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Versatile genetic paintbrushes: Brainbow technologies.

Richier B, Salecker I - Wiley Interdiscip Rev Dev Biol (2014)

Bottom Line: While being continuously refined, Brainbow technologies have thus found a firm place in the genetic toolboxes of developmental and neurobiologists.For further resources related to this article, please visit the WIREs website.The authors have declared no conflicts of interest for this article.

View Article: PubMed Central - PubMed

Affiliation: MRC National Institute for Medical Research, Division of Molecular Neurobiology, London, UK.

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Four examples of Brainbow technologies at work. (a) Purkinje cells in the mouse cerebellum are visualized in seven colors (i–vii) using Brainbow-3.1 and L7-Cre transgenes, as well as antibody amplification. (Reprinted with permission from Ref 16. Copyright 2013 Nature Publishing Group) Scale bar, 20 µm. (b) Pyramidal neurons in the P28 cortex of a CAG-CreERTM mouse are labeled by combinations of co-electroporated MAGIC Cytbow and Nucbow markers at E15. The image was acquired by two-photon microscopy. (Reprinted with permission from Ref 17. Copyright 2014 Elsevier Ltd.) Scale bar, 100 µm. (c) Neurites of lamina and medulla neuron subtypes (ln, mn) in the adult Drosophila optic lobe are visualized by endogenous fluorescent protein signals using a Flybow-2.0B transgene, activated by hs-mFLP5 and NP4151-Gal4—an enhancer trap insertion into the Netrin B locus. The image represents a single optical section. Several neurons (arrowheads) are suitable for tracing in stacks. Photoreceptor axons are visualized by immunolabeling with mAb24B10 (blue). Scale bar, 20 µm. (d) Nuclei of epithelial cell clones in a 3rd instar larval wing disc of Drosophila are labeled by four fluorescent proteins using a Raeppli-NLS transgene, activated by tubulin-Gal4 and UAS-FLP. This approach facilitates the comprehensive analysis of clones in the entire tissue. (Reprinted with permission from Ref 10. Copyright 2014 The Company of Biologists Ltd.) Scale bar, 50 µm.
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fig05: Four examples of Brainbow technologies at work. (a) Purkinje cells in the mouse cerebellum are visualized in seven colors (i–vii) using Brainbow-3.1 and L7-Cre transgenes, as well as antibody amplification. (Reprinted with permission from Ref 16. Copyright 2013 Nature Publishing Group) Scale bar, 20 µm. (b) Pyramidal neurons in the P28 cortex of a CAG-CreERTM mouse are labeled by combinations of co-electroporated MAGIC Cytbow and Nucbow markers at E15. The image was acquired by two-photon microscopy. (Reprinted with permission from Ref 17. Copyright 2014 Elsevier Ltd.) Scale bar, 100 µm. (c) Neurites of lamina and medulla neuron subtypes (ln, mn) in the adult Drosophila optic lobe are visualized by endogenous fluorescent protein signals using a Flybow-2.0B transgene, activated by hs-mFLP5 and NP4151-Gal4—an enhancer trap insertion into the Netrin B locus. The image represents a single optical section. Several neurons (arrowheads) are suitable for tracing in stacks. Photoreceptor axons are visualized by immunolabeling with mAb24B10 (blue). Scale bar, 20 µm. (d) Nuclei of epithelial cell clones in a 3rd instar larval wing disc of Drosophila are labeled by four fluorescent proteins using a Raeppli-NLS transgene, activated by tubulin-Gal4 and UAS-FLP. This approach facilitates the comprehensive analysis of clones in the entire tissue. (Reprinted with permission from Ref 10. Copyright 2014 The Company of Biologists Ltd.) Scale bar, 50 µm.

Mentions: Brainbow methods were designed for anatomical and functional studies of genetically accessible cell populations with two main experimental applications in mind: (1) sparse labeling of specific cell types to visualize their morphologies and (2) comprehensive labeling of clonally related cells to track lineages (Figure 5). Consistently, Brainbow transgenes were so far successfully utilized to map known and new neuron subtypes,8,99 to identify the role of a basic helix-loop-helix transcription factor in axonal projection pattern formation,100 and to monitor laminar map assembly13 in the visual systems of flies and zebrafish. Furthermore, visualization of single cell shapes in their epithelial environment provided insights into the role of the tyrosine kinase Src42A in embryonic tracheal tube elongation.6 In lineage tracing experiments, Brainbow technologies were employed to follow the development of individual embryonic peripheral glial cell subtypes into perineurial, subperineurial, and wrapping glial subtypes associated with third instar larval peripheral nerves in Drosophila.101 Finally, multicolor clonal analysis discovered the contributions of dominant cardiomyocyte lineages to zebrafish heart morphogenesis,12 the role of neutral competition between symmetrically dividing intestinal crypt stem cells15 and the origin of stem cells required for corneal epithelial renewal in mice.102


Versatile genetic paintbrushes: Brainbow technologies.

Richier B, Salecker I - Wiley Interdiscip Rev Dev Biol (2014)

Four examples of Brainbow technologies at work. (a) Purkinje cells in the mouse cerebellum are visualized in seven colors (i–vii) using Brainbow-3.1 and L7-Cre transgenes, as well as antibody amplification. (Reprinted with permission from Ref 16. Copyright 2013 Nature Publishing Group) Scale bar, 20 µm. (b) Pyramidal neurons in the P28 cortex of a CAG-CreERTM mouse are labeled by combinations of co-electroporated MAGIC Cytbow and Nucbow markers at E15. The image was acquired by two-photon microscopy. (Reprinted with permission from Ref 17. Copyright 2014 Elsevier Ltd.) Scale bar, 100 µm. (c) Neurites of lamina and medulla neuron subtypes (ln, mn) in the adult Drosophila optic lobe are visualized by endogenous fluorescent protein signals using a Flybow-2.0B transgene, activated by hs-mFLP5 and NP4151-Gal4—an enhancer trap insertion into the Netrin B locus. The image represents a single optical section. Several neurons (arrowheads) are suitable for tracing in stacks. Photoreceptor axons are visualized by immunolabeling with mAb24B10 (blue). Scale bar, 20 µm. (d) Nuclei of epithelial cell clones in a 3rd instar larval wing disc of Drosophila are labeled by four fluorescent proteins using a Raeppli-NLS transgene, activated by tubulin-Gal4 and UAS-FLP. This approach facilitates the comprehensive analysis of clones in the entire tissue. (Reprinted with permission from Ref 10. Copyright 2014 The Company of Biologists Ltd.) Scale bar, 50 µm.
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Related In: Results  -  Collection

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fig05: Four examples of Brainbow technologies at work. (a) Purkinje cells in the mouse cerebellum are visualized in seven colors (i–vii) using Brainbow-3.1 and L7-Cre transgenes, as well as antibody amplification. (Reprinted with permission from Ref 16. Copyright 2013 Nature Publishing Group) Scale bar, 20 µm. (b) Pyramidal neurons in the P28 cortex of a CAG-CreERTM mouse are labeled by combinations of co-electroporated MAGIC Cytbow and Nucbow markers at E15. The image was acquired by two-photon microscopy. (Reprinted with permission from Ref 17. Copyright 2014 Elsevier Ltd.) Scale bar, 100 µm. (c) Neurites of lamina and medulla neuron subtypes (ln, mn) in the adult Drosophila optic lobe are visualized by endogenous fluorescent protein signals using a Flybow-2.0B transgene, activated by hs-mFLP5 and NP4151-Gal4—an enhancer trap insertion into the Netrin B locus. The image represents a single optical section. Several neurons (arrowheads) are suitable for tracing in stacks. Photoreceptor axons are visualized by immunolabeling with mAb24B10 (blue). Scale bar, 20 µm. (d) Nuclei of epithelial cell clones in a 3rd instar larval wing disc of Drosophila are labeled by four fluorescent proteins using a Raeppli-NLS transgene, activated by tubulin-Gal4 and UAS-FLP. This approach facilitates the comprehensive analysis of clones in the entire tissue. (Reprinted with permission from Ref 10. Copyright 2014 The Company of Biologists Ltd.) Scale bar, 50 µm.
Mentions: Brainbow methods were designed for anatomical and functional studies of genetically accessible cell populations with two main experimental applications in mind: (1) sparse labeling of specific cell types to visualize their morphologies and (2) comprehensive labeling of clonally related cells to track lineages (Figure 5). Consistently, Brainbow transgenes were so far successfully utilized to map known and new neuron subtypes,8,99 to identify the role of a basic helix-loop-helix transcription factor in axonal projection pattern formation,100 and to monitor laminar map assembly13 in the visual systems of flies and zebrafish. Furthermore, visualization of single cell shapes in their epithelial environment provided insights into the role of the tyrosine kinase Src42A in embryonic tracheal tube elongation.6 In lineage tracing experiments, Brainbow technologies were employed to follow the development of individual embryonic peripheral glial cell subtypes into perineurial, subperineurial, and wrapping glial subtypes associated with third instar larval peripheral nerves in Drosophila.101 Finally, multicolor clonal analysis discovered the contributions of dominant cardiomyocyte lineages to zebrafish heart morphogenesis,12 the role of neutral competition between symmetrically dividing intestinal crypt stem cells15 and the origin of stem cells required for corneal epithelial renewal in mice.102

Bottom Line: While being continuously refined, Brainbow technologies have thus found a firm place in the genetic toolboxes of developmental and neurobiologists.For further resources related to this article, please visit the WIREs website.The authors have declared no conflicts of interest for this article.

View Article: PubMed Central - PubMed

Affiliation: MRC National Institute for Medical Research, Division of Molecular Neurobiology, London, UK.

Show MeSH
Related in: MedlinePlus