Detection of Activated Mouse Neurons with Temporal Resolution via Dual c-Fos Staining

This protocol combines fluorescent in situ hybridization and immunostaining to simultaneously detect, in histological sections from the same animal, subpopulations of neurons activated after two episodes of sensory stimulation. It allows the identification of groups of cells singly activated by eith...

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Bibliographic Details
Published in:STAR protocols 2020-12, Vol.1 (3), p.100153-100153, Article 100153
Main Authors: Nakahara, Thiago Seike, Carvalho, Vinicius Miessler de Andrade, Souza, Mateus Augusto de Andrade, Trintinalia, Guilherme Ziegler, Papes, Fabio
Format: Article
Language:English
Subjects:
Animals
Immunohistochemistry
Mice, Inbred C57BL
Neurons - cytology
Proto-Oncogene Proteins c-fos - metabolism
Protocol
RNA - metabolism
Staining and Labeling
Time Factors
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Summary:This protocol combines fluorescent in situ hybridization and immunostaining to simultaneously detect, in histological sections from the same animal, subpopulations of neurons activated after two episodes of sensory stimulation. It allows the identification of groups of cells singly activated by either stimulus or co-activated by both stimuli. Our method results in nuclear staining for c-Fos mRNA and c-Fos protein, allowing better spatial and temporal resolution than previously published protocols, although it requires quick brain fixation. For complete details on the use and execution of this protocol, please refer to Carvalho et al. (2015, 2020). [Display omitted] •Dual staining identifies activated neurons after two episodes of sensory stimulation•Hybridization phase includes c-Fos mRNA hybridization and signal development•Immunostaining phase includes antibody detection of c-Fos protein•Cells activated by stimuli 1 and 2 are labeled in green and red, respectively This protocol combines fluorescent in situ hybridization and immunostaining to simultaneously detect, in histological sections from the same animal, subpopulations of neurons activated after two episodes of sensory stimulation. It allows the identification of groups of cells singly activated by either stimulus or co-activated by both stimuli. Our method results in nuclear staining for c-Fos mRNA and c-Fos protein, allowing better spatial and temporal resolution than previously published protocols, although it requires quick brain fixation.
ISSN:2666-1667
2666-1667
DOI:10.1016/j.xpro.2020.100153