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Imaging Neuronal Subsets in Transgenic Mice Expressing Multiple Spectral Variants of GFP
Open accessAbstract
We generated transgenic mice in which red, green, yellow, or cyan fluorescent proteins (together termed XFPs) were selectively expressed in neurons. All four XFPs labeled neurons in their entirety, including axons, nerve terminals, dendrites, and dendritic spines. Remarkably, each of 25 independently generated transgenic lines expressed XFP in a unique pattern, even though all incorporated identical regulatory elements (from the thyl gene). For example, all retinal ganglion cells or many cortical neurons were XFP positive in some lines, whereas only a few ganglion cells or only layer 5 cortical pyramids were labeled in others. In some lines, intense labeling of small neuronal subsets provided a Golgi-like vital stain. In double transgenic mice expressing two different XFPs, it was possible to differentially label 3 neuronal subsets in a single animal.
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“Thy1-GFP-M is a transgenic reporter line in which eGFP is expressed in a subset of layer II/III and V cortical neurons (Feng et al., 2000).”
From Dendritic Spine Instability Leads to Progressive Neocortical Spine Loss in a Mouse Model of Huntington's Disease (2013) · cites this paper as Feng et al., 2000
In the citing paper
“So far, the majority of in vivo imaging has been performed using thy1 transgenic lines, which express fluorescent proteins sparsely in a subset of cortical neurons, often with mixed cell populations, including both pyramidal neurons and interneurons [18].”
From Experience-dependent structural plasticity in the cortex (2011) · cites this paper as [18]
In the citing paper
“Currently the only transgenic mouse approach demonstrated to reliably achieve AAV-like high-level expression is the use of the Thy1.2 promoter in randomly integrated transgenes (Arenkiel et al., 2007; Dana et al., 2014; Feng et al., 2000; Zhao et al., 2008), presumably with multiple copies at the insertion site.”
From Transgenic mice for intersectional targeting of neural sensors and effectors with high specificity and performance (2015) · cites this paper as Feng et al., 2000
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“C57BL/6 transgenic mice (H-line) expressing YFP in a subset of layer 5 cortical neurons (Feng et al., 2000) were purchased from The Jackson Laboratory (Bar Harbor, ME) and bred at the University of British Columbia animal facilities.”
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“Illumination was performed using a single red LED (Luxeon K2, 627 nm) and fluorescence filter set as outlined previously (Ferezou et al., 2006) with an Olympus XLfluor 4 0.28 NA objective focused 200 m below the surface and imaged with a Dalsa 1M60 camera using a similar analysis as that used for…”
MethodInfluentialHippocampal ripples down-regulate synapses
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Metadata
- Authors
- Feng, G., Mellor, R. H., Bernstein, M., Keller-Peck, C., Nguyen, Q. T., Wallace, M., Nerbonne, J. M., Lichtman, J. W., & Sanes, J. R.
- Affiliations
- Washington University in St. Louis (US)
- Corresponding author
- Sanes, Joshua R.
- ORCID
- Venue
- Neuron
- Publisher
- Elsevier BV
- Published
- 2000-10
- Volume
- 28
- Issue
- 1
- Pages
- 41-51
- ISSN
- 0896-6273 (ISSN-L)
- OpenAlex
- W2127810309
- Cited by
- 3,042
- References
- 52
- Citations by year
- 2026: 42 · 2025: 102 · 2024: 105 · 2023: 126 · 2022: 128 · 2021: 132 · 2020: 153 · 2019: 133 · 2018: 149 · 2017: 132 · 2016: 164 · 2015: 175 · 2014: 173 · 2013: 180 · 2012: 164
- FWCI
- 25.11
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Neuron (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
- Life Sciences › Neuroscience › Developmental Neuroscience › Neurogenesis and neuroplasticity mechanisms
- Keywords
- transgenic mice, neuronal labeling, retinal ganglion cells, cortical pyramidal neurons, dendritic spines, axons, presynaptic terminals
- MeSH terms
- Animals, Axons, Cerebral Cortex, Color, Dendrites, Light, Luminescent Proteins, Mice, Transgenic, Microscopy, Fluorescence, Neuromuscular Junction, Neurons, Regulatory Sequences, Nucleic Acid, Retinal Ganglion Cells, Synapses, Presynaptic Terminals, Thy-1 Antigens, Cell Lineage, Transgenes, Green Fluorescent Proteins, Mice
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