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

Author(s)

Nakahara, Thiago Seike ; Carvalho, Vinicius Miessler de Andrade ; Souza, Mateus Augusto de Andrade ; Trintinalia, Guilherme Ziegler ; Papes, Fabio

• Document type: Journal article
• Source: STAR PROTOCOLS; v. 1, n. 3, p. 27-pg., 2020-12-18.

Abstract

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). (AU)

• FAPESP's process: 12/01689-0 - Molecular study of the epigenome involved in the regulation of olfactory receptor gene expression
• Grantee: Vinicius Miessler de Andrade Carvalho
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• FAPESP's process: 13/03372-6 - Molecular and functional investigation of Polycomb Group proteins and their involvement in olfactory neurogenesis
• Grantee: Mateus Augusto de Andrade Souza
• Support Opportunities: Scholarships in Brazil - Master
• FAPESP's process: 15/50371-0 - Mechanisms underkying how social experience alters hormone levels, sensory signaling and behavior
• Grantee: Fabio Papes
• Support Opportunities: Regular Research Grants
• FAPESP's process: 09/00473-0 - Molecular biology of the olfactory system in mammals: study on the detection of odors and their neural representation in the brain
• Grantee: Fabio Papes
• Support Opportunities: Research Grants - Young Investigators Grants
• FAPESP's process: 14/25594-3 - Optogenetic manipulation of olfactory circuits
• Grantee: Vinicius Miessler de Andrade Carvalho
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate (Direct)
• FAPESP's process: 12/21786-0 - Ascertainment and analysis of olfactory maps in the brain
• Grantee: Vinicius Miessler de Andrade Carvalho
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• FAPESP's process: 12/04026-1 - Functional and Molecular Characterization of OR class receptors expressed in the mammalian Vomeronasal Organ.
• Grantee: Thiago Seike Picoreti Nakahara
• Support Opportunities: Scholarships in Brazil - Master