Abstract
Acute stress leads to sequential activation of functional brain networks. A biologically relevant question is exactly which (single) cells belonging to brain networks are changed in activity over time after acute stress across the entire brain. We developed a preprocessing and analytical pipeline to chart whole-brain immediate early genes’ expression—as proxy for cellular activity—after a single stressful foot shock in four dimensions: that is, from functional networks up to three-dimensional (3D) single-cell resolution and over time. The pipeline is available as an R package. Most brain areas (96%) showed increased numbers of c-fos+ cells after foot shock, yet hypothalamic areas stood out as being most active and prompt in their activation, followed by amygdalar, prefrontal, hippocampal, and finally, thalamic areas. At the cellular level, c-fos+ density clearly shifted over time across subareas, as illustrated for the basolateral amygdala. Moreover, some brain areas showed increased numbers of c-fos+ cells, while others—like the dentate gyrus—dramatically increased c-fos intensity in just a subset of cells, reminiscent of engrams; importantly, this “strategy” changed after foot shock in half of the brain areas. One of the strengths of our approach is that single-cell data were simultaneously examined across all of the 90 brain areas and can be visualized in 3D in our interactive web portal.
Original language | English |
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Article number | e2114002119 |
Pages (from-to) | 1-9 |
Number of pages | 9 |
Journal | Proceedings of the National Academy of Sciences of the United States of America |
Volume | 119 |
Issue number | 8 |
DOIs | |
Publication status | Published - 22 Feb 2022 |
Bibliographical note
Funding Information:ACKNOWLEDGMENTS. We acknowledge the MIND facility (University Medical Center Utrecht Brain Center) for 3D imaging by light-sheet microscopy (https:// mindresearchfacility.nl/) and Roger Koot for the IT infrastructure. We thank Kevin Kemna for suggesting the creation of the R package and the helpful feedback on the analyses. This work was supported by the Consortium of Individual Development and BRAINSCAPES, which are funded through the Gravitation Program of the Dutch Ministry of Education, Culture, and Science and Netherlands Organization for Scientific Research Grants 024.001.003 and 024. 004.012.
Publisher Copyright:
© 2022 National Academy of Sciences. All rights reserved.
Keywords
- Acute stress
- Brain-wide analysis
- c-fos
- Foot shock
- Whole brain