Multimodal mapping of cell types and projections in the central nucleus of the amygdala
- Janelia Research Campus, United States
- German Center for Neurodegenerative Diseases, Germany
- Friedrich Miescher Institute, Switzerland
- Howard Hughes Medical Institute, University of California, San Diego, United States
Abstract
The central nucleus of the amygdala (CEA) is a brain region that integrates external and internal sensory information and executes innate and adaptive behaviors through distinct output pathways. Despite its complex functions, the diversity of molecularly defined neuronal types in the CEA and their contributions to major axonal projection targets have not been examined systematically. Here, we performed single-cell RNA-sequencing (scRNA-Seq) to classify molecularly defined cell types in the CEA and identified marker genes to map the location of these neuronal types using expansion asisted iterative fluorescence in situ hybridization (EASI-FISH). We developed new methods to integrate EASI-FISH with 5-plex retrograde axonal labeling to determine the spatial, morphological, and connectivity properties of ~30,000 molecularly defined CEA neurons. Our study revealed spatio-molecular organization of the CEA, with medial and lateral CEA associated with distinct molecularly defined cell families. We also found a long-range axon projection network from the CEA, where target regions receive inputs from multiple molecularly defined cell types. Axon collateralization was found primarily among projections to hindbrain targets, which are distinct from forebrain projections. This resource reports marker gene combinations for molecularly defined cell types and axon-projection types, which will be useful for selective interrogation of these neuronal populations to study their contributions to the diverse functions of the CEA.
Data availability
All data generated in this study have been deposited at figshare (https://figshare.com/s/a031a8dfca1b4d25d3de). We also provide an interactive data portal for data visualization at http://multifish-data.janelia.org/. The single-cell RNA-seq dataset generated in this study has been deposited to GEO (Gene Expression Omnibus, https://www.ncbi.nlm.nih.gov/geo/) with accession number GSE213828. Any additional information required to reanalyze the data reported in this paper is available from the lead contacts upon request.
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Single cell RNA-sequencing in the mouse central amygdala (CEA)NCBI Gene Expression Omnibus, GSE213828.
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EASI-FISH reveals spatial and axonal projection patterns of molecularly defined cell types in the central amygdala (CEA)Figshare, https://doi.org/10.25378/janelia.21171373.
Article and author information
Author details
Funding
Howard Hughes Medical Institute
- Scott Sternson
Dementia Research Switzerland
- Sabine Krabbe
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Animal experimentation: All methods for animal care and use were conducted according to National Institutes of Health guidelines for animal research and approved by the Institutional Animal Care and Use Committee (IACUC) at Janelia Research Campus (Protocol number: 19-174).
Copyright
© 2023, Wang et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Multimodal mapping of cell types and projections in the central nucleus of the amygdala
eLife 12:e84262.
https://doi.org/10.7554/eLife.84262
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