Cortex-specific transcriptome profiling reveals upregulation of interferon-regulated genes after deeper cerebral hypoperfusion in mice
Background: Chronic cerebral hypoperfusion (CCH) is commonly accompanied by brain injury and glial activation. In addition to white matter lesions, the intensity of CCH greatly affects the degree of gray matter damage. However, little is understood about the underlying molecular mechanisms related t...
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| Format: | Article |
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Frontiers Media S.A.
2023-03-01
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| Series: | Frontiers in Physiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fphys.2023.1056354/full |
| _version_ | 1797884331828445184 |
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| author | Zengyu Zhang Zimin Guo Zhilan Tu Hualan Yang Chao Li Mengting Hu Yuan Zhang Pengpeng Jin Shuangxing Hou |
| author_facet | Zengyu Zhang Zimin Guo Zhilan Tu Hualan Yang Chao Li Mengting Hu Yuan Zhang Pengpeng Jin Shuangxing Hou |
| author_sort | Zengyu Zhang |
| collection | DOAJ |
| description | Background: Chronic cerebral hypoperfusion (CCH) is commonly accompanied by brain injury and glial activation. In addition to white matter lesions, the intensity of CCH greatly affects the degree of gray matter damage. However, little is understood about the underlying molecular mechanisms related to cortical lesions and glial activation following hypoperfusion. Efforts to investigate the relationship between neuropathological alternations and gene expression changes support a role for identifying novel molecular pathways by transcriptomic mechanisms.Methods: Chronic cerebral ischemic injury model was induced by the bilateral carotid artery stenosis (BCAS) using 0.16/0.18 mm microcoils. Cerebral blood flow (CBF) was evaluated using laser speckle contrast imaging (LSCI) system. Spatial learning and memory were assessed by Morris water maze test. Histological changes were evaluated by Hematoxylin staining. Microglial activation and neuronal loss were further examined by immunofluorescence staining. Cortex-specific gene expression profiling analysis was performed in sham and BCAS mice, and then validated by quantitative RT-PCR and immunohistochemistry (IHC).Results: In our study, compared with the sham group, the right hemisphere CBF of BCAS mice decreased to 69% and the cognitive function became impaired at 4 weeks postoperation. Besides, the BCAS mice displayed profound gray matter damage, including atrophy and thinning of the cortex, accompanied by neuronal loss and increased activated microglia. Gene set enrichment analysis (GSEA) revealed that hypoperfusion-induced upregulated genes were significantly enriched in the pathways of interferon (IFN)-regulated signaling along with neuroinflammation signaling. Ingenuity pathway analysis (IPA) predicted the importance of type I IFN signaling in regulating the CCH gene network. The obtained RNA-seq data were validated by qRT-PCR in cerebral cortex, showing consistency with the RNA-seq results. Also, IHC staining revealed elevated expression of IFN-inducible protein in cerebral cortex following BCAS-hypoperfusion.Conclusion: Overall, the activation of IFN-mediated signaling enhanced our understanding of the neuroimmune responses induced by CCH. The upregulation of IFN-regulated genes (IRGs) might exert a critical impact on the progression of cerebral hypoperfusion. Our improved understanding of cortex-specific transcriptional profiles will be helpful to explore potential targets for CCH. |
| first_indexed | 2024-04-10T04:05:50Z |
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| id | doaj.art-4d0a4d6d6b0c48788c19994e956cc633 |
| institution | Directory Open Access Journal |
| issn | 1664-042X |
| language | English |
| last_indexed | 2024-04-10T04:05:50Z |
| publishDate | 2023-03-01 |
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| series | Frontiers in Physiology |
| spelling | doaj.art-4d0a4d6d6b0c48788c19994e956cc6332023-03-13T05:44:27ZengFrontiers Media S.A.Frontiers in Physiology1664-042X2023-03-011410.3389/fphys.2023.10563541056354Cortex-specific transcriptome profiling reveals upregulation of interferon-regulated genes after deeper cerebral hypoperfusion in miceZengyu Zhang0Zimin Guo1Zhilan Tu2Hualan Yang3Chao Li4Mengting Hu5Yuan Zhang6Pengpeng Jin7Shuangxing Hou8Department of Neurology, Shanghai Pudong Hospital, Fudan University, Shanghai, ChinaDepartment of Neurology, Shanghai Pudong Hospital, Fudan University, Shanghai, ChinaDepartment of Neurology, Shanghai Pudong Hospital, Fudan University, Shanghai, ChinaDepartment of Neurology, Shanghai Pudong Hospital, Fudan University, Shanghai, ChinaSchool of Pharmacy, Hubei University of Science and Technology, Hubei, ChinaDepartment of Neurology, Shanghai Pudong Hospital, Fudan University, Shanghai, ChinaDepartment of Vascular Surgery, Shanghai Pudong Hospital, Fudan University, Shanghai, ChinaDepartment of Chronic Disease Management, Shanghai Pudong Hospital, Fudan University, Shanghai, ChinaDepartment of Neurology, Shanghai Pudong Hospital, Fudan University, Shanghai, ChinaBackground: Chronic cerebral hypoperfusion (CCH) is commonly accompanied by brain injury and glial activation. In addition to white matter lesions, the intensity of CCH greatly affects the degree of gray matter damage. However, little is understood about the underlying molecular mechanisms related to cortical lesions and glial activation following hypoperfusion. Efforts to investigate the relationship between neuropathological alternations and gene expression changes support a role for identifying novel molecular pathways by transcriptomic mechanisms.Methods: Chronic cerebral ischemic injury model was induced by the bilateral carotid artery stenosis (BCAS) using 0.16/0.18 mm microcoils. Cerebral blood flow (CBF) was evaluated using laser speckle contrast imaging (LSCI) system. Spatial learning and memory were assessed by Morris water maze test. Histological changes were evaluated by Hematoxylin staining. Microglial activation and neuronal loss were further examined by immunofluorescence staining. Cortex-specific gene expression profiling analysis was performed in sham and BCAS mice, and then validated by quantitative RT-PCR and immunohistochemistry (IHC).Results: In our study, compared with the sham group, the right hemisphere CBF of BCAS mice decreased to 69% and the cognitive function became impaired at 4 weeks postoperation. Besides, the BCAS mice displayed profound gray matter damage, including atrophy and thinning of the cortex, accompanied by neuronal loss and increased activated microglia. Gene set enrichment analysis (GSEA) revealed that hypoperfusion-induced upregulated genes were significantly enriched in the pathways of interferon (IFN)-regulated signaling along with neuroinflammation signaling. Ingenuity pathway analysis (IPA) predicted the importance of type I IFN signaling in regulating the CCH gene network. The obtained RNA-seq data were validated by qRT-PCR in cerebral cortex, showing consistency with the RNA-seq results. Also, IHC staining revealed elevated expression of IFN-inducible protein in cerebral cortex following BCAS-hypoperfusion.Conclusion: Overall, the activation of IFN-mediated signaling enhanced our understanding of the neuroimmune responses induced by CCH. The upregulation of IFN-regulated genes (IRGs) might exert a critical impact on the progression of cerebral hypoperfusion. Our improved understanding of cortex-specific transcriptional profiles will be helpful to explore potential targets for CCH.https://www.frontiersin.org/articles/10.3389/fphys.2023.1056354/fullbilateral carotid artery stenosiscerebral cortexRNA-seqtype I interferonchronic cerebral hypoperfusion (CCH) |
| spellingShingle | Zengyu Zhang Zimin Guo Zhilan Tu Hualan Yang Chao Li Mengting Hu Yuan Zhang Pengpeng Jin Shuangxing Hou Cortex-specific transcriptome profiling reveals upregulation of interferon-regulated genes after deeper cerebral hypoperfusion in mice Frontiers in Physiology bilateral carotid artery stenosis cerebral cortex RNA-seq type I interferon chronic cerebral hypoperfusion (CCH) |
| title | Cortex-specific transcriptome profiling reveals upregulation of interferon-regulated genes after deeper cerebral hypoperfusion in mice |
| title_full | Cortex-specific transcriptome profiling reveals upregulation of interferon-regulated genes after deeper cerebral hypoperfusion in mice |
| title_fullStr | Cortex-specific transcriptome profiling reveals upregulation of interferon-regulated genes after deeper cerebral hypoperfusion in mice |
| title_full_unstemmed | Cortex-specific transcriptome profiling reveals upregulation of interferon-regulated genes after deeper cerebral hypoperfusion in mice |
| title_short | Cortex-specific transcriptome profiling reveals upregulation of interferon-regulated genes after deeper cerebral hypoperfusion in mice |
| title_sort | cortex specific transcriptome profiling reveals upregulation of interferon regulated genes after deeper cerebral hypoperfusion in mice |
| topic | bilateral carotid artery stenosis cerebral cortex RNA-seq type I interferon chronic cerebral hypoperfusion (CCH) |
| url | https://www.frontiersin.org/articles/10.3389/fphys.2023.1056354/full |
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