{"title":"急性缺血性中风的转录组分析:星形胶质细胞和小胶质细胞在卒中后表达转录本的替代剪接方面表现出差异。","authors":"Bingxue Jin, Yilai Han, Fang Xu, Junjie Wang, Yunzhi Zhao, Haijie Liu, Fei Wang, Ze Wang, Wanting Lu, Mingyang Wang, Lili Cui, Yinan Zhao, Junwei Hao, Guoliang Chai","doi":"10.1096/fj.202400341R","DOIUrl":null,"url":null,"abstract":"<p>Astrocytes and microglia undergo dynamic and complex morphological and functional changes following ischemic stroke, which are instrumental in both inflammatory responses and neural repair. While gene expression alterations poststroke have been extensively studied, investigations into posttranscriptional regulatory mechanisms, specifically alternative splicing (AS), remain limited. Utilizing previously reported Ribo-Tag-seq data, this study analyzed AS alterations in poststroke astrocytes and microglia from young adult male and female mice. Our findings reveal that in astrocytes, compared to the sham group, 109 differential alternative splicing (DAS) events were observed at 4 h poststroke, which increased to 320 at day 3. In microglia, these numbers were 316 and 266, respectively. Interestingly, the disparity between DAS genes and differentially expressed genes is substantial, with fewer than 10 genes shared at both poststroke time points in astrocytes and microglia. Gene ontology enrichment analysis revealed the involvement of these DAS genes in diverse functions, encompassing immune response (<i>Adam8</i>, <i>Ccr1</i>), metabolism (<i>Acsl6</i>, <i>Pcyt2, Myo5a</i>), and developmental cell growth (<i>App</i>), among others. Selective DAS events were further validated by semiquantitative RT-PCR. Overall, this study comprehensively describes the AS alterations in astrocytes and microglia during the hyperacute and acute phases of ischemic stroke and underscores the significance of certain hub DAS events in neuroinflammatory processes.</p>","PeriodicalId":50455,"journal":{"name":"FASEB Journal","volume":null,"pages":null},"PeriodicalIF":4.4000,"publicationDate":"2024-08-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1096/fj.202400341R","citationCount":"0","resultStr":"{\"title\":\"Translatome analysis in acute ischemic stroke: Astrocytes and microglia exhibit differences in poststroke alternative splicing of expressed transcripts\",\"authors\":\"Bingxue Jin, Yilai Han, Fang Xu, Junjie Wang, Yunzhi Zhao, Haijie Liu, Fei Wang, Ze Wang, Wanting Lu, Mingyang Wang, Lili Cui, Yinan Zhao, Junwei Hao, Guoliang Chai\",\"doi\":\"10.1096/fj.202400341R\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>Astrocytes and microglia undergo dynamic and complex morphological and functional changes following ischemic stroke, which are instrumental in both inflammatory responses and neural repair. While gene expression alterations poststroke have been extensively studied, investigations into posttranscriptional regulatory mechanisms, specifically alternative splicing (AS), remain limited. Utilizing previously reported Ribo-Tag-seq data, this study analyzed AS alterations in poststroke astrocytes and microglia from young adult male and female mice. Our findings reveal that in astrocytes, compared to the sham group, 109 differential alternative splicing (DAS) events were observed at 4 h poststroke, which increased to 320 at day 3. In microglia, these numbers were 316 and 266, respectively. Interestingly, the disparity between DAS genes and differentially expressed genes is substantial, with fewer than 10 genes shared at both poststroke time points in astrocytes and microglia. Gene ontology enrichment analysis revealed the involvement of these DAS genes in diverse functions, encompassing immune response (<i>Adam8</i>, <i>Ccr1</i>), metabolism (<i>Acsl6</i>, <i>Pcyt2, Myo5a</i>), and developmental cell growth (<i>App</i>), among others. Selective DAS events were further validated by semiquantitative RT-PCR. Overall, this study comprehensively describes the AS alterations in astrocytes and microglia during the hyperacute and acute phases of ischemic stroke and underscores the significance of certain hub DAS events in neuroinflammatory processes.</p>\",\"PeriodicalId\":50455,\"journal\":{\"name\":\"FASEB Journal\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":4.4000,\"publicationDate\":\"2024-08-03\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://onlinelibrary.wiley.com/doi/epdf/10.1096/fj.202400341R\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"FASEB Journal\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1096/fj.202400341R\",\"RegionNum\":2,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"BIOCHEMISTRY & MOLECULAR BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"FASEB Journal","FirstCategoryId":"99","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1096/fj.202400341R","RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
引用次数: 0
摘要
缺血性脑卒中后,星形胶质细胞和小胶质细胞会发生动态而复杂的形态和功能变化,这些变化在炎症反应和神经修复中起着重要作用。虽然对中风后基因表达改变的研究已经非常广泛,但对转录后调控机制,特别是替代剪接(AS)的研究仍然有限。本研究利用之前报道的 Ribo-Tag-seq 数据,分析了年轻成年雄性和雌性小鼠中风后星形胶质细胞和小胶质细胞的 AS 改变。我们的研究结果表明,与假组相比,中风后 4 小时在星形胶质细胞中观察到 109 个差异替代剪接(DAS)事件,到第 3 天时增加到 320 个。在小胶质细胞中,这一数字分别为 316 和 266。有趣的是,DAS基因和差异表达基因之间的差异很大,在中风后的两个时间点,星形胶质细胞和小胶质细胞中共享的基因不到10个。基因本体富集分析显示,这些DAS基因参与了多种功能,包括免疫反应(Adam8、Ccr1)、新陈代谢(Acsl6、Pcyt2、Myo5a)和细胞生长发育(App)等。半定量 RT-PCR 进一步验证了选择性 DAS 事件。总之,本研究全面描述了缺血性脑卒中超急性期和急性期星形胶质细胞和小胶质细胞的 AS 改变,并强调了神经炎症过程中某些枢纽 DAS 事件的重要性。
Translatome analysis in acute ischemic stroke: Astrocytes and microglia exhibit differences in poststroke alternative splicing of expressed transcripts
Astrocytes and microglia undergo dynamic and complex morphological and functional changes following ischemic stroke, which are instrumental in both inflammatory responses and neural repair. While gene expression alterations poststroke have been extensively studied, investigations into posttranscriptional regulatory mechanisms, specifically alternative splicing (AS), remain limited. Utilizing previously reported Ribo-Tag-seq data, this study analyzed AS alterations in poststroke astrocytes and microglia from young adult male and female mice. Our findings reveal that in astrocytes, compared to the sham group, 109 differential alternative splicing (DAS) events were observed at 4 h poststroke, which increased to 320 at day 3. In microglia, these numbers were 316 and 266, respectively. Interestingly, the disparity between DAS genes and differentially expressed genes is substantial, with fewer than 10 genes shared at both poststroke time points in astrocytes and microglia. Gene ontology enrichment analysis revealed the involvement of these DAS genes in diverse functions, encompassing immune response (Adam8, Ccr1), metabolism (Acsl6, Pcyt2, Myo5a), and developmental cell growth (App), among others. Selective DAS events were further validated by semiquantitative RT-PCR. Overall, this study comprehensively describes the AS alterations in astrocytes and microglia during the hyperacute and acute phases of ischemic stroke and underscores the significance of certain hub DAS events in neuroinflammatory processes.
期刊介绍:
The FASEB Journal publishes international, transdisciplinary research covering all fields of biology at every level of organization: atomic, molecular, cell, tissue, organ, organismic and population. While the journal strives to include research that cuts across the biological sciences, it also considers submissions that lie within one field, but may have implications for other fields as well. The journal seeks to publish basic and translational research, but also welcomes reports of pre-clinical and early clinical research. In addition to research, review, and hypothesis submissions, The FASEB Journal also seeks perspectives, commentaries, book reviews, and similar content related to the life sciences in its Up Front section.