Loss of the SV2-like Protein SVOP Produces No Apparent Deficits in Laboratory Mice
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{"title"=>"Loss of the SV2-like Protein SVOP Produces No Apparent Deficits in Laboratory Mice", "type"=>"journal", "authors"=>[{"first_name"=>"Jia", "last_name"=>"Yao", "scopus_author_id"=>"57199375881"}, {"first_name"=>"Horacio O.", "last_name"=>"de la Iglesia", "scopus_author_id"=>"6602832047"}, {"first_name"=>"Sandra M.", "last_name"=>"Bajjalieh", "scopus_author_id"=>"6701346766"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "scopus"=>"2-s2.0-84880852195", "pui"=>"369437973", "doi"=>"10.1371/journal.pone.0068215", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "sgr"=>"84880852195", "pmid"=>"23894296"}, "id"=>"77ae2e0e-eb5a-3c1d-ac40-c673b05150b3", "abstract"=>"Neurons express two families of transporter-like proteins - Synaptic Vesicle protein 2 (SV2A, B, and C) and SV2-related proteins (SVOP and SVOPL). Both families share structural similarity with the Major Facilitator (MF) family of transporters. SV2 is present in all neurons and endocrine cells, consistent with it playing a key role in regulated exocytosis. Like SV2, SVOP is expressed in all brain regions, with highest levels in cerebellum, hindbrain and pineal gland. Furthermore, SVOP is expressed earlier in development than SV2 and is one of the neuronal proteins whose expression declines most during aging. Although SV2 is essential for survival, it is not required for development. Because significant levels of neurotransmission remain in the absence of SV2 it has been proposed that SVOP performs a function similar to that of SV2 that mitigates the phenotype of SV2 knockout mice. To test this, we generated SVOP knockout mice and SVOP/SV2A/SV2B triple knockout mice. Mice lacking SVOP are viable, fertile and phenotypically normal. Measures of neurotransmission and behaviors dependent on the cerebellum and pineal gland revealed no measurable phenotype. SVOP/SV2A/SV2B triple knockout mice did not display a phenotype more severe than mice harboring the SV2A/SV2B gene deletions. These findings support the interpretation that SVOP performs a unique, though subtle, function that is not necessary for survival under normal conditions.", "link"=>"http://www.mendeley.com/research/loss-sv2like-protein-svop-produces-apparent-deficits-laboratory-mice", "reader_count"=>19, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>1, "Researcher"=>6, "Student > Ph. D. Student"=>2, "Other"=>2, "Student > Master"=>1, "Student > Bachelor"=>4, "Professor"=>2}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>1, "Researcher"=>6, "Student > Ph. D. Student"=>2, "Other"=>2, "Student > Master"=>1, "Student > Bachelor"=>4, "Professor"=>2}, "reader_count_by_subject_area"=>{"Unspecified"=>2, "Biochemistry, Genetics and Molecular Biology"=>3, "Agricultural and Biological Sciences"=>7, "Medicine and Dentistry"=>3, "Neuroscience"=>4}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>3}, "Neuroscience"=>{"Neuroscience"=>4}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>7}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}, "Unspecified"=>{"Unspecified"=>2}}, "reader_count_by_country"=>{"United Kingdom"=>1, "Thailand"=>1}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1128936"], "description"=>"<p><b>A</b>, <b>B</b>) Shown are representative actograms and periodograms of wheel-running activity in wild-type (<b>A</b>) and SVOP knockout (<b>B</b>) mice under a 12 h : 12 h light/dark cycle and 12h: 12h dark/dark cycle. Left panels show representative actograms of two days’ activity plotted twice to demonstrate the circadian pattern. Right panels are periodograms that show the percent of behavioral variance (% V) accounted for by a circadian rhythm of <i>x</i> hours long. <b>C</b>) Calculated circadian period in constant darkness (DD) was similar for wild-type (+/+) and SVOP knockout (-/-) mice. The plot shows the mean ± SEM of the values calculated by periodogram analysis (two-tailed Student <i>t</i> test, <i>P</i> = 0.85, n = 6 for each genotype).</p>", "links"=>[], "tags"=>["svop", "knockout"], "article_id"=>753498, "categories"=>["Biological Sciences"], "users"=>["Jia Yao", "Horacio O. de la Iglesia", "Sandra M. Bajjalieh"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0068215.g007", "stats"=>{"downloads"=>1, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Circadian_activity_is_normal_in_SVOP_knockout_mice_/753498", "title"=>"Circadian activity is normal in SVOP knockout mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-07-24 01:41:13"}
  • {"files"=>["https://ndownloader.figshare.com/files/1128931"], "description"=>"<p>Neuronal cultures were generated and infected as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0068215#pone-0068215-g003\" target=\"_blank\">Figure <b>3</b></a>. <b>A)</b> Representative traces of single EPSCs. Neurons were held at −60 mV, and single EPSCs were evoked by depolarizing to +20mv for 1 ms. <b>B)</b> Average EPSC peak amplitudes of each experimental group. Data are presented as mean ± SEM. The numbers of cells analyzed are indicated within each column. There was no significant difference in the peak amplitude of EPSC between experimental groups (SVOP<sup>WT/WT</sup> - Cre, 4.03 ± 0.51 nA; SVOP<sup>WT/WT</sup> + Cre, 4.24 ± 0.58 nA, SVOP<sup>Flox/Flox</sup> -Cre 4.20 ± 0.60 nA and SVOP<sup>Flox/Flox</sup> + Cre 4.15 ± 0.58 nA, <i>p</i> = 0.99, two-way ANOVA). <b>C)</b> Representative traces of paired responses. Autaptic neurons were stimulated with two 1ms pulses separated by 45 ms. <b>D)</b> The paired-pulse ratio (PPR) (EPSC peak amplitude response 2/ response 1) was calculated for each cell. The graph shows the mean ± SEM for each experimental group. All experimental groups showed synaptic depression with a mean PPR of less than 1. The mean PPR across experimental groups was indistinguishable (<i>p</i> = 0.71, two-way ANOVA). <b>E)</b> Representative traces of EPSC in response to stimulus trains delivered at a frequency of 10 Hz. <b>F)</b> Shown are mean normalized EPSC amplitudes. Neurotransmission depressed at comparable rates in neurons from all experimental groups. Error bars represent the SEM for each point. The number of cells analyzed is indicated in parentheses. Data are from 10 different cultures. <b>G</b>, <b>H</b>) 20 Hz stimulus trains produced similar synaptic depression across groups.</p>", "links"=>[], "tags"=>["svop", "action-potential", "evoked", "transmitter"], "article_id"=>753493, "categories"=>["Biological Sciences"], "users"=>["Jia Yao", "Horacio O. de la Iglesia", "Sandra M. Bajjalieh"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0068215.g004", "stats"=>{"downloads"=>1, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Loss_of_SVOP_has_no_effect_on_action_potential_evoked_transmitter_release_or_short_term_plasticity_/753493", "title"=>"Loss of SVOP has no effect on action-potential evoked transmitter release or short-term plasticity.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-07-24 01:41:13"}
  • {"files"=>["https://ndownloader.figshare.com/files/1128930"], "description"=>"<p>Shown are graphs summarizing peak amplitude and frequency of mEPSC. Autaptic hippocampal neurons cultured from mice homozygous for Floxed SVOP genes or wild type littermate controls were infected with lenti virions encoding Cre-eGFP or eGFP. Neurons were analyzed in the whole-cell voltage-clamp configuration at DIV 14-20. Cells were selected for recording according to green fluorescence expression with preference given to brighter cells. Neurons were held at −60 mV, and mini EPSC were recorded in 5 min epochs. Graphs show the mean ± SEM. The number of cells analyzed is indicated within each column. Data are from three different cultures. <b>A)</b> Representative traces of mini EPSCs recorded from autaptic hippocampal neurons. <b>B)</b> Mean amplitudes of mEPSCs were unchanged across four experimental groups (SVOP<sup>WT/WT</sup> - Cre, 24.78 ± 1.56 pA; SVOP<sup>WT/WT</sup> + Cre, 24.74 ± 1.73 pA, SVOP<sup>Flox/Flox</sup> -Cre 26.13 ± 1.10 pA and SVOP<sup>Flox/Flox</sup> + Cre 26.28 ± 2.03 pA, <i>p</i> = 0.88, two-way ANOVA). <b>C)</b> Cumulative probability plots of mEPSC amplitude. The left panel shows the full amplitude range, the right panel is a plot of events with amplitudes < 80 pA. <b>D)</b> No differences in mEPSC frequency were observed across all experimental groups (SVOP<sup>WT/WT</sup> - Cre, 3.76 ± 0.83 Hz; SVOP<sup>WT/WT</sup> + Cre, 4.00 ± 0.77 Hz, SVOP<sup>Flox/Flox</sup> -Cre 4.95 ± 1.06 Hz and SVOP<sup>Flox/Flox</sup> + Cre 5.24 ± 1.58 Hz, <i>p</i> = 0.76, two-way ANOVA). <b>E)</b> Cumulative probability plots of mEPSC inter-event interval of 4 experimental groups. The left panel is a plot of the full inter-event interval range, the right panel of inter-event intervals <4000ms.</p>", "links"=>[], "tags"=>["svop", "spontaneous", "neurotransmitter"], "article_id"=>753492, "categories"=>["Biological Sciences"], "users"=>["Jia Yao", "Horacio O. de la Iglesia", "Sandra M. Bajjalieh"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0068215.g003", "stats"=>{"downloads"=>0, "page_views"=>16, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Loss_of_SVOP_does_not_affect_spontaneous_neurotransmitter_release_/753492", "title"=>"Loss of SVOP does not affect spontaneous neurotransmitter release.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-07-24 01:41:13"}
  • {"files"=>["https://ndownloader.figshare.com/files/1128935"], "description"=>"<p>Mice were tested for motor coordination on the rotating rod. Twelve SVOP knockout (-/-) mice and 12 SVOP wild type (+/+) littermates were included in the test. Shown are average fall latencies across 12 trials (3 trials per day over 4 consecutive days). Each data point represents the group mean± SEM. Performance between groups did not differ (Students <i>t</i> test, <i>p</i> values ranged from 0.17 to 0.98 across trials, SAS software analysis (GLIMMIX) generalized linear mixed analysis of intra-individual correlation over time, <i>P</i>=0.90). Therefore, loss of SVOP did not affect motor performance.</p>", "links"=>[], "tags"=>["svop"], "article_id"=>753497, "categories"=>["Biological Sciences"], "users"=>["Jia Yao", "Horacio O. de la Iglesia", "Sandra M. Bajjalieh"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0068215.g006", "stats"=>{"downloads"=>0, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Loss_of_SVOP_does_not_affect_motor_coordination_/753497", "title"=>"Loss of SVOP does not affect motor coordination.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-07-24 01:41:13"}
  • {"files"=>["https://ndownloader.figshare.com/files/1128933"], "description"=>"<div><p><b>A)</b> Shown are representative Western blot analyses of brain post nuclear supernatant isolated from littermate mice of the indicated genotype. The linear range was determined for each antibody used in Western blot analyses. The amount of protein loaded for each analysis fell within the linear range. Specifically, 5µg of protein were used for detecting SV2, α –adaptin, proton ATPase and VGlut1; 2.5µg of protein were used for detecting synaptophysin and synaptotagmin; 1.5µg of protein were used for detecting Vamp2 and clathrin heavy chain. The antibody used to detect SVOP is derived against a peptide in the amino terminus of SVOP.</p>\n\t\t\t\t\t\t<p><b>B)</b> Quantification of synaptic protein levels normalized to the amount of actin in the same lane. Brain tissue from 4 wild type, 6 heterozygous and 2 homozygous pups was analyzed. The histograms represent the mean value of expression normalized to wild type values in the same blot. Each dot represents a single datum. Loss of SVOP did not significantly alter expression levels of other synaptic vesicle proteins including Vamp2, the proton ATPase, synaptophysin, VGlut1, SV2, synaptotagmin as well as endocytic proteins such as α adaptin and clathrin heavy chain.</p></div>", "links"=>[], "tags"=>["svop", "synaptic"], "article_id"=>753495, "categories"=>["Biological Sciences"], "users"=>["Jia Yao", "Horacio O. de la Iglesia", "Sandra M. Bajjalieh"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0068215.g005", "stats"=>{"downloads"=>4, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Disruption_of_the_SVOP_gene_does_not_change_the_expression_of_other_synaptic_proteins_/753495", "title"=>"Disruption of the SVOP gene does not change the expression of other synaptic proteins.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-07-24 01:41:13"}
  • {"files"=>["https://ndownloader.figshare.com/files/1128929"], "description"=>"<p><b>A</b>) Shown are representative Western blots of protein expression in primary hippocampal neurons cultured from newborn pups homozygous for the targeted SVOP gene, or from wild type pups. Neurons were infected with lenti virions encoding Cre-eGFP or eGFP. The expression level of SVOP and the indicated proteins were detected by Western blot analysis. β3-tubulin, a neuron-specific isoform of tubulin, was probed for as a loading control. <b>B</b>) Plots of Western blot results from two independent experiments. Each dot represents one datum, and a horizontal line indicates the mean. The signals for SVOP, SV2 and synaptotagmin were normalized to β3-tubulin in the same lane and expressed as a percentage of the wild type - Cre lane in the same blot. SVOP was dramatically decreased in SVOP<sup>Flox/Flox</sup> neurons expressing Cre. Expression of the synaptic vesicle proteins synaptotagmin and SV2 was similar across conditions.</p>", "links"=>[], "tags"=>["recombinase", "disrupts", "svop", "neurons", "mice", "homozygous", "floxed"], "article_id"=>753491, "categories"=>["Biological Sciences"], "users"=>["Jia Yao", "Horacio O. de la Iglesia", "Sandra M. Bajjalieh"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0068215.g002", "stats"=>{"downloads"=>0, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Cre_recombinase_disrupts_SVOP_expression_in_neurons_from_mice_homozygous_for_the_floxed_SVOP_gene_/753491", "title"=>"Cre recombinase disrupts SVOP expression in neurons from mice homozygous for the floxed SVOP gene.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-07-24 01:41:13"}
  • {"files"=>["https://ndownloader.figshare.com/files/1128928"], "description"=>"<p>Shown is the strategy used to generate mice lacking SVOP. <b>A)</b> An 11.4kb genomic DNA containing exons 2-5 of the SVOP gene was used for generating a targeting construct. <b>B)</b> A targeting construct was generated in which exons 2 and 3 were flanked by Cre recombinase recognition (<i>loxP</i>) sites. A cDNA encoding neomycin resistance protein was included to allow screening of embryonic stem cells. To allow removal of the neomycin resistant gene, it was flanked by Flipper recombinase recognition (Frt) sites. <b>C)</b> Map of the targeted gene generated by homologous recombination. <b>D)</b> Upon Cre recombinase-induced excision, exons 2 and 3 are removed. This results in a truncated protein after the codon encoding a.a.18.</p>", "links"=>[], "tags"=>["disruption", "svop"], "article_id"=>753490, "categories"=>["Biological Sciences"], "users"=>["Jia Yao", "Horacio O. de la Iglesia", "Sandra M. Bajjalieh"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0068215.g001", "stats"=>{"downloads"=>0, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Targeted_disruption_of_the_SVOP_gene_/753490", "title"=>"Targeted disruption of the SVOP gene.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-07-24 01:41:13"}

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Relative Metric

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