Analysis of reactive astrogliosis in mouse brain using in situ hybridization combined with immunohistochemistry
Summary: Reactive astrogliosis is characterized by a profound change in astrocyte phenotype in response to all CNS injuries. Here, we present a revised in situ hybridization and immunohistochemistry (IHC) protocol to label the reactive astrocytes in the mouse brain. Several approaches for quantifyin...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2021-03-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166721000824 |
| _version_ | 1818728712201306112 |
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| author | Ranjithmenon Muraleedharan Diana Nardini Ronald Raymond Waclaw Biplab Dasgupta |
| author_facet | Ranjithmenon Muraleedharan Diana Nardini Ronald Raymond Waclaw Biplab Dasgupta |
| author_sort | Ranjithmenon Muraleedharan |
| collection | DOAJ |
| description | Summary: Reactive astrogliosis is characterized by a profound change in astrocyte phenotype in response to all CNS injuries. Here, we present a revised in situ hybridization and immunohistochemistry (IHC) protocol to label the reactive astrocytes in the mouse brain. Several approaches for quantifying astrocyte reactivity lacked sensitivity to discriminate across the spectrum. We optimized in situ hybridization followed by IHC. We provide a staining protocol for quantitative measures of astrocyte reactivity as an independent confirmation of the magnitude of reactive gliosis.For complete details on the use and execution of this protocol, please refer to Muraleedharan et al. (2020). |
| first_indexed | 2024-12-17T22:34:21Z |
| format | Article |
| id | doaj.art-ad1f1c487d9449a9b2ece30d1f55ce4d |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-12-17T22:34:21Z |
| publishDate | 2021-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-ad1f1c487d9449a9b2ece30d1f55ce4d2022-12-21T21:30:07ZengElsevierSTAR Protocols2666-16672021-03-0121100375Analysis of reactive astrogliosis in mouse brain using in situ hybridization combined with immunohistochemistryRanjithmenon Muraleedharan0Diana Nardini1Ronald Raymond Waclaw2Biplab Dasgupta3Division of Oncology, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH 45229, USADivision of Experimental Hematology and Cancer Biology, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH 45229, USA; Department of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, OH 45229, USADivision of Experimental Hematology and Cancer Biology, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH 45229, USA; Department of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, OH 45229, USA; Corresponding authorDivision of Oncology, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH 45229, USA; Department of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, OH 45229, USA; Corresponding authorSummary: Reactive astrogliosis is characterized by a profound change in astrocyte phenotype in response to all CNS injuries. Here, we present a revised in situ hybridization and immunohistochemistry (IHC) protocol to label the reactive astrocytes in the mouse brain. Several approaches for quantifying astrocyte reactivity lacked sensitivity to discriminate across the spectrum. We optimized in situ hybridization followed by IHC. We provide a staining protocol for quantitative measures of astrocyte reactivity as an independent confirmation of the magnitude of reactive gliosis.For complete details on the use and execution of this protocol, please refer to Muraleedharan et al. (2020).http://www.sciencedirect.com/science/article/pii/S2666166721000824CancerMetabolismMicroscopyModel organismsIn situ hybridizationNeuroscience |
| spellingShingle | Ranjithmenon Muraleedharan Diana Nardini Ronald Raymond Waclaw Biplab Dasgupta Analysis of reactive astrogliosis in mouse brain using in situ hybridization combined with immunohistochemistry STAR Protocols Cancer Metabolism Microscopy Model organisms In situ hybridization Neuroscience |
| title | Analysis of reactive astrogliosis in mouse brain using in situ hybridization combined with immunohistochemistry |
| title_full | Analysis of reactive astrogliosis in mouse brain using in situ hybridization combined with immunohistochemistry |
| title_fullStr | Analysis of reactive astrogliosis in mouse brain using in situ hybridization combined with immunohistochemistry |
| title_full_unstemmed | Analysis of reactive astrogliosis in mouse brain using in situ hybridization combined with immunohistochemistry |
| title_short | Analysis of reactive astrogliosis in mouse brain using in situ hybridization combined with immunohistochemistry |
| title_sort | analysis of reactive astrogliosis in mouse brain using in situ hybridization combined with immunohistochemistry |
| topic | Cancer Metabolism Microscopy Model organisms In situ hybridization Neuroscience |
| url | http://www.sciencedirect.com/science/article/pii/S2666166721000824 |
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