进行使用人视紫红质启动子荧光素酶plasmid.ResultsWe选择了10个克隆，表达Six3，Chx10，Crx，Rx1，Nrl，CD73，和视紫红质mRNA，其中，除了视紫红质，感光前体标记。克隆在细胞表面表达CD73和CXCR4，并分化为视紫红质阳性光感受器，通过添加外源性SDF 1来加强。CXCR4抑制剂AMD3100阻断了SDF 1介导的祖细胞向感光细胞的分化。SDF 1可能通过NFκB途径增强人视紫红质启动子的转录活性。向细胞培养物中加入SDF 1可诱导视网膜祖细胞克隆上NFκB的核转位。新生和新生小鼠视网膜表达SDF 1和CXCR 4。光感受器所在的外核层细胞在P14和P56表达CXCR4。结论SDF 1和CXCR 4通过上调NFκB B的活性，部分调控视网膜前体细胞的分化。
BackgroundCCL2 (also known as monocyte chemoattractant protein 1) and CX3CR1 (also known as Fractalkine receptor)-deficient mice have damaged photoreceptors.ObjectivesWe examined the interaction of SDF1 and CXCR4 on the differentiation of retinal progenitors into rhodopsin-positive photoreceptors.MethodsCloned retinal progenitors were obtained by Pax6 gene transfection of mouse iPS cells followed by serial dilution. Clones were selected by expression of nestin, Musashi1, Six3, and Chx10 mRNA. Cell surface protein expression was analyzed by flow cytometry. The levels of mRNA and intracellular protein were examined by real-time PCR and immunochemistry, respectively. Transient transfection experiments of retinal progenitors were conducted using a human rhodopsin promoter luciferase plasmid.ResultsWe selected 10 clones that expressed Six3, Chx10, Crx, Rx1, Nrl, CD73, and rhodopsin mRNA, which, except for rhodopsin, are photoreceptor precursor markers. Clones expressed both CD73 and CXCR4 on the cell surface and differentiated into rhodopsin-positive photoreceptors, which was reinforced by the addition of exogenous SDF1. A CXCR4 inhibitor AMD3100 blocked SDF1-mediated differentiation of progenitors into photoreceptors. SDF1 enhanced human rhodopsin promoter transcription activity, possibly via the NFκB pathway. Addition of SDF1 to the cell culture induced nuclear translocation of NFκB on retinal progenitor cell clones. Neonatal and newborn mouse retinas expressed SDF1 and CXCR4. Cells in the outer nuclear layer where photoreceptors are located expressed CXCR4 at P14 and P56. Cells in the inner nuclear layer expressed SDF1.ConclusionsThese findings suggest that retinal progenitor cell differentiation was at least partly regulated by SDF1 and CXCR4 via upregulation of NFκB activity.