Vivo-Morpholinos Induced Transient Knockdown of Physical Activity Related Proteins
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{"title"=>"Vivo-Morpholinos Induced Transient Knockdown of Physical Activity Related Proteins", "type"=>"journal", "authors"=>[{"first_name"=>"David P.", "last_name"=>"Ferguson", "scopus_author_id"=>"37030766300"}, {"first_name"=>"Emily E.", "last_name"=>"Schmitt", "scopus_author_id"=>"55955395300"}, {"first_name"=>"J. Timothy", "last_name"=>"Lightfoot", "scopus_author_id"=>"7006575856"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"sgr"=>"84876421893", "doi"=>"10.1371/journal.pone.0061472", "pui"=>"368766944", "pmid"=>"23630592", "scopus"=>"2-s2.0-84876421893", "issn"=>"19326203"}, "id"=>"ac20a8fd-2c6a-302a-bc66-8d18b08a8cb9", "abstract"=>"Physical activity is associated with disease prevention and overall wellbeing. Additionally there has been evidence that physical activity level is a result of genetic influence. However, there has not been a reliable method to silence candidate genes in vivo to determine causal mechanisms of physical activity regulation. Vivo-morpholinos are a potential method to transiently silence specific genes. Thus, the aim of this study was to validate the use of Vivo-morpholinos in a mouse model for voluntary physical activity with several sub-objectives. We observed that Vivo-morpholinos achieved between 60-97% knockdown of Drd1-, Vmat2-, and Glut4-protein in skeletal muscle, the delivery moiety of Vivo-morpholinos (scramble) did not influence physical activity and that a cocktail of multiple Vivo-morpholinos can be given in a single treatment to achieve protein knockdown of two different targeted proteins in skeletal muscle simultaneously. Knocking down Drd1, Vmat2, or Glut4 protein in skeletal muscle did not affect physical activity. Vivo-morpholinos injected intravenously alone did not significantly knockdown Vmat2-protein expression in the brain (p = 0.28). However, the use of a bradykinin analog to increase blood-brain-barrier permeability in conjunction with the Vivo-morpholinos significantly (p = 0.0001) decreased Vmat2-protein in the brain with a corresponding later over-expression of Vmat2 coincident with a significant (p = 0.0016) increase in physical activity. We conclude that Vivo-morpholinos can be a valuable tool in determining causal gene-phenotype relationships in whole animal models.", "link"=>"http://www.mendeley.com/research/vivomorpholinos-induced-transient-knockdown-physical-activity-related-proteins", "reader_count"=>25, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>1, "Researcher"=>3, "Student > Ph. D. Student"=>7, "Other"=>2, "Student > Master"=>5, "Student > Bachelor"=>6, "Professor"=>1}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>1, "Researcher"=>3, "Student > Ph. D. Student"=>7, "Other"=>2, "Student > Master"=>5, "Student > Bachelor"=>6, "Professor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>1, "Environmental Science"=>1, "Nursing and Health Professions"=>1, "Biochemistry, Genetics and Molecular Biology"=>4, "Agricultural and Biological Sciences"=>12, "Medicine and Dentistry"=>3, "Sports and Recreations"=>1, "Earth and Planetary Sciences"=>2}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>3}, "Sports and Recreations"=>{"Sports and Recreations"=>1}, "Earth and Planetary Sciences"=>{"Earth and Planetary Sciences"=>2}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>12}, "Nursing and Health Professions"=>{"Nursing and Health Professions"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>4}, "Unspecified"=>{"Unspecified"=>1}, "Environmental Science"=>{"Environmental Science"=>1}}, "reader_count_by_country"=>{"United States"=>1}, "group_count"=>2}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1030954"], "description"=>"<p>Mean ± standard deviation of western blot optical band density and a representative western blot comparing Saline, Scramble and Vivo-morpholino treated animals for Vmat2 protein expression in the nucleus accumbens of the brain. There was no difference between the treatment groups at four (p = 0.87, Panel A) or eleven (p = 0.14, Panel B) days post treatment.</p>", "links"=>[], "tags"=>["Anatomy and physiology", "genetics", "gene expression", "RNA interference", "Animal genetics", "Gene function", "genomics", "Functional genomics", "Model organisms", "Animal models", "mouse", "Zoology", "Animal physiology", "vmat2", "nucleus"], "article_id"=>687515, "categories"=>["Biological Sciences"], "users"=>["David P. Ferguson", "Emily E. Schmitt", "J. Timothy Lightfoot"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0061472.g001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Experiment_1_Vmat2_protein_level_in_the_nucleus_accumbens_/687515", "title"=>"Experiment 1, Vmat2 protein level in the nucleus accumbens.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-04-22 02:05:15"}
  • {"files"=>["https://ndownloader.figshare.com/files/1030955"], "description"=>"<p>Mean ± standard deviation of western blot optical band density and a representative western blot comparing Saline, Scramble and Vivo-morpholino treated animals for Vmat2 protein expression in the soleus. Panel A shows Vmat2 expression at four days post-last injection. There was a significant decrease (p = 0.04) in Vivo-morpholino treated animals compared to Saline and Scramble treated animals in Vmat2 expression at four days post injection. Panel B shows Vmat2 expression at eleven days post-last injection. There was a significant increase (p = 0.03) in Vivo-morpholino treated animals compared to Saline and Scramble treated animals in Vmat2 expression at eleven days post injection.</p>", "links"=>[], "tags"=>["Anatomy and physiology", "genetics", "gene expression", "RNA interference", "Animal genetics", "Gene function", "genomics", "Functional genomics", "Model organisms", "Animal models", "mouse", "Zoology", "Animal physiology", "vmat2"], "article_id"=>687516, "categories"=>["Biological Sciences"], "users"=>["David P. Ferguson", "Emily E. Schmitt", "J. Timothy Lightfoot"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0061472.g002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Experiment_1_Vmat2_protein_level_in_the_soleus_/687516", "title"=>"Experiment 1, Vmat2 protein level in the soleus.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-04-22 02:05:16"}
  • {"files"=>["https://ndownloader.figshare.com/files/1030956"], "description"=>"<p>Average daily distance ran for the baseline, injection, and recovery week for animals treated with saline, Vivo-morpholino vehicle only (scramble) and the Vivo-morpholino targeting <i>Vmat2</i> (Vmat2). Vmat2 group significantly (*p = 0.001) increased activity in the recovery week compared to the baseline and injection week. There was no difference in physical activity (p>0.05) in the scramble or saline treated animals for baseline, injection, and recovery weeks.</p>", "links"=>[], "tags"=>["Anatomy and physiology", "genetics", "gene expression", "RNA interference", "Animal genetics", "Gene function", "genomics", "Functional genomics", "Model organisms", "Animal models", "mouse", "Zoology", "Animal physiology"], "article_id"=>687517, "categories"=>["Biological Sciences"], "users"=>["David P. Ferguson", "Emily E. Schmitt", "J. Timothy Lightfoot"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0061472.g003"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Experiment_1_mouse_wheel_running_/687517", "title"=>"Experiment 1, mouse wheel running.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-04-22 02:05:17"}
  • {"files"=>["https://ndownloader.figshare.com/files/1030957"], "description"=>"<p>Panels A, B, and C represent soleus samples while panels D, E, and F represent nucleus accumbens samples. Panel A represents western blot data probing for Vmat2 in the soleus for both unlocked wheel and locked wheel groups. Panel B is the optical density of the individual western blot bands from panel A. A nonlinear regression was run on the data in panel B which generated an R<sup>2</sup> = 0.65 for the animals on unlocked wheels and an R<sup>2</sup> = 0.55 for animals on locked wheels. Given there was no difference (p = 0.42) in the slope or y intercept of the unlocked wheel and locked wheel groups, the data were pooled at each Day (Panel C). Panel C shows that there was a significant knockdown (<sup>+</sup>p = 0.001) in Vmat2 on days 2–6 and a significant (<sup>*</sup>p = 0.001) over expression in Vmat2 on day 9 in the soleus. The same methodologies were applied to Panels D, E, and F for the nucleus accumbens. The nonlinear regression resulted in R<sup>2</sup> = 0.99 for the unlocked wheel group and R<sup>2</sup> = 0.99 for the locked wheel group, and there was no difference (p = 0.66) in the slope and y intercepts of the lines in panel E. Additionally Vmat2 was only significantly knocked down on Days 5 and 6 (<sup>+</sup>p = 0.001) with a significant over expression (<sup>*</sup>p = 0.001) on Day 11. The mouse in the locked wheel group representing data for Day 11 was removed from analysis because it developed malocclusion in the front incisors and became malnourished, thus altering physical activity level.</p>", "links"=>[], "tags"=>["Anatomy and physiology", "genetics", "gene expression", "RNA interference", "Animal genetics", "Gene function", "genomics", "Functional genomics", "Model organisms", "Animal models", "mouse", "Zoology", "Animal physiology", "washout", "vivo-morpholinos", "targeting"], "article_id"=>687518, "categories"=>["Biological Sciences"], "users"=>["David P. Ferguson", "Emily E. Schmitt", "J. Timothy Lightfoot"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0061472.g004"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Experiment_2_Evaluation_of_the_washout_time_course_of_Vivo_morpholinos_targeting_Vmat2_/687518", "title"=>"Experiment 2, Evaluation of the washout time course of Vivo-morpholinos targeting <i>Vmat2</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-04-22 02:05:18"}
  • {"files"=>["https://ndownloader.figshare.com/files/1030959"], "description"=>"<p>Western blot analysis probing for α-synuclein to confirm the signal for the over expression of Vmat2 protein seen in the nucleus accumbens. There was an increase in α-synuclein at days 2 and 7–10. α-synuclein is an indicator of <i>Vmat2</i> transcription thus when Vmat2 was knocked down by the Vivo-morpholino there was an increased stimulus placed on the cell to transcribe more <i>Vmat2</i>.</p>", "links"=>[], "tags"=>["Anatomy and physiology", "genetics", "gene expression", "RNA interference", "Animal genetics", "Gene function", "genomics", "Functional genomics", "Model organisms", "Animal models", "mouse", "Zoology", "Animal physiology", "nucleus"], "article_id"=>687519, "categories"=>["Biological Sciences"], "users"=>["David P. Ferguson", "Emily E. Schmitt", "J. Timothy Lightfoot"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0061472.g005"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Experiment_2_945_synuclein_protein_level_in_nucleus_accumbens_/687519", "title"=>"Experiment 2, α-synuclein protein level in nucleus accumbens.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-04-22 02:05:19"}
  • {"files"=>["https://ndownloader.figshare.com/files/1030960"], "description"=>"<p>Average daily distance ran for animals treated with Vivo-morpholino targeting <i>Vmat2</i> for the baseline week, injection week (Days 1–5), and recovery week (Days 6–10) of washout protocol. Physical activity was significantly (*p = 0.0016) increased in Days 6–10.</p>", "links"=>[], "tags"=>["Anatomy and physiology", "genetics", "gene expression", "RNA interference", "Animal genetics", "Gene function", "genomics", "Functional genomics", "Model organisms", "Animal models", "mouse", "Zoology", "Animal physiology"], "article_id"=>687520, "categories"=>["Biological Sciences"], "users"=>["David P. Ferguson", "Emily E. Schmitt", "J. Timothy Lightfoot"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0061472.g006"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Experiment_2_mouse_wheel_running_/687520", "title"=>"Experiment 2, mouse wheel running.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-04-22 02:05:20"}
  • {"files"=>["https://ndownloader.figshare.com/files/1030962"], "description"=>"<p>Western blot and mean ± standard deviation of optical densities of soleus tissue that underwent the Vivo-morpholino cocktail treatment. Panel A represents four days post treatment of Drd1 and Glut4 expression comparing controls to Vivo-morpholino cocktail treated animals. There was a significant knockdown for both Drd1 (p = 0.01) and Glut4 (p = 0.04). Panel B represents eleven days post treatment of Vivo-morpholino cocktail. There was no difference between control and Drd1 (p = 0.79) and Glut4 (p = 0.58) expression.</p>", "links"=>[], "tags"=>["Anatomy and physiology", "genetics", "gene expression", "RNA interference", "Animal genetics", "Gene function", "genomics", "Functional genomics", "Model organisms", "Animal models", "mouse", "Zoology", "Animal physiology", "drd1", "glut4"], "article_id"=>687521, "categories"=>["Biological Sciences"], "users"=>["David P. Ferguson", "Emily E. Schmitt", "J. Timothy Lightfoot"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0061472.g007"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Experiment_3_Drd1_and_Glut4_protein_level_/687521", "title"=>"Experiment 3, Drd1 and Glut4 protein level.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-04-22 02:05:21"}
  • {"files"=>["https://ndownloader.figshare.com/files/1030963"], "description"=>"<p>Average daily distance ran for baseline, injection, and recovery week for animals treated saline (control) and the morpholino cocktail targeting Drd1 and Glut4 (Vivo-morpholino). There was no difference (p = 0.15) between the activity of the control group and the morpholino group.</p>", "links"=>[], "tags"=>["Anatomy and physiology", "genetics", "gene expression", "RNA interference", "Animal genetics", "Gene function", "genomics", "Functional genomics", "Model organisms", "Animal models", "mouse", "Zoology", "Animal physiology"], "article_id"=>687522, "categories"=>["Biological Sciences"], "users"=>["David P. Ferguson", "Emily E. Schmitt", "J. Timothy Lightfoot"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0061472.g008"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Experiment_3_mouse_wheel_running_/687522", "title"=>"Experiment 3, mouse wheel running.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-04-22 02:05:22"}
  • {"files"=>["https://ndownloader.figshare.com/files/1030964"], "description"=>"<p>Western blot and mean ± standard deviation of optical densities of soleus tissue that underwent the Vivo-morpholino cocktail treatment. Panel A represents four days post treatment of Drd5 and Glut1 protein expression comparing controls to Vivo-morpholino cocktail treated animals. There was not a significant knockdown for either Drd5 (p = 0.22) or Glut1 (p = 0.56). Panel B represents eleven days post treatment of Vivo-morpholino cocktail. There was no difference between control and Drd5 (p = 0.93) and Glut1 (p = 0.36) expression.</p>", "links"=>[], "tags"=>["Anatomy and physiology", "genetics", "gene expression", "RNA interference", "Animal genetics", "Gene function", "genomics", "Functional genomics", "Model organisms", "Animal models", "mouse", "Zoology", "Animal physiology", "drd5", "glut1"], "article_id"=>687523, "categories"=>["Biological Sciences"], "users"=>["David P. Ferguson", "Emily E. Schmitt", "J. Timothy Lightfoot"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0061472.g009"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Experiment_3_Drd5_and_Glut1_protein_level_/687523", "title"=>"Experiment 3, Drd5 and Glut1 protein level.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-04-22 02:05:23"}

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Relative Metric

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