Essential Role of Neuron-Enriched Diacylglycerol Kinase (DGK), DGKβ in Neurite Spine Formation, Contributing to Cognitive Function
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{"title"=>"Essential role of neuron-enriched diacylglycerol kinase (DGK), DGKβ in neurite spine formation, contributing to cognitive function", "type"=>"journal", "authors"=>[{"first_name"=>"Yasuhito", "last_name"=>"Shirai", "scopus_author_id"=>"7202788791"}, {"first_name"=>"Takeshi", "last_name"=>"Kouzuki", "scopus_author_id"=>"36608307100"}, {"first_name"=>"Kenichi", "last_name"=>"Kakefuda", "scopus_author_id"=>"35077751400"}, {"first_name"=>"Shigeki", "last_name"=>"Moriguchi", "scopus_author_id"=>"9635375500"}, {"first_name"=>"Atsushi", "last_name"=>"Oyagi", "scopus_author_id"=>"23989394000"}, {"first_name"=>"Kyoji", "last_name"=>"Horie", "scopus_author_id"=>"7201734033"}, {"first_name"=>"Shin Ya", "last_name"=>"Morita", "scopus_author_id"=>"57200072774"}, {"first_name"=>"Masamitsu", "last_name"=>"Shimazawa", "scopus_author_id"=>"6602683783"}, {"first_name"=>"Kohji", "last_name"=>"Fukunaga", "scopus_author_id"=>"7201972807"}, {"first_name"=>"Junji", "last_name"=>"Takeda", "scopus_author_id"=>"7202209981"}, {"first_name"=>"Naoaki", "last_name"=>"Saito", "scopus_author_id"=>"7402349371"}, {"first_name"=>"Hideaki", "last_name"=>"Hara", "scopus_author_id"=>"56799750000"}], "year"=>2010, "source"=>"PLoS ONE", "identifiers"=>{"scopus"=>"2-s2.0-77955348076", "pmid"=>"20657643", "sgr"=>"77955348076", "doi"=>"10.1371/journal.pone.0011602", "issn"=>"19326203", "pui"=>"359319245"}, "id"=>"e71b3985-a953-3841-b794-e3fa2398ceee", "abstract"=>"BACKGROUND: Diacylglycerol (DG) kinase (DGK) phosphorylates DG to produce phosphatidic acid (PA). Of the 10 subtypes of mammalian DGKs, DGKbeta is a membrane-localized subtype and abundantly expressed in the cerebral cortex, hippocampus, and caudate-putamen. However, its physiological roles in neurons and higher brain function have not been elucidated.\\n\\nMETHODOLOGY/PRINCIPAL FINDINGS: We, therefore, developed DGKbeta KO mice using the Sleeping Beauty transposon system, and found that its long-term potentiation in the hippocampal CA1 region was reduced, causing impairment of cognitive functions including spatial and long-term memories in Y-maze and Morris water-maze tests. The primary cultured hippocampal neurons from KO mice had less branches and spines compared to the wild type. This morphological impairment was rescued by overexpression of DGKbeta. In addition, overexpression of DGKbeta in SH-SY5Y cells or primary cultured mouse hippocampal neurons resulted in branch- and spine-formation, while a splice variant form of DGKbeta, which has kinase activity but loses membrane localization, did not induce branches and spines. In the cells overexpressing DGKbeta but not the splice variant form, DGK product, PA, was increased and the substrate, DG, was decreased on the plasma membrane. Importantly, lower spine density and abnormality of PA and DG contents in the CA1 region of the KO mice were confirmed.\\n\\nCONCLUSIONS/SIGNIFICANCE: These results demonstrate that membrane-localized DGKbeta regulates spine formation by regulation of lipids, contributing to the maintenance of neural networks in synaptic transmission of cognitive processes including memory.", "link"=>"http://www.mendeley.com/research/essential-role-neuronenriched-diacylglycerol-kinase-dgk-dgk%CE%B2-neurite-spine-formation-contributing-co", "reader_count"=>24, "reader_count_by_academic_status"=>{"Librarian"=>1, "Researcher"=>3, "Student > Ph. D. Student"=>7, "Student > Postgraduate"=>2, "Student > Master"=>7, "Other"=>1, "Student > Bachelor"=>2, "Unspecified"=>1}, "reader_count_by_user_role"=>{"Librarian"=>1, "Researcher"=>3, "Student > Ph. D. Student"=>7, "Student > Postgraduate"=>2, "Student > Master"=>7, "Other"=>1, "Student > Bachelor"=>2, "Unspecified"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>2, "Biochemistry, Genetics and Molecular Biology"=>4, "Medicine and Dentistry"=>3, "Agricultural and Biological Sciences"=>9, "Neuroscience"=>3, "Business, Management and Accounting"=>1, "Psychology"=>1, "Chemistry"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>3}, "Neuroscience"=>{"Neuroscience"=>3}, "Chemistry"=>{"Chemistry"=>1}, "Psychology"=>{"Psychology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>9}, "Business, Management and Accounting"=>{"Business, Management and Accounting"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>4}, "Unspecified"=>{"Unspecified"=>2}}, "reader_count_by_country"=>{"Germany"=>1}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/839758"], "description"=>"<p>(A, B) Vector DNA at the donor site and Dgkβ locus. SA, splice acceptor; pA, polyadenylation signal; P, cytomegalovirus enhancer/chicken beta-actin chimeric promoter; SD, splice donor; gray arrows, inverted repeats/direct repeats for transposase-specific binding; B, Bgl II; N, Nco I; S, Spe I. (C, D) Southern blot analysis of gene from WT (+/+), heterozygous (Tp/+), and KO mouse (Tp/Tp). Genomic DNA was digested with BglII and NcoI and detected with the lacZ probe shown in black. A single band in mutant mice indicates a single transposon insertion site segregated from the donor site (C). Similarly, genome DNA digested with Spe I was detected with the probe shown in red. Control represents genome from a control mouse cell line with one copy of neo. No band in the mutant mouse confirmed segregation of the donor site (D). (E) Typical result of PCR for genotyping. Bands at 455 bp and 540 bp are expected from the mutant and WT alleles, respectively. (F) RT-PCR. No band in RT-PCR indicates no mRNA of DGKβ in KO mice. (G) Western blotting using anti-DGKβ antibody. Proteins in the homogenate of hind- or fore-brain from WT and KO mice were separated by 7.5% SDS-PAGE, followed by transferring and immunostaining. (H) Immunohistochemistry using DGKβ antibody and frontal sections from WT and KO mice. (I) X-gal staining. The regions where DGKβ gene was mutated were determined by incubation with X-gal. Representative images showing cortex (ctx), caudate putamen (CP), and hippocampus (hip) of WT mice (upper) and DGKβKO mice (lower). Scale bar = 500 µm.</p>", "links"=>[], "tags"=>["ko"], "article_id"=>510208, "categories"=>["Neuroscience"], "users"=>["Yasuhito Shirai", "Takeshi Kouzuki", "Kenichi Kakefuda", "Shigeki Moriguchi", "Atsushi Oyagi", "Kyoji Horie", "Shin-ya Morita", "Masamitsu Shimazawa", "Kohji Fukunaga", "Junji Takeda", "Naoaki Saito", "Hideaki Hara"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0011602.g001", "stats"=>{"downloads"=>2, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Characterization_of_DGK_946_KO_mice_/510208", "title"=>"Characterization of DGKβ KO mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-07-15 00:03:28"}
  • {"files"=>["https://ndownloader.figshare.com/files/840116"], "description"=>"<p>Hippocampal neurons from WT and KO mice were cultured for indicated days. After fixing, the neurons were immunostained with MAP-2 followed by Alexa488-conjugated secondary antibody, and observed under confocal microscopy. (A) Typical images. Upper panels show lower magnification images and lower panels are magnified ones. Bars are 20 µm. (B) Static analysis. Following numbers of samples were analyzed: day 3, WT, n = 28 and KO, n = 27; day 10, WT, n = 37 and KO, n = 49; day 15, WT, n = 34 and KO, n = 31. *, **, and *** represent P<0.05, P<0.01, and P<0.005 vs. the control of WT, respectively.</p>", "links"=>[], "tags"=>["neuronal", "branching", "cultured", "hippocampal", "neurons", "ko"], "article_id"=>510558, "categories"=>["Neuroscience"], "users"=>["Yasuhito Shirai", "Takeshi Kouzuki", "Kenichi Kakefuda", "Shigeki Moriguchi", "Atsushi Oyagi", "Kyoji Horie", "Shin-ya Morita", "Masamitsu Shimazawa", "Kohji Fukunaga", "Junji Takeda", "Naoaki Saito", "Hideaki Hara"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0011602.g004", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Impairment_of_neuronal_branching_in_the_primary_cultured_hippocampal_neurons_from_DGK_946_KO_mice_/510558", "title"=>"Impairment of neuronal branching in the primary cultured hippocampal neurons from DGKβ KO mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-07-15 00:09:18"}
  • {"files"=>["https://ndownloader.figshare.com/files/839965"], "description"=>"<p>(A) Representative fEPSPs recorded from the CA1 region. (B) Changes in slopes of fEPSPs following HFS in the CA1 region from wild-type and DGKβ KO mice. (C) Level of LTP potentiation 60 min after HFS in the CA1 region from WT and DGKβ-KO mice. ** represents P<0.01 vs. the control of WT mice. (D) Input-output relationship between WT and KO mice. In the input-output relationship, amplitude of fEPSPs by stimulus intensity at 0.1 mA to 1.0 mA did not affect between DGKβ-KO mice and wild-type mice.</p>", "links"=>[], "tags"=>["potentiation", "ko"], "article_id"=>510409, "categories"=>["Neuroscience"], "users"=>["Yasuhito Shirai", "Takeshi Kouzuki", "Kenichi Kakefuda", "Shigeki Moriguchi", "Atsushi Oyagi", "Kyoji Horie", "Shin-ya Morita", "Masamitsu Shimazawa", "Kohji Fukunaga", "Junji Takeda", "Naoaki Saito", "Hideaki Hara"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0011602.g003", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Impaired_long_term_potentiation_LTP_in_the_DGK_946_KO_mice_/510409", "title"=>"Impaired long-term potentiation (LTP) in the DGKβ KO mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-07-15 00:06:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/839848"], "description"=>"<p>(A) Spontaneous alteration behavior of Y-maze test. WT (n = 9) and DGKβ KO (n = 7) mice were placed at the end of one fixed arm and allowed to move freely through the maze during an 8-min session. ** represents P<0.01 vs. WT in the Student's <i>t</i>-test. (B and C) Morris water maze test of DGKβ KO mice (n = 9) and their WT litter mates (n = 8). Mice were placed in the opposite quadrant facing the wall and the time spent in each quadrant (B) and the mean distance from the platform (C) were measured. * indicates that the period DGKβ KO mice spent in the target quadrant was significantly different from that of WT (P<0.05, Student's t-test). # represents the probability that the difference in the time spent in each quadrant versus the target quadrant was significant (P<0.05 with one-way ANOVA and followed by Tukey's post hoc test.)</p>", "links"=>[], "tags"=>["ko"], "article_id"=>510293, "categories"=>["Neuroscience"], "users"=>["Yasuhito Shirai", "Takeshi Kouzuki", "Kenichi Kakefuda", "Shigeki Moriguchi", "Atsushi Oyagi", "Kyoji Horie", "Shin-ya Morita", "Masamitsu Shimazawa", "Kohji Fukunaga", "Junji Takeda", "Naoaki Saito", "Hideaki Hara"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0011602.g002", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Impaired_cognitive_function_in_the_DGK_946_KO_mice_/510293", "title"=>"Impaired cognitive function in the DGKβ KO mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-07-15 00:04:53"}
  • {"files"=>["https://ndownloader.figshare.com/files/840672"], "description"=>"<p>(A) Typical Golgi staining of hippocampal neurons at CA1 regions. Yellow arrows show spines. Scale bar represents 5 µm. (B) Comparison of synapse density in the CA1 hippocampal region. The number of synapses was counted and plotted. Each datum point represents the mean and SEM (WT, n = 14; KO, n = 18). ** represents P<0.01 vs. the control of WT. (C) Typical images of electron microscopy. (×243,000) Scale bar represents 5 µm. (D) Comparison of the number of synaptic junctions. Number of synaptic junctions with PSD in the micrographs was counted. Each datum point represents the mean and SEM (WT, n = 40; KO, n = 40). Red arrows indicate synaptic junctions. *** represents P<0.005 vs. the control of WT. (E, F) PA or DG level in the hippocampus and cerebellum from WT or KO mice. The hippocampus and cerebellum were dissected from WT or KO mice, and PA and DG contents were measured as described in the <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0011602#s4\" target=\"_blank\">Materials and Methods</a>. Each datum point represents mean with STD of three independent experiments. * and *** mean P<0.05 and P<0.005 vs. WT (Student's T-test).</p>", "links"=>[], "tags"=>["synapse", "levels", "hippocampus", "ko"], "article_id"=>511107, "categories"=>["Neuroscience"], "users"=>["Yasuhito Shirai", "Takeshi Kouzuki", "Kenichi Kakefuda", "Shigeki Moriguchi", "Atsushi Oyagi", "Kyoji Horie", "Shin-ya Morita", "Masamitsu Shimazawa", "Kohji Fukunaga", "Junji Takeda", "Naoaki Saito", "Hideaki Hara"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0011602.g009", "stats"=>{"downloads"=>1, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Abnormality_of_synapse_density_and_PA_DG_levels_in_the_hippocampus_of_DGK_946_KO_mice_/511107", "title"=>"Abnormality of synapse density and PA/DG levels in the hippocampus of DGKβ KO mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-07-15 00:18:27"}
  • {"files"=>["https://ndownloader.figshare.com/files/418884", "https://ndownloader.figshare.com/files/418914", "https://ndownloader.figshare.com/files/418956", "https://ndownloader.figshare.com/files/418986"], "description"=>"<div><h3>Background</h3><p>Diacylglycerol (DG) kinase (DGK) phosphorylates DG to produce phosphatidic acid (PA). Of the 10 subtypes of mammalian DGKs, DGKβ is a membrane-localized subtype and abundantly expressed in the cerebral cortex, hippocampus, and caudate-putamen. However, its physiological roles in neurons and higher brain function have not been elucidated.</p> <h3>Methodology/Principal Findings</h3><p>We, therefore, developed DGKβ KO mice using the Sleeping Beauty transposon system, and found that its long-term potentiation in the hippocampal CA1 region was reduced, causing impairment of cognitive functions including spatial and long-term memories in Y-maze and Morris water-maze tests. The primary cultured hippocampal neurons from KO mice had less branches and spines compared to the wild type. This morphological impairment was rescued by overexpression of DGKβ. In addition, overexpression of DGKβ in SH-SY5Y cells or primary cultured mouse hippocampal neurons resulted in branch- and spine-formation, while a splice variant form of DGKβ, which has kinase activity but loses membrane localization, did not induce branches and spines. In the cells overexpressing DGKβ but not the splice variant form, DGK product, PA, was increased and the substrate, DG, was decreased on the plasma membrane. Importantly, lower spine density and abnormality of PA and DG contents in the CA1 region of the KO mice were confirmed.</p> <h3>Conclusions/Significance</h3><p>These results demonstrate that membrane-localized DGKβ regulates spine formation by regulation of lipids, contributing to the maintenance of neural networks in synaptic transmission of cognitive processes including memory.</p> </div>", "links"=>[], "tags"=>["neuron-enriched", "diacylglycerol", "kinase", "neurite", "contributing"], "article_id"=>142722, "categories"=>["Neuroscience"], "users"=>["Yasuhito Shirai", "Takeshi Kouzuki", "Kenichi Kakefuda", "Shigeki Moriguchi", "Atsushi Oyagi", "Kyoji Horie", "Shin-ya Morita", "Masamitsu Shimazawa", "Kohji Fukunaga", "Junji Takeda", "Naoaki Saito", "Hideaki Hara"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0011602.s001", "https://dx.doi.org/10.1371/journal.pone.0011602.s002", "https://dx.doi.org/10.1371/journal.pone.0011602.s003", "https://dx.doi.org/10.1371/journal.pone.0011602.s004"], "stats"=>{"downloads"=>4, "page_views"=>19, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Essential_Role_of_Neuron_Enriched_Diacylglycerol_Kinase_DGK_DGK_in_Neurite_Spine_Formation_Contributing_to_Cognitive_Function/142722", "title"=>"Essential Role of Neuron-Enriched Diacylglycerol Kinase (DGK), DGKβ in Neurite Spine Formation, Contributing to Cognitive Function", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2010-07-15 00:45:22"}
  • {"files"=>["https://ndownloader.figshare.com/files/840531"], "description"=>"<p>(A) Typical images of SH-SY5Y cells overexpressing GFP-DGKβ, GFP-C-cut or GFP alone. Upper panels show lower magnification images and lower panels are magnified ones. Bars are 20 µm. Arrowheads indicate the position of spine-like structures. (B) Statistical analysis of morphological differences between SH-SY5Y cells overexpressing GFP-DGKβ, GFP-C-cut or GFP alone. More than 100 cells were observed in each experiment and three independent experiments were performed. The mean and SEM of number of the cells with one or two long neurites (closed column), several neurites with branches (slashed column), and no neurites (open column) are shown as percentage to the transfected cells. (C, D) Comparison of PA and DG level on the membrane of SH-SY5Y cells expressing GFP-DGKβ, GFP-C-cut or GFP alone. Membrane PA and DG were extracted from the cells and measured as described in the <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0011602#s4\" target=\"_blank\">Materials and Methods</a>. PA or DG contents are represented as mean of percentage to those of control cells expressing GFP alone. Each datum points represents mean with STD of three independent experiments. * means P<0.05 vs. the control expressing GFP alone (Student's T-test).</p>", "links"=>[], "tags"=>["splice", "variant", "induce", "branches", "spines", "membrane", "pa", "dg"], "article_id"=>510969, "categories"=>["Neuroscience"], "users"=>["Yasuhito Shirai", "Takeshi Kouzuki", "Kenichi Kakefuda", "Shigeki Moriguchi", "Atsushi Oyagi", "Kyoji Horie", "Shin-ya Morita", "Masamitsu Shimazawa", "Kohji Fukunaga", "Junji Takeda", "Naoaki Saito", "Hideaki Hara"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0011602.g008", "stats"=>{"downloads"=>1, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Disability_of_DGK_946_splice_variant_form_C_cut_to_induce_branches_and_spines_and_its_correlation_to_membrane_PA_and_DG_levels_/510969", "title"=>"Disability of DGKβ splice variant form (C-cut) to induce branches and spines and its correlation to membrane PA and DG levels.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-07-15 00:16:09"}
  • {"files"=>["https://ndownloader.figshare.com/files/840337"], "description"=>"<p>The mouse hippocampal neurons cultured for 3 days (A, B) or 10 days (C, D) were infected with respective viruses. After 48 h of infection, the cells were observed under confocal microscopy and analyzed using Neurolucida soft ware. (A) Typical images of primary cultured hippocampal neurons overexpressing GFP-DGKβ or GFP alone. (B) Statistical analysis of number of branches per a single neurite. n = 13 for GFP, n = 8 for DGKβ. (C) Typical images of spine-like structures in the primary cultured hippocampal neurons overexpressing GFP-DGKβ or GFP alone. Magnified images of squared area are shown in right panels. (D) Statistical analysis of spine-like structures. n = 18 for GFP and n = 15 for DGKβ. *** means P<0.005 vs. the control expressing GFP alone. Bars represent 5 µm (for magnified images) or 20 µm.</p>", "links"=>[], "tags"=>["branching", "cultured", "hippocampal", "neurons", "wt"], "article_id"=>510775, "categories"=>["Neuroscience"], "users"=>["Yasuhito Shirai", "Takeshi Kouzuki", "Kenichi Kakefuda", "Shigeki Moriguchi", "Atsushi Oyagi", "Kyoji Horie", "Shin-ya Morita", "Masamitsu Shimazawa", "Kohji Fukunaga", "Junji Takeda", "Naoaki Saito", "Hideaki Hara"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0011602.g006", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_DGK_946_induced_branching_and_spine_formation_in_the_primary_cultured_hippocampal_neurons_from_WT_mice_/510775", "title"=>"DGKβ-induced branching and spine formation in the primary cultured hippocampal neurons from WT mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-07-15 00:12:55"}
  • {"files"=>["https://ndownloader.figshare.com/files/840430"], "description"=>"<p>(A) Typical images of WT and KO primary cultured hippocampal neurons expressing GFP or GFP-DGKβ, and its distal dendrites with spines (upper panels). Bars are 20 µm or 5 µm (for upper images). (B, C) Comparison between numbers of branches per single neurite (B), and number of spines (C) in primary cultured hippocampal neurons from WT and KO mice. Numbers of analyzed samples are shown in the graph. **, and *** represent P<0.01 and P<0.005, respectively.</p>", "links"=>[], "tags"=>["impaired", "branching", "cultured", "hippocampal", "neurons", "ko", "mice"], "article_id"=>510870, "categories"=>["Neuroscience"], "users"=>["Yasuhito Shirai", "Takeshi Kouzuki", "Kenichi Kakefuda", "Shigeki Moriguchi", "Atsushi Oyagi", "Kyoji Horie", "Shin-ya Morita", "Masamitsu Shimazawa", "Kohji Fukunaga", "Junji Takeda", "Naoaki Saito", "Hideaki Hara"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0011602.g007", "stats"=>{"downloads"=>1, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Rescue_of_impaired_branching_and_spine_formation_in_the_primary_cultured_hippocampal_neurons_from_KO_mice_by_DGK_946_overexpression_/510870", "title"=>"Rescue of impaired branching and spine formation in the primary cultured hippocampal neurons from KO mice by DGKβ overexpression.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-07-15 00:14:30"}
  • {"files"=>["https://ndownloader.figshare.com/files/840231"], "description"=>"<p>Hippocampal neurons from WT and KO mice were cultured for 14 days. After fixing, the neurons were immunostained with PSD-95 (green) and MAP-2 (red) antibodies followed by Alexa488- and Alexa-594-conjugated secondary antibody. Following numbers of samples were analyzed: WT, n = 36 and KO, n = 50; *** represents P<0.005 vs. the control of WT, respectively.</p>", "links"=>[], "tags"=>["cultured", "hippocampal", "neurons", "ko"], "article_id"=>510675, "categories"=>["Neuroscience"], "users"=>["Yasuhito Shirai", "Takeshi Kouzuki", "Kenichi Kakefuda", "Shigeki Moriguchi", "Atsushi Oyagi", "Kyoji Horie", "Shin-ya Morita", "Masamitsu Shimazawa", "Kohji Fukunaga", "Junji Takeda", "Naoaki Saito", "Hideaki Hara"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0011602.g005", "stats"=>{"downloads"=>1, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Impairment_of_spine_formation_in_the_primary_cultured_hippocampal_neurons_from_DGK_946_KO_mice_/510675", "title"=>"Impairment of spine formation in the primary cultured hippocampal neurons from DGKβ KO mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-07-15 00:11:15"}

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  • {"unique-ip"=>"9", "full-text"=>"8", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"4", "cited-by"=>"0", "year"=>"2019", "month"=>"4"}
  • {"unique-ip"=>"6", "full-text"=>"7", "pdf"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"5"}

Relative Metric

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