Integrative Analysis of Single-Cell and Bulk Sequencing Data Depicting the Expression and Function of P2ry12 in Microglia Post Ischemia–Reperfusion Injury
<i>P2ry12</i> is a microglial marker gene. Recently, increasing evidence has demonstrated that its expression levels can vary in response to different CNS disorders and can affect microglial functions, such as polarization, plasticity, and migration. However, the expression and function...
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MDPI AG
2023-04-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/7/6772 |
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| author | Chenglong Wang Li Peng Yuan Wang Ying Xue Tianyi Chen Yanyan Ji Yishan Li Yong Zhao Shanshan Yu |
| author_facet | Chenglong Wang Li Peng Yuan Wang Ying Xue Tianyi Chen Yanyan Ji Yishan Li Yong Zhao Shanshan Yu |
| author_sort | Chenglong Wang |
| collection | DOAJ |
| description | <i>P2ry12</i> is a microglial marker gene. Recently, increasing evidence has demonstrated that its expression levels can vary in response to different CNS disorders and can affect microglial functions, such as polarization, plasticity, and migration. However, the expression and function of <i>P2ry12</i> in microglia during ischemia–reperfusion injury (IRI) remain unclear. Here, we developed a computational method to obtain microglia-specific <i>P2ry12</i> genes (MSPGs) using sequencing data associated with IRI. We evaluated the change in comprehensive expression levels of MSPGs during IRI and compared it to the expression of <i>P2ry12</i> to determine similarity. Subsequently, the MSPGs were used to explore the <i>P2ry12</i> functions in microglia through bioinformatics. Moreover, several animal experiments were also conducted to confirm the reliability of the results. The expression of <i>P2ry12</i> was observed to decrease gradually within 24 h post injury. In response, microglia with reduced <i>P2ry12</i> expression showed an increase in the expression of one receptor-encoding gene (<i>Flt1</i>) and three ligand-encoding genes (<i>Nampt</i>, <i>Igf1</i>, and <i>Cxcl2</i>). Furthermore, double-labeling immunofluorescence staining revealed that inhibition of P2ry12 blocked microglial migration towards vessels during IRI. Overall, we employ a combined computational and experimental approach to successfully explore <i>P2ry12</i> expression and function in microglia during IRI. |
| first_indexed | 2024-03-11T05:33:45Z |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T05:33:45Z |
| publishDate | 2023-04-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-528d34d7af3e44ec86bec02772b5b8ea2023-11-17T16:55:50ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-04-01247677210.3390/ijms24076772Integrative Analysis of Single-Cell and Bulk Sequencing Data Depicting the Expression and Function of P2ry12 in Microglia Post Ischemia–Reperfusion InjuryChenglong Wang0Li Peng1Yuan Wang2Ying Xue3Tianyi Chen4Yanyan Ji5Yishan Li6Yong Zhao7Shanshan Yu8Department of Pathology, College of Basic Medicine, Chongqing Medical University, Chongqing 400016, ChinaDepartment of Pathology, College of Basic Medicine, Chongqing Medical University, Chongqing 400016, ChinaDepartment of Pathology, College of Basic Medicine, Chongqing Medical University, Chongqing 400016, ChinaDepartment of Pathology, College of Basic Medicine, Chongqing Medical University, Chongqing 400016, ChinaDepartment of Pathology, College of Basic Medicine, Chongqing Medical University, Chongqing 400016, ChinaDepartment of Pathology, College of Basic Medicine, Chongqing Medical University, Chongqing 400016, ChinaDepartment of Pathology, College of Basic Medicine, Chongqing Medical University, Chongqing 400016, ChinaDepartment of Pathology, College of Basic Medicine, Chongqing Medical University, Chongqing 400016, ChinaDepartment of Pathology, College of Basic Medicine, Chongqing Medical University, Chongqing 400016, China<i>P2ry12</i> is a microglial marker gene. Recently, increasing evidence has demonstrated that its expression levels can vary in response to different CNS disorders and can affect microglial functions, such as polarization, plasticity, and migration. However, the expression and function of <i>P2ry12</i> in microglia during ischemia–reperfusion injury (IRI) remain unclear. Here, we developed a computational method to obtain microglia-specific <i>P2ry12</i> genes (MSPGs) using sequencing data associated with IRI. We evaluated the change in comprehensive expression levels of MSPGs during IRI and compared it to the expression of <i>P2ry12</i> to determine similarity. Subsequently, the MSPGs were used to explore the <i>P2ry12</i> functions in microglia through bioinformatics. Moreover, several animal experiments were also conducted to confirm the reliability of the results. The expression of <i>P2ry12</i> was observed to decrease gradually within 24 h post injury. In response, microglia with reduced <i>P2ry12</i> expression showed an increase in the expression of one receptor-encoding gene (<i>Flt1</i>) and three ligand-encoding genes (<i>Nampt</i>, <i>Igf1</i>, and <i>Cxcl2</i>). Furthermore, double-labeling immunofluorescence staining revealed that inhibition of P2ry12 blocked microglial migration towards vessels during IRI. Overall, we employ a combined computational and experimental approach to successfully explore <i>P2ry12</i> expression and function in microglia during IRI.https://www.mdpi.com/1422-0067/24/7/6772<i>P2ry12</i>microgliaischemia–reperfusion injurybioinformatics |
| spellingShingle | Chenglong Wang Li Peng Yuan Wang Ying Xue Tianyi Chen Yanyan Ji Yishan Li Yong Zhao Shanshan Yu Integrative Analysis of Single-Cell and Bulk Sequencing Data Depicting the Expression and Function of P2ry12 in Microglia Post Ischemia–Reperfusion Injury International Journal of Molecular Sciences <i>P2ry12</i> microglia ischemia–reperfusion injury bioinformatics |
| title | Integrative Analysis of Single-Cell and Bulk Sequencing Data Depicting the Expression and Function of P2ry12 in Microglia Post Ischemia–Reperfusion Injury |
| title_full | Integrative Analysis of Single-Cell and Bulk Sequencing Data Depicting the Expression and Function of P2ry12 in Microglia Post Ischemia–Reperfusion Injury |
| title_fullStr | Integrative Analysis of Single-Cell and Bulk Sequencing Data Depicting the Expression and Function of P2ry12 in Microglia Post Ischemia–Reperfusion Injury |
| title_full_unstemmed | Integrative Analysis of Single-Cell and Bulk Sequencing Data Depicting the Expression and Function of P2ry12 in Microglia Post Ischemia–Reperfusion Injury |
| title_short | Integrative Analysis of Single-Cell and Bulk Sequencing Data Depicting the Expression and Function of P2ry12 in Microglia Post Ischemia–Reperfusion Injury |
| title_sort | integrative analysis of single cell and bulk sequencing data depicting the expression and function of p2ry12 in microglia post ischemia reperfusion injury |
| topic | <i>P2ry12</i> microglia ischemia–reperfusion injury bioinformatics |
| url | https://www.mdpi.com/1422-0067/24/7/6772 |
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