Protocol to visualize three distinct neuronal ensembles encoding different events in the mouse brain using genetic and viral approaches
Activity tagging of neuronal ensembles has become an important tool in neuroscience. Here, we present a protocol for visualizing separate neuronal ensembles active during three distinct phases of a memory in transgenic mice. We describe steps to label active neurons using viral microinjection, inducing GFP expression under the robust activity marker (RAM) promoter, and transgenic mice, inducing tdTomato (TdT) expression, and immunohistochemical (IHC) visualization of endogenous cFos expression. We then detail procedures for preparation of tissue, imaging, and quantification of memory events. For complete details on the use and execution of this protocol, please refer to Lesuis et al.<sup>1</sup>.
Authors
- Aijm Van Der Hoorn
- Sungmo Park
- Asim J. Rashid (ORCID: https://orcid.org/0000-0002-7802-7620)
- Sylvie L. Lesuis (ORCID: https://orcid.org/0000-0001-7727-9271)
- Andrew J. Mocle (ORCID: https://orcid.org/0009-0007-5146-2730)
- Sheena A. Josselyn (ORCID: https://orcid.org/0000-0001-5451-489X)
- Paul W. Frankland (ORCID: https://orcid.org/0000-0002-1395-3586)
- Matthew N. Hill (ORCID: https://orcid.org/0000-0001-7144-9209)
- Antonietta DeCristofaro
- Angelica M Torelli
Institutions
- University of Calgary (CA)
- Hospital for Sick Children (CA)
Publication Details
- Journal
- STAR Protocols
- Published
- 2025-04-08
- DOI
- https://doi.org/10.1016/j.xpro.2025.103747
- Citations
- 1
- Primary Topic
- Neuroscience and Neuropharmacology Research
- Type
- article
- Field-Weighted Citation Impact
- 0.91
Funders
- Fondation Brain Canada
- ZonMw
- Nederlandse Organisatie voor Wetenschappelijk Onderzoek
- Canadian Institutes of Health Research
- National Institute of Mental Health