Expression Analysis of CB2-GFP BAC Transgenic Mice
Publication Date
September 25, 2015
Journal
PLOS ONE
Authors
Anne Caroline Schmöle, Ramona Lundt, Benjamin Gennequin, Hanna Schrage, et al
Volume
10
Issue
9
Pages
e0138986
DOI
https://dx.plos.org/10.1371/journal.pone.0138986
Publisher URL
http://journals.plos.org/plosone/article?id=10.1371%2Fjournal.pone.0138986
PubMed
http://www.ncbi.nlm.nih.gov/pubmed/26406232
PubMed Central
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4583291
Europe PMC
http://europepmc.org/abstract/MED/26406232
Web of Science
000361800700138
Scopus
84947252004
Mendeley
http://www.mendeley.com/research/expression-analysis-cb2gfp-bac-transgenic-mice
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Mendeley | Further Information

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Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/2289145"], "description"=>"<p>Splenocytes of WT and CB2-GFPTg mice were stimulated with LPS or CpG and GFP expression was analyzed by flow cytometry. GFP expression is significantly enhanced after stimulation in CB2-GFP mice (a,c) but not in WT mice (a). The number of B220<sup>+</sup> cells is not altered by stimulation with LPS or CpG (b). N = 3, all samples were analyzed in triplicates. Data were analyzed by Students T-Test, *p < 0.05, **p < 0.001, to unstimulated genotype control.</p>", "links"=>[], "tags"=>["CB 2 expression", "CB 2 antibodies", "display GFP expression", "CB 2 promoter", "CB 2 involvement", "Cannabinoid receptor 1", "b cells", "ecs", "reporter mouse line", "bac", "GFP reporter mice", "trace CB 2 expression", "CB 2", "study CB 2 expression"], "article_id"=>1556769, "categories"=>["Biological Sciences"], "users"=>["Anne-Caroline Schmöle", "Ramona Lundt", "Benjamin Gennequin", "Hanna Schrage", "Eva Beins", "Alexandra Krämer", "Till Zimmer", "Andreas Limmer", "Andreas Zimmer", "David-Marian Otte"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0138986.g005", "stats"=>{"downloads"=>0, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Enhanced_GFP_expression_in_stimulated_splenic_B_cells_/1556769", "title"=>"Enhanced GFP expression in stimulated splenic B cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-09-25 03:34:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/2289147", "https://ndownloader.figshare.com/files/2289148", "https://ndownloader.figshare.com/files/2289149", "https://ndownloader.figshare.com/files/2289150"], "description"=>"<div><p>The endocannabinoid system (ECS) is a retrograde messenger system, consisting of lipid signaling molecules that bind to at least two G-protein-coupled receptors, Cannabinoid receptor 1 and 2 (CB1 and 2). As CB2 is primarily expressed on immune cells such as B cells, T cells, macrophages, dendritic cells, and microglia, it is of great interest how CB2 contributes to immune cell development and function in health and disease. Here, understanding the mechanisms of CB2 involvement in immune-cell function as well as the trafficking and regulation of CB2 expressing cells are crucial issues. Up to now, CB2 antibodies produce unclear results, especially those targeting the murine protein. Therefore, we have generated BAC transgenic GFP reporter mice (CB2-GFPTg) to trace CB2 expression <i>in vitro</i> and <i>in situ</i>. Those mice express GFP under the CB2 promoter and display GFP expression paralleling CB2 expression on the transcript level in spleen, thymus and brain tissue. Furthermore, by using fluorescence techniques we show that the major sources for GFP-CB2 expression are B cells in spleen and blood and microglia in the brain. This novel CB2-GFP transgenic reporter mouse line represents a powerful resource to study CB2 expression in different cell types. Furthermore, it could be used for analyzing CB2-mediated mobilization and trafficking of immune cells as well as studying the fate of recruited immune cells in models of acute and chronic inflammation.</p></div>", "links"=>[], "tags"=>["CB 2 expression", "CB 2 antibodies", "display GFP expression", "CB 2 promoter", "CB 2 involvement", "Cannabinoid receptor 1", "b cells", "ecs", "reporter mouse line", "bac", "GFP reporter mice", "trace CB 2 expression", "CB 2", "study CB 2 expression"], "article_id"=>1556771, "categories"=>["Biological Sciences"], "users"=>["Anne-Caroline Schmöle", "Ramona Lundt", "Benjamin Gennequin", "Hanna Schrage", "Eva Beins", "Alexandra Krämer", "Till Zimmer", "Andreas Limmer", "Andreas Zimmer", "David-Marian Otte"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0138986.s001", "https://dx.doi.org/10.1371/journal.pone.0138986.s002", "https://dx.doi.org/10.1371/journal.pone.0138986.s003", "https://dx.doi.org/10.1371/journal.pone.0138986.s004"], "stats"=>{"downloads"=>4, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Expression_Analysis_of_CB2_GFP_BAC_Transgenic_Mice_/1556771", "title"=>"Expression Analysis of CB2-GFP BAC Transgenic Mice", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2015-09-25 03:34:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/2289132"], "description"=>"<p>(a) Schematic diagram of GFP-FRT-Neo® BAC modification fragment and homologous recombination into the wild-type Cnr2 BAC depicts Cnr2 exons (black rectangles), ORF (open box), and GFP-FRT-Neo® fusion reporter (green/blue). Cnr2 homology arms (each ∼50 bp) are shown on either side of the insert sequence. Introduction of the GFP-FRT-Neo® cassette leads to a replacement of the Cnr2 ORF without affecting any putative Cnr2 promoter sequences. Excision of the Neo® cassette after integration to derive the final CB2-GFPTg BAC transgene is shown. Restriction enzymes and a genomic probe (black bar) used for <i>Southern blot</i> analysis are indicated. Exons are represented as rectangles and FRT sites as triangles. P: <i>Pst</i>I; A: <i>Ase</i>I; C: <i>Cla</i>I. (b) Screening for transgene integration by PCR. Integration of the CB2-GFPTg BAC into the genomic mouse DNA was confirmed by PCR with specific primers (eGFP_Aat_F2 and eGFP_R2) as indicated with arrows in a. Three independent founders were identified (asterisks) by the detection of an additional band representing the GFP. (c) Comparison of GFP and CB2 mRNA expression in brain, spleen and thymus relative to HPRT reference gene expression. In brain tissue samples neither GFP nor CB2 mRNA expression was detectable. Thymus showed moderate expression of both, GFP and CB2, whereas spleen tissue revealed highest expression of GFP and CB2. n = 4. (d) Representative western blot analysis of GFP protein expression in brain, thymus and spleen in CB2-GFPTg and WT mice. CB2-GFPtg mice showed moderate GFP protein expression in thymus and high GFP protein expression in spleen. No detectable GFP protein expression in brain tissue samples as well as in samples of WT littermates. GAPDH was used as loading control.</p>", "links"=>[], "tags"=>["CB 2 expression", "CB 2 antibodies", "display GFP expression", "CB 2 promoter", "CB 2 involvement", "Cannabinoid receptor 1", "b cells", "ecs", "reporter mouse line", "bac", "GFP reporter mice", "trace CB 2 expression", "CB 2", "study CB 2 expression"], "article_id"=>1556757, "categories"=>["Biological Sciences"], "users"=>["Anne-Caroline Schmöle", "Ramona Lundt", "Benjamin Gennequin", "Hanna Schrage", "Eva Beins", "Alexandra Krämer", "Till Zimmer", "Andreas Limmer", "Andreas Zimmer", "David-Marian Otte"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0138986.g001", "stats"=>{"downloads"=>4, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Construction_of_CB2_GFP_BAC_transgene_and_expression_in_various_transgenic_mouse_tissues_/1556757", "title"=>"Construction of CB2-GFP BAC transgene and expression in various transgenic mouse tissues.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-09-25 03:34:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/2289134"], "description"=>"<p>Expression of CB2 protein was investigated using CB2-GFP mice and depicted by representative histograms of three individual transgenic mice in comparison to a non-transgenic littermate. In average 23% of CD4<sup>+</sup> T-lymphocytes (a), 53% of CD8 T-lymphocytes (b), 64% of CD11b (c), 46% of CD19 (d) and 30% of CD335 cells (e) showed expression of GFP. All subsets were pre-gated on CD45+ cells and data analysis was performed using FlowJo software (Tree Star Inc., Ashland, USA).</p>", "links"=>[], "tags"=>["CB 2 expression", "CB 2 antibodies", "display GFP expression", "CB 2 promoter", "CB 2 involvement", "Cannabinoid receptor 1", "b cells", "ecs", "reporter mouse line", "bac", "GFP reporter mice", "trace CB 2 expression", "CB 2", "study CB 2 expression"], "article_id"=>1556759, "categories"=>["Biological Sciences"], "users"=>["Anne-Caroline Schmöle", "Ramona Lundt", "Benjamin Gennequin", "Hanna Schrage", "Eva Beins", "Alexandra Krämer", "Till Zimmer", "Andreas Limmer", "Andreas Zimmer", "David-Marian Otte"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0138986.g002", "stats"=>{"downloads"=>2, "page_views"=>24, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Comparison_of_GFP_expression_in_murine_PBMC_using_flow_cytometry_/1556759", "title"=>"Comparison of GFP expression in murine PBMC using flow cytometry.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-09-25 03:34:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/2289138"], "description"=>"<p>Immunohistochemical analysis of GFP expression in the spleen of CB2-GFPTg mice. Shown are representative spleen tissue sections of CB2-GFPTg stained for B220, CD11c, Iba1 and TCRβ (red), GFP expression is displayed as green staining. CB2-GFP is co-localized with B220 expression, whereas no co-expression with CD11c, Iba1 or TCRβ was observed (scale bar = 50 μm).</p>", "links"=>[], "tags"=>["CB 2 expression", "CB 2 antibodies", "display GFP expression", "CB 2 promoter", "CB 2 involvement", "Cannabinoid receptor 1", "b cells", "ecs", "reporter mouse line", "bac", "GFP reporter mice", "trace CB 2 expression", "CB 2", "study CB 2 expression"], "article_id"=>1556763, "categories"=>["Biological Sciences"], "users"=>["Anne-Caroline Schmöle", "Ramona Lundt", "Benjamin Gennequin", "Hanna Schrage", "Eva Beins", "Alexandra Krämer", "Till Zimmer", "Andreas Limmer", "Andreas Zimmer", "David-Marian Otte"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0138986.g003", "stats"=>{"downloads"=>5, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Analysis_of_CB2_expression_in_CB2_GFPTg_mice_/1556763", "title"=>"Analysis of CB2 expression in CB2-GFPTg mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-09-25 03:34:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/2289142"], "description"=>"<p>Immunohistochemical analysis of GFP expression in the brain of CB2-GFPTg mice. CB2-GFPtg mice show GFP expression in the hippocampus (a, right) which is not present in WT mice (a, left) (scale bar = 250 μm). CB2-GFP expression is co-localized with Iba1-staining (b, second row), but not with NeuN (b, third row) or GFAP (b, fourth row). <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0138986#pone.0138986.g004\" target=\"_blank\">Fig 4B</a>, first row shows background analysis in WT mice (scale bar = 50 μm).</p>", "links"=>[], "tags"=>["CB 2 expression", "CB 2 antibodies", "display GFP expression", "CB 2 promoter", "CB 2 involvement", "Cannabinoid receptor 1", "b cells", "ecs", "reporter mouse line", "bac", "GFP reporter mice", "trace CB 2 expression", "CB 2", "study CB 2 expression"], "article_id"=>1556767, "categories"=>["Biological Sciences"], "users"=>["Anne-Caroline Schmöle", "Ramona Lundt", "Benjamin Gennequin", "Hanna Schrage", "Eva Beins", "Alexandra Krämer", "Till Zimmer", "Andreas Limmer", "Andreas Zimmer", "David-Marian Otte"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0138986.g004", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_CB2_GFP_expression_in_the_hippocampus_of_WT_and_CB2_GFP_mice_/1556767", "title"=>"CB2-GFP expression in the hippocampus of WT and CB2-GFP mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-09-25 03:34:07"}

PMC Usage Stats | Further Information

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{"start_date"=>"2015-01-01T00:00:00Z", "end_date"=>"2015-12-31T00:00:00Z", "subject_areas"=>[]}
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