Activation of Type 1 Cannabinoid Receptor (CB1R) Promotes Neurogenesis in Murine Subventricular Zone Cell Cultures
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{"title"=>"Activation of Type 1 Cannabinoid Receptor (CB1R) Promotes Neurogenesis in Murine Subventricular Zone Cell Cultures", "type"=>"journal", "authors"=>[{"first_name"=>"Sara", "last_name"=>"Xapelli", "scopus_author_id"=>"8721560200"}, {"first_name"=>"Fabienne", "last_name"=>"Agasse", "scopus_author_id"=>"23567488600"}, {"first_name"=>"Laura", "last_name"=>"Sardà-Arroyo", "scopus_author_id"=>"55185185800"}, {"first_name"=>"Liliana", "last_name"=>"Bernardino", "scopus_author_id"=>"23567485700"}, {"first_name"=>"Tiago", "last_name"=>"Santos", "scopus_author_id"=>"7004578673"}, {"first_name"=>"Filipa F.", "last_name"=>"Ribeiro", "scopus_author_id"=>"56818016300"}, {"first_name"=>"Jorge", "last_name"=>"Valero", "scopus_author_id"=>"8959759900"}, {"first_name"=>"José", "last_name"=>"Bragança", "scopus_author_id"=>"6602220001"}, {"first_name"=>"Clarissa", "last_name"=>"Schitine", "scopus_author_id"=>"23493561400"}, {"first_name"=>"Ricardo A.", "last_name"=>"de Melo Reis", "scopus_author_id"=>"35566644600"}, {"first_name"=>"Ana M.", "last_name"=>"Sebastião", "scopus_author_id"=>"7004409879"}, {"first_name"=>"João O.", "last_name"=>"Malva", "scopus_author_id"=>"35510123300"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"pui"=>"368960585", "sgr"=>"84877955877", "pmid"=>"23704915", "scopus"=>"2-s2.0-84877955877", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "doi"=>"10.1371/journal.pone.0063529", "issn"=>"19326203"}, "id"=>"bfaa3e7d-ba10-3191-8606-cc0090640927", "abstract"=>"The endocannabinoid system has been implicated in the modulation of adult neurogenesis. Here, we describe the effect of type 1 cannabinoid receptor (CB1R) activation on self-renewal, proliferation and neuronal differentiation in mouse neonatal subventricular zone (SVZ) stem/progenitor cell cultures. Expression of CB1R was detected in SVZ-derived immature cells (Nestin-positive), neurons and astrocytes. Stimulation of the CB1R by (R)-(+)-Methanandamide (R-m-AEA) increased self-renewal of SVZ cells, as assessed by counting the number of secondary neurospheres and the number of Sox2+/+ cell pairs, an effect blocked by Notch pathway inhibition. Moreover, R-m-AEA treatment for 48 h, increased proliferation as assessed by BrdU incorporation assay, an effect mediated by activation of MAPK-ERK and AKT pathways. Surprisingly, stimulation of CB1R by R-m-AEA also promoted neuronal differentiation (without affecting glial differentiation), at 7 days, as shown by counting the number of NeuN-positive neurons in the cultures. Moreover, by monitoring intracellular calcium concentrations ([Ca(2+)]i) in single cells following KCl and histamine stimuli, a method that allows the functional evaluation of neuronal differentiation, we observed an increase in neuronal-like cells. This proneurogenic effect was blocked when SVZ cells were co-incubated with R-m-AEA and the CB1R antagonist AM 251, for 7 days, thus indicating that this effect involves CB1R activation. In accordance with an effect on neuronal differentiation and maturation, R-m-AEA also increased neurite growth, as evaluated by quantifying and measuring the number of MAP2-positive processes. Taken together, these results demonstrate that CB1R activation induces proliferation, self-renewal and neuronal differentiation from mouse neonatal SVZ cell cultures.", "link"=>"http://www.mendeley.com/research/activation-type-1-cannabinoid-receptor-cb1r-promotes-neurogenesis-murine-subventricular-zone-cell-cu", "reader_count"=>38, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>2, "Librarian"=>1, "Researcher"=>4, "Student > Ph. D. Student"=>6, "Student > Master"=>10, "Other"=>4, "Student > Bachelor"=>9, "Professor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>2, "Librarian"=>1, "Researcher"=>4, "Student > Ph. D. Student"=>6, "Student > Master"=>10, "Other"=>4, "Student > Bachelor"=>9, "Professor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>4, "Engineering"=>2, "Biochemistry, Genetics and Molecular Biology"=>4, "Agricultural and Biological Sciences"=>15, "Medicine and Dentistry"=>1, "Neuroscience"=>5, "Arts and Humanities"=>1, "Pharmacology, Toxicology and Pharmaceutical Science"=>1, "Psychology"=>3, "Immunology and Microbiology"=>2}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>2}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>1}, "Neuroscience"=>{"Neuroscience"=>5}, "Psychology"=>{"Psychology"=>3}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>2}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>15}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>4}, "Unspecified"=>{"Unspecified"=>4}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}, "Arts and Humanities"=>{"Arts and Humanities"=>1}}, "reader_count_by_country"=>{"Portugal"=>1, "Germany"=>1, "Spain"=>2}, "group_count"=>3}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1065424"], "description"=>"<p><b>A:</b> Protocol used for studying cell proliferation. <b>B:</b> Bar graph depicts the numbers of BrdU-positive cells expressed as percentage of the total number of cells. Data are expressed as mean ± SEM. N = 3–5. **<i>P</i><0.01 using Dunnett’s multiple comparison test, for comparison with control. <b>C:</b> Representative digital images of BrdU (red nuclei)-positive cells and Hoechst 33342 staining (blue nuclei) in SVZ cultures. <b>D:</b> Amount of BrdU incorporation evaluated by densitometry (at 450 nm). N = 10. *P<0.05, **P<0.01 and ***<i>P</i><0.001 using Dunnnet’s multiple comparison test, for comparison with control; <sup>##</sup>P<0.01 and <sup>###</sup>P<0.001 using Dunnnet’s multiple comparison test, for comparison with R-m-AEA; Scale bar = 50 µm.</p>", "links"=>[], "tags"=>["developmental biology", "stem cells", "Neural stem cells", "Molecular cell biology", "Cellular types", "neuroscience", "Cellular neuroscience", "Neuronal morphology", "Developmental neuroscience", "Molecular neuroscience", "Signaling pathways", "Neurochemistry", "neuromodulation", "Neurobiology of disease and regeneration", "proliferation", "svz"], "article_id"=>705792, "categories"=>["Biological Sciences"], "users"=>["Sara Xapelli", "Fabienne Agasse", "Laura Sardà-Arroyo", "Liliana Bernardino", "Tiago Santos", "Filipa F. Ribeiro", "Jorge Valero", "Jose Braganca", "Clarissa Schitine", "Ricardo A. de Melo Reis", "Ana M. Sebastião", "João O. Malva"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0063529.g003", "stats"=>{"downloads"=>1, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_R_Methanandamide_induces_cell_proliferation_in_SVZ_cell_cultures_/705792", "title"=>"(R)-(+)-Methanandamide induces cell proliferation in SVZ cell cultures.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-05-21 01:36:32"}
  • {"files"=>["https://ndownloader.figshare.com/files/1065422"], "description"=>"<p><b>A:</b> Detection of CB1R by Western blotting in SVZ. Lane 1 corresponds to SVZ proliferating cells, lane 2 to SVZ extract from adult C57Bl6 mice and lane 3 to the negative control (total proteins from CB1R-KO mice). <b>B–F:</b> Representative confocal digital images depicting CB1R immunoreactivity in SVZ cells after 7 days of differentiation [CB1R (in red); nestin (in green), GFAP (in green), PSA-NCAM (in green), DCX (in green), βIII tubulin (in green), MAP2 (in green) and Hoechst 33342 (used to visualize cell nuclei, in blue)]. c1, e1 and f1 are magnifications of squares in C, E and F, respectively. Scale bars = 20 µm. SVZ: subventricular zone; GFAP: Glial fibrillary acidic protein; PSA-NCAM: Polysialylated neural cell adhesion; βIII tubulin: Neuron-specific class III beta-tubulin; MAP2: Microtubule-associated protein 2; CB1R: CB1 receptor.</p>", "links"=>[], "tags"=>["developmental biology", "stem cells", "Neural stem cells", "Molecular cell biology", "Cellular types", "neuroscience", "Cellular neuroscience", "Neuronal morphology", "Developmental neuroscience", "Molecular neuroscience", "Signaling pathways", "Neurochemistry", "neuromodulation", "Neurobiology of disease and regeneration", "cells"], "article_id"=>705790, "categories"=>["Biological Sciences"], "users"=>["Sara Xapelli", "Fabienne Agasse", "Laura Sardà-Arroyo", "Liliana Bernardino", "Tiago Santos", "Filipa F. Ribeiro", "Jorge Valero", "Jose Braganca", "Clarissa Schitine", "Ricardo A. de Melo Reis", "Ana M. Sebastião", "João O. Malva"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0063529.g001", "stats"=>{"downloads"=>0, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_SVZ_cells_express_CB1R_/705790", "title"=>"SVZ cells express CB1R.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-05-21 01:36:30"}
  • {"files"=>["https://ndownloader.figshare.com/files/1065433", "https://ndownloader.figshare.com/files/1065434", "https://ndownloader.figshare.com/files/1065435", "https://ndownloader.figshare.com/files/1065436", "https://ndownloader.figshare.com/files/1065437", "https://ndownloader.figshare.com/files/1065438"], "description"=>"<div><p>The endocannabinoid system has been implicated in the modulation of adult neurogenesis. Here, we describe the effect of type 1 cannabinoid receptor (CB1R) activation on self-renewal, proliferation and neuronal differentiation in mouse neonatal subventricular zone (SVZ) stem/progenitor cell cultures. Expression of CB1R was detected in SVZ-derived immature cells (Nestin-positive), neurons and astrocytes. Stimulation of the CB1R by (R)-(+)-Methanandamide (R-m-AEA) increased self-renewal of SVZ cells, as assessed by counting the number of secondary neurospheres and the number of Sox2+/+ cell pairs, an effect blocked by Notch pathway inhibition. Moreover, R-m-AEA treatment for 48 h, increased proliferation as assessed by BrdU incorporation assay, an effect mediated by activation of MAPK-ERK and AKT pathways. Surprisingly, stimulation of CB1R by R-m-AEA also promoted neuronal differentiation (without affecting glial differentiation), at 7 days, as shown by counting the number of NeuN-positive neurons in the cultures. Moreover, by monitoring intracellular calcium concentrations ([Ca<sup>2+</sup>]<sub>i</sub>) in single cells following KCl and histamine stimuli, a method that allows the functional evaluation of neuronal differentiation, we observed an increase in neuronal-like cells. This proneurogenic effect was blocked when SVZ cells were co-incubated with R-m-AEA and the CB1R antagonist AM 251, for 7 days, thus indicating that this effect involves CB1R activation. In accordance with an effect on neuronal differentiation and maturation, R-m-AEA also increased neurite growth, as evaluated by quantifying and measuring the number of MAP2-positive processes. Taken together, these results demonstrate that CB1R activation induces proliferation, self-renewal and neuronal differentiation from mouse neonatal SVZ cell cultures.</p></div>", "links"=>[], "tags"=>["developmental biology", "stem cells", "Neural stem cells", "Molecular cell biology", "Cellular types", "neuroscience", "Cellular neuroscience", "Neuronal morphology", "Developmental neuroscience", "Molecular neuroscience", "Signaling pathways", "Neurochemistry", "neuromodulation", "Neurobiology of disease and regeneration", "cannabinoid", "receptor", "neurogenesis", "murine", "subventricular"], "article_id"=>705801, "categories"=>["Biological Sciences"], "users"=>["Sara Xapelli", "Fabienne Agasse", "Laura Sardà-Arroyo", "Liliana Bernardino", "Tiago Santos", "Filipa F. Ribeiro", "Jorge Valero", "Jose Braganca", "Clarissa Schitine", "Ricardo A. de Melo Reis", "Ana M. Sebastião", "João O. Malva"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0063529.s001", "https://dx.doi.org/10.1371/journal.pone.0063529.s002", "https://dx.doi.org/10.1371/journal.pone.0063529.s003", "https://dx.doi.org/10.1371/journal.pone.0063529.s004", "https://dx.doi.org/10.1371/journal.pone.0063529.s005", "https://dx.doi.org/10.1371/journal.pone.0063529.s006"], "stats"=>{"downloads"=>1, "page_views"=>17, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Activation_of_Type_1_Cannabinoid_Receptor_CB1R_Promotes_Neurogenesis_in_Murine_Subventricular_Zone_Cell_Cultures_/705801", "title"=>"Activation of Type 1 Cannabinoid Receptor (CB1R) Promotes Neurogenesis in Murine Subventricular Zone Cell Cultures", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2013-05-21 01:36:41"}
  • {"files"=>["https://ndownloader.figshare.com/files/1065431"], "description"=>"<p>Primary antibodies used for immunocytochemistry.</p>", "links"=>[], "tags"=>["developmental biology", "stem cells", "Neural stem cells", "Molecular cell biology", "Cellular types", "neuroscience", "Cellular neuroscience", "Neuronal morphology", "Developmental neuroscience", "Molecular neuroscience", "Signaling pathways", "Neurochemistry", "neuromodulation", "Neurobiology of disease and regeneration", "antibodies"], "article_id"=>705799, "categories"=>["Biological Sciences"], "users"=>["Sara Xapelli", "Fabienne Agasse", "Laura Sardà-Arroyo", "Liliana Bernardino", "Tiago Santos", "Filipa F. Ribeiro", "Jorge Valero", "Jose Braganca", "Clarissa Schitine", "Ricardo A. de Melo Reis", "Ana M. Sebastião", "João O. Malva"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0063529.t001", "stats"=>{"downloads"=>1, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Primary_antibodies_used_for_immunocytochemistry_/705799", "title"=>"Primary antibodies used for immunocytochemistry.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-05-21 01:36:39"}
  • {"files"=>["https://ndownloader.figshare.com/files/1065430"], "description"=>"<p><b>A:</b> Schematic representation of the protocol. <b>B:</b> Bar graph depicts the numbers of either VGAT- or TH/βIII tubulin-positive cells, expressed as percentage of total cells. The data are expressed as percentage ± SEM. N = 3. *<i>P</i><0.05 using unpaired Student’s t test for comparison with control. <b>C:</b> Schematic representation of the protocol used for studying neuritogenesis. <b>D:</b> Representative confocal digital images of the GFP (green), MAP2 (red), Hoechst staining (blue), in control cultures and in cultures exposed to R-m-AEA. Scale bar = 20 µm. <b>E:</b> Bar graphs depict (from left to right): total length (µm), number of primary and number of secondary ramifications of MAP2 neurites per cell. N = 3. **P<0.01 using unpaired student’s t test for comparison with control. MAP2: Microtubule-associated protein 2; TH: tyrosine hydroxylase; βIII tubulin: Neuron-specific class III beta-tubulin; VGAT: vesicular GABA transporter.</p>", "links"=>[], "tags"=>["developmental biology", "stem cells", "Neural stem cells", "Molecular cell biology", "Cellular types", "neuroscience", "Cellular neuroscience", "Neuronal morphology", "Developmental neuroscience", "Molecular neuroscience", "Signaling pathways", "Neurochemistry", "neuromodulation", "Neurobiology of disease and regeneration", "differentiation", "gabaergic", "neurons"], "article_id"=>705798, "categories"=>["Biological Sciences"], "users"=>["Sara Xapelli", "Fabienne Agasse", "Laura Sardà-Arroyo", "Liliana Bernardino", "Tiago Santos", "Filipa F. Ribeiro", "Jorge Valero", "Jose Braganca", "Clarissa Schitine", "Ricardo A. de Melo Reis", "Ana M. Sebastião", "João O. Malva"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0063529.g008", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_R_Methanandamide_induces_the_differentiation_of_GABAergic_neurons_and_neuritogenesis_/705798", "title"=>"(R)-(+)-Methanandamide induces the differentiation of GABAergic neurons and neuritogenesis.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-05-21 01:36:38"}
  • {"files"=>["https://ndownloader.figshare.com/files/1065427"], "description"=>"<p><b>A:</b> Schematic representation of the protocol. <b>B:</b> Bar graph depicts the number of neuronal-like responding cells expressed as percentages of total cells analyzed by SCCI. N = 4–8. *P<0.05, ***P<0.01 using Dunnett’s multiple comparison test, for comparison with control. <b>C:</b> Representative SCCI profiles of response of about 20 cells in control, in R-m-AEA and in R-m-AEA+AM 251 treated cultures. <b>D</b>: Representative fluorescent digital images of NeuN-positive neurons (red) and Hoechst staining (blue nuclei). Scale bar = 50 µm. <b>E:</b> Bar graph depicts the number of NeuN-positive cells, expressed as the percentage of total cells <i>per</i> culture. Data are expressed as mean ± SEM. N = 5 ***P<0.001 using Dunnett’s multiple comparison test, for comparison with control. SCCI: single cell calcium imaging; NeuN: Neuronal Nuclei.</p>", "links"=>[], "tags"=>["developmental biology", "stem cells", "Neural stem cells", "Molecular cell biology", "Cellular types", "neuroscience", "Cellular neuroscience", "Neuronal morphology", "Developmental neuroscience", "Molecular neuroscience", "Signaling pathways", "Neurochemistry", "neuromodulation", "Neurobiology of disease and regeneration", "neuronal", "differentiation", "svz", "cultures", "cb1r"], "article_id"=>705795, "categories"=>["Biological Sciences"], "users"=>["Sara Xapelli", "Fabienne Agasse", "Laura Sardà-Arroyo", "Liliana Bernardino", "Tiago Santos", "Filipa F. Ribeiro", "Jorge Valero", "Jose Braganca", "Clarissa Schitine", "Ricardo A. de Melo Reis", "Ana M. Sebastião", "João O. Malva"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0063529.g005", "stats"=>{"downloads"=>1, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_R_Methanandamide_induces_neuronal_differentiation_in_SVZ_cultures_through_CB1R_activation_/705795", "title"=>"(R)-(+)-Methanandamide induces neuronal differentiation in SVZ cultures through CB1R activation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-05-21 01:36:35"}
  • {"files"=>["https://ndownloader.figshare.com/files/1065429"], "description"=>"<p><b>A:</b> Scheme of the protocol. <b>B:</b> Bar graph depicts the fold increase of H3K36m3 recruitment in the promoter region of <i>Ngn1</i> gene quantified by qChIP analysis. <b>C</b>: Bar graph depicts the fold increase of mRNA expression for Ngn1 protein evaluated by qRT-PCR analysis. Data are expressed as mean ± SEM. N = 4–7. *P<0.05, using Dunnett’s test for comparison with control (set to 1). H3K36m3: Histone H3 trimethylated on lysine 36; Ngn1: Neurogenin 1; qChIP: quantitative chromatin immunoprecipitation; qRT-PCR: quantitative real time polymerase chain reaction.</p>", "links"=>[], "tags"=>["developmental biology", "stem cells", "Neural stem cells", "Molecular cell biology", "Cellular types", "neuroscience", "Cellular neuroscience", "Neuronal morphology", "Developmental neuroscience", "Molecular neuroscience", "Signaling pathways", "Neurochemistry", "neuromodulation", "Neurobiology of disease and regeneration", "proneurogenic", "genes"], "article_id"=>705797, "categories"=>["Biological Sciences"], "users"=>["Sara Xapelli", "Fabienne Agasse", "Laura Sardà-Arroyo", "Liliana Bernardino", "Tiago Santos", "Filipa F. Ribeiro", "Jorge Valero", "Jose Braganca", "Clarissa Schitine", "Ricardo A. de Melo Reis", "Ana M. Sebastião", "João O. Malva"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0063529.g007", "stats"=>{"downloads"=>0, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_R_Methanandamide_promotes_the_expression_of_the_proneurogenic_genes_Ngn1_/705797", "title"=>"(R)-(+)-Methanandamide promotes the expression of the proneurogenic genes <i>Ngn1</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-05-21 01:36:37"}
  • {"files"=>["https://ndownloader.figshare.com/files/1065428"], "description"=>"<p><b>A, D:</b> Experimental protocol. <b>B, E:</b> Representative confocal digital images of BrdU (red), βIII tubulin (green) and Hoechst staining (blue) (B) and of BrdU (red), NeuN (green) and Hoechst staining (blue) (E). Scale bar = 20 µm. <b>C, F:</b> Bar graphs depict the number of βIII tubulin/BrdU-positive cells expressed as percentage of total βIII tubulin positive cells (C) and NeuN/BrdU-positive cells expressed as percentage of total NeuN-positive cells (F) per culture. Data are expressed as mean ± SEM. N = 3. *P<0.05, **P<0.01 using unpaired Student’s t test for comparison with control. βIII tubulin: Neuron-specific class III beta-tubulin; NeuN: Neuronal Nuclei.</p>", "links"=>[], "tags"=>["developmental biology", "stem cells", "Neural stem cells", "Molecular cell biology", "Cellular types", "neuroscience", "Cellular neuroscience", "Neuronal morphology", "Developmental neuroscience", "Molecular neuroscience", "Signaling pathways", "Neurochemistry", "neuromodulation", "Neurobiology of disease and regeneration", "proliferation"], "article_id"=>705796, "categories"=>["Biological Sciences"], "users"=>["Sara Xapelli", "Fabienne Agasse", "Laura Sardà-Arroyo", "Liliana Bernardino", "Tiago Santos", "Filipa F. Ribeiro", "Jorge Valero", "Jose Braganca", "Clarissa Schitine", "Ricardo A. de Melo Reis", "Ana M. Sebastião", "João O. Malva"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0063529.g006", "stats"=>{"downloads"=>0, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_R_Methanandamide_promotes_proliferation_of_neuroblasts_/705796", "title"=>"(R)-(+)-Methanandamide promotes proliferation of neuroblasts.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-05-21 01:36:36"}
  • {"files"=>["https://ndownloader.figshare.com/files/1065425"], "description"=>"<p><b>A:</b> Protocol used for studying glial differentiation. <b>B:</b> Western blot analysis of GFAP and Olig2 protein levels in SVZ. Data are expressed as mean ± SEM. N = 4. <b>C:</b> Bar graph depicts the number of GFAP and Olig2-positive cells, expressed as the percentage of total cells <i>per</i> culture. Data are expressed as mean ± SEM. N = 3. <b>D:</b> Representative fluorescent digital images of GFAP-positive cells (green), Olig2-positive cells (red) and Hoechst staining (blue nuclei). Scale bar = 50 µm.</p>", "links"=>[], "tags"=>["developmental biology", "stem cells", "Neural stem cells", "Molecular cell biology", "Cellular types", "neuroscience", "Cellular neuroscience", "Neuronal morphology", "Developmental neuroscience", "Molecular neuroscience", "Signaling pathways", "Neurochemistry", "neuromodulation", "Neurobiology of disease and regeneration", "induce", "glial", "differentiation", "svz", "cultures", "cb1r"], "article_id"=>705793, "categories"=>["Biological Sciences"], "users"=>["Sara Xapelli", "Fabienne Agasse", "Laura Sardà-Arroyo", "Liliana Bernardino", "Tiago Santos", "Filipa F. Ribeiro", "Jorge Valero", "Jose Braganca", "Clarissa Schitine", "Ricardo A. de Melo Reis", "Ana M. Sebastião", "João O. Malva"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0063529.g004", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_R_Methanandamide_does_not_induce_glial_differentiation_in_SVZ_cultures_through_CB1R_activation_/705793", "title"=>"(R)-(+)-Methanandamide does not induce glial differentiation in SVZ cultures through CB1R activation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-05-21 01:36:33"}
  • {"files"=>["https://ndownloader.figshare.com/files/1065423"], "description"=>"<p><b>A:</b> Experimental protocol. <b>B:</b> Bar graphs represent the number of primary and secondary neurospheres. Data are expressed as mean ± SEM. N = 6. *P<0.05, **P<0.01 and ***<i>P</i><0.001 using Dunnett’s multiple comparison test, for comparison with control; <sup>###</sup>P<0.001 using Dunnett’s multiple comparison test, for comparison with R-m-AEA. <b>C:</b> Protocol used for studying cell-fate. <b>D:</b> Confocal digital images of cell pairs obtained following (a) the symmetrical division of a SVZ cell into two Sox2+ cells (Sox2+/+), (b) the asymmetrical division into a Sox2+ and a Sox2- progenitor (Sox2+/−) and (c) the symmetrical terminal division into two Sox2- progenitors (Sox2−/−). Scale bars 20 µm. <b>E:</b> Bar graph illustrates the number of each type of cell divisions counted. Data are expressed as the percentage of total cell pairs and are represented as the mean ± SEM. N = 5. *P<0.05 and ***P<0.001 using Bonferroni’s multiple comparison test, for comparison with the respective controls; <sup>###</sup>P<0.001 using Bonferroni’s multiple comparison test, for comparison with the respective R-m-AEA. SOX2: sex determining region Y-box 2.</p>", "links"=>[], "tags"=>["developmental biology", "stem cells", "Neural stem cells", "Molecular cell biology", "Cellular types", "neuroscience", "Cellular neuroscience", "Neuronal morphology", "Developmental neuroscience", "Molecular neuroscience", "Signaling pathways", "Neurochemistry", "neuromodulation", "Neurobiology of disease and regeneration"], "article_id"=>705791, "categories"=>["Biological Sciences"], "users"=>["Sara Xapelli", "Fabienne Agasse", "Laura Sardà-Arroyo", "Liliana Bernardino", "Tiago Santos", "Filipa F. Ribeiro", "Jorge Valero", "Jose Braganca", "Clarissa Schitine", "Ricardo A. de Melo Reis", "Ana M. Sebastião", "João O. Malva"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0063529.g002", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_R_Methanandamide_promotes_self_renewal_/705791", "title"=>"(R)-(+)-Methanandamide promotes self-renewal.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-05-21 01:36:31"}

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Relative Metric

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