SOX2/DRD2信号通路促进脑缺血小鼠星形胶质细胞去分化
SOX2/DRD2 signaling pathway facilitates astrocytic dedifferentiation in cerebral ischemic mice
易旭阳 1康恩铭 1王砚锦 2张坤 1林伟 3武胜昔 1王亚周1
作者信息
- 1. 空军军医大学基础医学院神经生物学教研室,西安 710032
- 2. 空军军医大学基础医学院学员五大队,西安 710032
- 3. 空军军医大学西京医院神经外科,西安 710032
- 折叠
摘要
目的:探讨基于SOX2在神经干细胞(NSCs)-星形胶质细胞中差异性调节的基因多巴胺受体D2(DRD2)在星形胶质细胞去分化中的作用.方法:采用SOX2-GFP小鼠、免疫荧光染色检测SOX2阳性细胞在皮质发育中的谱系变化;采用原代神经干细胞/星形胶质细胞培养、染色质共沉淀测序(ChIP-seq)和Western Blot检测、验证候选基因的表达;采用药理学操控、神经球形成实验、光化学致缺血、免疫荧光染色和行为学评估DRD2信号、星形胶质细胞去分化和动物运动功能.结果:免疫荧光染色结果表明在正常皮质发育过程中,SOX2的表达呈现由神经干细胞向星形胶质细胞的转变.ChIP-seq显示神经干细胞中SOX2结合的增强子和星形胶质细胞中SOX2结合的启动子区均含有DRD2信号相关分子.Western Blot和免疫荧光染色确证了DRD2在神经干细胞和反应性星形胶质细胞的表达.给予DRD2的激动剂喹那环内酯盐酸盐(QH)可在体外、体内促进星形胶质细胞的去分化.并且,QH处理可以提升皮质缺血后的运动功能恢复.结论:激活DRD2信号有助于促进星形胶质细胞的去分化.本研究为未来治疗缺血性卒中提供了一个新思路.
Abstract
Objective:To explore the effects of dopamine receptor D2(DRD2)on astrocytic dedifferentiation based on SOX2-regulated genes in neural stem cells(NSCs)and astrocytes.Methods:Immunofluorescence staining and SOX2-GFP mice were used to examine the lineage differentiation of SOX2-positive cells during the development of cere-bral cortex.Primary NSCs/astrocytes culture,ChIP-seq and Western Blot were adopted to analyze and verify the expres-sion of candidate genes.Pharmacological manipulation,neurosphere formation,photochemical ischemia,immunofluo-rescence staining and behavior tests were adopted to evaluate the effects of activating DRD2 signaling on astrocytic dedif-ferentiation.Results:Immunofluorescence staining demonstrated the NSC-astrocyte switch of SOX2-expression in the normal development of cerebral cortex.ChIP-seq revealed enrichment of DRD2 signaling by SOX2-bound enhancers in NSCs and SOX2-bound promoters in astrocytes.Western Blot and immunofluorescence staining verified the expression of DRD2 in NSCs and reactive astrocytes.Application of quinagolide hydrocholoride(QH),an agonist of DRD2,signifi-cantly promoted astrocytic dedifferentiation both in vitro and in vivo following ischemia.In addition,quinagolide hydro-choloride treatment improved locomotion recovery.Conclusion:Activating DRD2 signaling facilitates astrocytic dedif-ferentiation and may be used to treat ischemic stroke.
关键词
皮质缺血/星形胶质细胞/去分化/SOX2/多巴胺D2受体/小鼠Key words
cerebral ischemia/astrocyte/dedifferentiation/SOX2/dopamine D2 receptor(DRD2)/mouse引用本文复制引用
基金项目
国家自然科学基金(82221001)
国家自然科学基金(82371402)
陕西省自然科学基金(2021JZ-34)
出版年
2024
NETL
NSTL
万方数据