Analysis of brain region-specific mRNA synthesis and stability by utilizing adult mouse brain slice culture
Summary: Utilization of live animals for mechanistic study is challenging yet pivotal to elucidate pathogenesis of neurological diseases. Here, we present a protocol that employs cultured brain slices derived from adult mice to examine mRNA metabolism. We describe the preparation of acute brain slic...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-06-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166722002295 |
| _version_ | 1818047171497295872 |
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| author | Volodymyr Dzhala Alan J. Fowler Britt A. DiMarzio Kevin J. Staley Jaehong Suh |
| author_facet | Volodymyr Dzhala Alan J. Fowler Britt A. DiMarzio Kevin J. Staley Jaehong Suh |
| author_sort | Volodymyr Dzhala |
| collection | DOAJ |
| description | Summary: Utilization of live animals for mechanistic study is challenging yet pivotal to elucidate pathogenesis of neurological diseases. Here, we present a protocol that employs cultured brain slices derived from adult mice to examine mRNA metabolism. We describe the preparation of acute brain slices and the treatments of RNA synthesis inhibitor and nucleotide analog to examine the effects of ataxin-1 loss-of-function on Bace1 mRNA stability and transcription in cortex. This protocol also includes electrophysiological recording of spontaneous neuronal activity in hippocampus.For complete details on the use and execution of this protocol, please refer to Suh et al. (2019). |
| first_indexed | 2024-12-10T10:01:33Z |
| format | Article |
| id | doaj.art-f7718a9ccfe74cd9b395e6317b2bc707 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-12-10T10:01:33Z |
| publishDate | 2022-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-f7718a9ccfe74cd9b395e6317b2bc7072022-12-22T01:53:21ZengElsevierSTAR Protocols2666-16672022-06-0132101349Analysis of brain region-specific mRNA synthesis and stability by utilizing adult mouse brain slice cultureVolodymyr Dzhala0Alan J. Fowler1Britt A. DiMarzio2Kevin J. Staley3Jaehong Suh4Department of Neurology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02129, USA; Corresponding authorDepartment of Neurology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02129, USA; Genetics and Aging Research Unit, MassGeneral Institute of Neurodegenerative Disease, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02129, USADepartment of Neurology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02129, USA; Genetics and Aging Research Unit, MassGeneral Institute of Neurodegenerative Disease, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02129, USADepartment of Neurology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02129, USADepartment of Neurology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02129, USA; Genetics and Aging Research Unit, MassGeneral Institute of Neurodegenerative Disease, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02129, USA; Corresponding authorSummary: Utilization of live animals for mechanistic study is challenging yet pivotal to elucidate pathogenesis of neurological diseases. Here, we present a protocol that employs cultured brain slices derived from adult mice to examine mRNA metabolism. We describe the preparation of acute brain slices and the treatments of RNA synthesis inhibitor and nucleotide analog to examine the effects of ataxin-1 loss-of-function on Bace1 mRNA stability and transcription in cortex. This protocol also includes electrophysiological recording of spontaneous neuronal activity in hippocampus.For complete details on the use and execution of this protocol, please refer to Suh et al. (2019).http://www.sciencedirect.com/science/article/pii/S2666166722002295Cell BiologyGeneticsGene ExpressionNeuroscience |
| spellingShingle | Volodymyr Dzhala Alan J. Fowler Britt A. DiMarzio Kevin J. Staley Jaehong Suh Analysis of brain region-specific mRNA synthesis and stability by utilizing adult mouse brain slice culture STAR Protocols Cell Biology Genetics Gene Expression Neuroscience |
| title | Analysis of brain region-specific mRNA synthesis and stability by utilizing adult mouse brain slice culture |
| title_full | Analysis of brain region-specific mRNA synthesis and stability by utilizing adult mouse brain slice culture |
| title_fullStr | Analysis of brain region-specific mRNA synthesis and stability by utilizing adult mouse brain slice culture |
| title_full_unstemmed | Analysis of brain region-specific mRNA synthesis and stability by utilizing adult mouse brain slice culture |
| title_short | Analysis of brain region-specific mRNA synthesis and stability by utilizing adult mouse brain slice culture |
| title_sort | analysis of brain region specific mrna synthesis and stability by utilizing adult mouse brain slice culture |
| topic | Cell Biology Genetics Gene Expression Neuroscience |
| url | http://www.sciencedirect.com/science/article/pii/S2666166722002295 |
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