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Transgenic strategies for combinatorial expression of fluorescent proteins in the nervous system
Abstract
Detailed analysis of neuronal network architecture requires the development of new methods. Here we present strategies to visualize synaptic circuits by genetically labelling neurons with multiple, distinct colours. In Brainbow transgenes, Cre/lox recombination is used to create a stochastic choice of expression between three or more fluorescent proteins (XFPs). Integration of tandem Brainbow copies in transgenic mice yielded combinatorial XFP expression, and thus many colours, thereby providing a way to distinguish adjacent neurons and visualize other cellular interactions. As a demonstration, we reconstructed hundreds of neighbouring axons and multiple synaptic contacts in one small volume of a cerebellar lobe exhibiting approximately 90 colours. The expression in some lines also allowed us to map glial territories and follow glial cells and neurons over time in vivo. The ability of the Brainbow system to label uniquely many individual cells within a population may facilitate the analysis of neuronal circuitry on a large scale.
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“We confirmed our hypothesis of a common progenitor pool by lineage tracing the progeny of Wnt1+ progenitors (by crossing Wnt1-CRE2 (Lewis et al., 2013) with R26R-Confetti mice (Livet et al., 2007; Snippert et al., 2010), Figure 2H,I.”
From A cellular and spatial map of the choroid plexus across brain ventricles and ages (2021) · cites this paper as Livet et al., 2007
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Metadata
- Authors
- Livet, J., Weissman, T. A., Kang, H., Draft, R. W., Lu, J., Bennis, R. A., Sanes, J. R., & Lichtman, J. W.
- Affiliations
- Harvard University (US)
- Corresponding author
- Lichtman, Jeff W.
- Venue
- Nature
- Published
- 2007-11
- Volume
- 450
- Issue
- 7166
- Pages
- 56-62
- ISSN
- 0028-0836 (ISSN-L)
- 1476-4687
- OpenAlex
- W2110232607
- Cited by
- 1,876
- References
- 42
- Citations by year
- 2026: 52 · 2025: 76 · 2024: 68 · 2023: 90 · 2022: 88 · 2021: 88 · 2020: 115 · 2019: 104 · 2018: 124 · 2017: 101 · 2016: 117 · 2015: 101 · 2014: 124 · 2013: 161 · 2012: 110
- FWCI
- 30.01
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature (publishedVersion)
- PubMed (publishedVersion)
- RePEc: Research Papers in Economics (submittedVersion)
- Topics
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
- Life Sciences › Neuroscience › Developmental Neuroscience › Neurogenesis and neuroplasticity mechanisms
- Keywords
- Brainbow, transgenic mice, fluorescent proteins, Cre-loxP recombination, stochastic gene expression, neuronal circuit mapping, axon reconstruction, synaptic circuits, cerebellum, hippocampus
- MeSH terms
- Animals, Attachment Sites, Microbiological, Axons, Cell Communication, Cell Line, Cerebellum, Color, Genetic Engineering, Humans, Luminescent Proteins, Mice, Transgenic, Nervous System, Neural Pathways, Neuroglia, Recombination, Genetic, Stochastic Processes, Synapses, Time Factors, Gene Expression, Transgenes, Integrases, Mice
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