Down-Regulation of KCC2 Expression and Phosphorylation in Motoneurons, and Increases the Number of in Primary Afferent Projections to Motoneurons in Mice with Post-Stroke Spasticity
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{"title"=>"Down-regulation of KCC2 expression and phosphorylation in motoneurons, and increases the number of in primary afferent projections to motoneurons in mice with post-stroke spasticity", "type"=>"journal", "authors"=>[{"first_name"=>"Takuya", "last_name"=>"Toda", "scopus_author_id"=>"56348816900"}, {"first_name"=>"Kazuto", "last_name"=>"Ishida", "scopus_author_id"=>"57190215555"}, {"first_name"=>"Hiroshi", "last_name"=>"Kiyama", "scopus_author_id"=>"10141653000"}, {"first_name"=>"Toshihide", "last_name"=>"Yamashita", "scopus_author_id"=>"35479670200"}, {"first_name"=>"Sachiko", "last_name"=>"Lee", "scopus_author_id"=>"40261932500"}], "year"=>2014, "source"=>"PLoS ONE", "identifiers"=>{"sgr"=>"84919941044", "pui"=>"601081443", "doi"=>"10.1371/journal.pone.0114328", "scopus"=>"2-s2.0-84919941044", "issn"=>"19326203"}, "id"=>"af7c04e0-83a9-3a43-a88f-512fcca993e4", "abstract"=>"© 2014 Toda et al.Spasticity obstructs motor function recovery post-stroke, and has been reported to occur in spinal cord injury and electrophysiological studies. The purpose of the present study was to assess spinal cord circuit spasticity in post-stroke mice. At 3, 7, 21, and 42 d after photothrombotic ischemic cortical injury in C57BL/6J mice, we observed decreased rate-dependent depression (RDD) of the Hoffmann reflex (H reflex) in the affected forelimb of mice compared with the limbs of sham mice and the non-affected forelimb. This finding suggests a hyper-excitable stretch reflex in the affected forelimb. We then performed immunohistochemical and western blot analyses to examine the expression of the potassium-chloride cotransporter 2 (KCC2) and phosphorylation of the KCC2 serine residue, 940 (S940), since this is the main chloride extruder that affects neuronal excitability. We also performed immunohistochemical analyses on the number of vesicular glutamate transporter 1 (vGluT1)-positive boutons to count the number of Ia afferent fibers that connect to motoneurons. Western bolts revealed that, compared with sham mice, experimental mice had significantly reduced KCC2 expression at 7 d post-stroke, and dephosphorylated S940 at 3 and 7 d post-stroke in motoneuron plasma membranes. We also observed a lower density of KCC2-positive areas in the plasma membrane of motoneurons at 3 and 7 d post-stroke. However, western blot and immunohistochemical analyses revealed that there were no differences between groups 21 and 42 d post-stroke, respectively. In addition, at 7 and 42 d post-stroke, experimental mice exhibited a significant increase in vGluT1 boutons compared with sham mice. Our findings suggest that both the down-regulation of KCC2 and increases in Ia afferent fibers are involved in post-stroke spasticity.", "link"=>"http://www.mendeley.com/research/downregulation-kcc2-expression-phosphorylation-motoneurons-increases-number-primary-afferent-project", "reader_count"=>24, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>1, "Librarian"=>1, "Researcher"=>5, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>10, "Student > Postgraduate"=>1, "Student > Bachelor"=>2, "Lecturer > Senior Lecturer"=>1, "Professor"=>1}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>1, "Librarian"=>1, "Researcher"=>5, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>10, "Student > Postgraduate"=>1, "Student > Bachelor"=>2, "Lecturer > Senior Lecturer"=>1, "Professor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>1, "Biochemistry, Genetics and Molecular Biology"=>2, "Agricultural and Biological Sciences"=>11, "Medicine and Dentistry"=>2, "Neuroscience"=>5, "Philosophy"=>1, "Psychology"=>1, "Computer Science"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>2}, "Neuroscience"=>{"Neuroscience"=>5}, "Psychology"=>{"Psychology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>11}, "Computer Science"=>{"Computer Science"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>2}, "Unspecified"=>{"Unspecified"=>1}, "Philosophy"=>{"Philosophy"=>1}}, "reader_count_by_country"=>{"Finland"=>1}, "group_count"=>2}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/4341844"], "description"=>"<p>A: RDDs of the H reflexes at baseline (pre-stroke). B: Changes in RDDs of the H reflex 7 d post-stroke in sham and stroke animals. Data are presented as percentages relative to the mean amplitude at 0.1 Hz in the same test. C: Changes in RDDs of the H reflex with 5 Hz stimulation on the affected and non-affected sides in stroke and sham mice. Error bars indicate S.E.M. One-way ANOVA with post hoc Tukey-Kramer tests, ##p<0.01, between stroke-affected and sham-affected sides, *<i>p</i><0.05 and **<i>p</i><0.01, between stroke-affected and sham non-affected sides, †<i>p</i><0.05 and ††<i>p</i><0.01, between stroke-affected and stroke non-affected, §<i>p</i><0.05, between stroke non-affected and sham-affected sides.</p>", "links"=>[], "tags"=>["Primary Afferent Projections", "immunohistochemical analyses", "vGluT 1 boutons", "sham mice", "cord circuit spasticity", "glutamate transporter 1", "forelimb", "KCC 2 Expression", "KCC 2 serine residue", "reflex", "KCC 2 expression", "RDD", "C 57BL mice", "motoneuron plasma membranes", "dephosphorylated S 940"], "article_id"=>2678128, "categories"=>["Biophysics", "Biochemistry", "Microbiology", "Cell Biology", "Molecular Biology", "Neuroscience", "Physiology", "Developmental Biology", "Mental Health", "Virology"], "users"=>["Takuya Toda", "Kazuto Ishida", "Hiroshi Kiyama", "Toshihide Yamashita", "Sachiko Lee"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0114328.g003", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Changes_in_H_reflex_RDDs_after_stroke_/2678128", "title"=>"Changes in H reflex RDDs after stroke.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-12-29 15:57:19"}
  • {"files"=>["https://ndownloader.figshare.com/files/4341853"], "description"=>"<p>A: Immunofluorescence photomicrographs captured with confocal microscopy showing individual motoneurons labeled with ChAT in magenta, KCC2 in green, and DAPI in blue. Merged image was reconstructed to three dimensional (3D) Z stack images. Arrowheads show the discontinuous labeling of KCC2 in the plasma membrane. Scale bar = 20 “Insert>Symbols” µ m. B-D: Semi-quantification of the density of KCC2 labeling in plasma membranes of spinal motoneurons. Labeled KCC2 is shown as a percentage of the pixel surface per somatic perimeter in the sham-affected side at 3, 7, and 42 d post-stroke, respectively.</p>", "links"=>[], "tags"=>["Primary Afferent Projections", "immunohistochemical analyses", "vGluT 1 boutons", "sham mice", "cord circuit spasticity", "glutamate transporter 1", "forelimb", "KCC 2 Expression", "KCC 2 serine residue", "reflex", "KCC 2 expression", "RDD", "C 57BL mice", "motoneuron plasma membranes", "dephosphorylated S 940"], "article_id"=>2678137, "categories"=>["Biophysics", "Biochemistry", "Microbiology", "Cell Biology", "Molecular Biology", "Neuroscience", "Physiology", "Developmental Biology", "Mental Health", "Virology"], "users"=>["Takuya Toda", "Kazuto Ishida", "Hiroshi Kiyama", "Toshihide Yamashita", "Sachiko Lee"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0114328.g004", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Down_regulation_of_KCC2_levels_using_immunohistochemical_analysis_in_motoneuron_plasma_membrane_after_stroke_/2678137", "title"=>"Down-regulation of KCC2 levels using immunohistochemical analysis in motoneuron plasma membrane after stroke.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-12-29 15:57:19"}
  • {"files"=>["https://ndownloader.figshare.com/files/4341838"], "description"=>"<p>A: An illustration of the H reflex in the spinal cord circuit. The M wave is produced by orthodromically transmitting stimulation to motor fibers. The H reflex is a monosynaptic reflex that is transmitted via Ia afferent fibers. B and C: Evoked electromyograms measured in sham mice (B) and on the affected forelimb of stroke mice (C) at 7 d post-stroke. Repeated stimulation at high frequency (5 Hz) decreased the amplitude of the H reflex in sham mice (B). The amplitude of the H reflex on the affected forelimb of stroke mice was not decreased following 5 Hz stimulation (C).</p>", "links"=>[], "tags"=>["Primary Afferent Projections", "immunohistochemical analyses", "vGluT 1 boutons", "sham mice", "cord circuit spasticity", "glutamate transporter 1", "forelimb", "KCC 2 Expression", "KCC 2 serine residue", "reflex", "KCC 2 expression", "RDD", "C 57BL mice", "motoneuron plasma membranes", "dephosphorylated S 940"], "article_id"=>2678122, "categories"=>["Biophysics", "Biochemistry", "Microbiology", "Cell Biology", "Molecular Biology", "Neuroscience", "Physiology", "Developmental Biology", "Mental Health", "Virology"], "users"=>["Takuya Toda", "Kazuto Ishida", "Hiroshi Kiyama", "Toshihide Yamashita", "Sachiko Lee"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0114328.g002", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/The_RDD_of_the_H_reflex_/2678122", "title"=>"The RDD of the H reflex.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-12-29 15:57:19"}
  • {"files"=>["https://ndownloader.figshare.com/files/4341865"], "description"=>"<p>A: Dual labeling of vGluT1 (green) and ChAT (magenta) at 3, 7, and 42 d after stroke. Arrowheads show vGuT1-positive boutons contacting motoneuron somata and the arrows show non-counted boutons because the boutons did not contact the somata. Scale bar = 20 “Insert>Symbols” µ m. B-D: Quantification of the number of vGluT1-positive boutons on plasma membranes of spinal motoneurons in sham and stroke mice at 3, 7, and 42 d after stroke. Error bars on graphs represent S.E.M. One-way ANOVA with post hoc Tukey-Kramer test, **p<0.01.</p>", "links"=>[], "tags"=>["Primary Afferent Projections", "immunohistochemical analyses", "vGluT 1 boutons", "sham mice", "cord circuit spasticity", "glutamate transporter 1", "forelimb", "KCC 2 Expression", "KCC 2 serine residue", "reflex", "KCC 2 expression", "RDD", "C 57BL mice", "motoneuron plasma membranes", "dephosphorylated S 940"], "article_id"=>2678149, "categories"=>["Biophysics", "Biochemistry", "Microbiology", "Cell Biology", "Molecular Biology", "Neuroscience", "Physiology", "Developmental Biology", "Mental Health", "Virology"], "users"=>["Takuya Toda", "Kazuto Ishida", "Hiroshi Kiyama", "Toshihide Yamashita", "Sachiko Lee"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0114328.g006", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/The_number_of_vGluT1_positive_boutons_on_motoneurons_after_stroke_/2678149", "title"=>"The number of vGluT1-positive boutons on motoneurons after stroke.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-12-29 15:57:19"}
  • {"files"=>["https://ndownloader.figshare.com/files/4341832"], "description"=>"<p>A: Picture of a lesion at 7 d after stroke. B: A tracing of lesion areas (7 d post-stroke, n = 6). C: Nissl staining of coronal sections of the brain at 7 d post-stroke. The left rostral and caudal forelimb motor cortex show specific injuries. The mean injury volume was 5.13±1.23 mm<sup>3</sup> (n = 6).</p>", "links"=>[], "tags"=>["Primary Afferent Projections", "immunohistochemical analyses", "vGluT 1 boutons", "sham mice", "cord circuit spasticity", "glutamate transporter 1", "forelimb", "KCC 2 Expression", "KCC 2 serine residue", "reflex", "KCC 2 expression", "RDD", "C 57BL mice", "motoneuron plasma membranes", "dephosphorylated S 940"], "article_id"=>2678116, "categories"=>["Biophysics", "Biochemistry", "Microbiology", "Cell Biology", "Molecular Biology", "Neuroscience", "Physiology", "Developmental Biology", "Mental Health", "Virology"], "users"=>["Takuya Toda", "Kazuto Ishida", "Hiroshi Kiyama", "Toshihide Yamashita", "Sachiko Lee"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0114328.g001", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Photothrombotic_stroke_lesion_in_the_left_rostral_and_caudal_forelimb_motor_cortex_/2678116", "title"=>"Photothrombotic stroke lesion in the left rostral and caudal forelimb motor cortex.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-12-29 15:57:19"}
  • {"files"=>["https://ndownloader.figshare.com/files/4341859"], "description"=>"<p>A, B, and C: At 3, 7, and 21 d post-stroke, bands of KCC2, phosphorylated S940, NKCC1, and tubulin. D-L: Quantification analysis of KCC2, phosphorylated S940, NKCC1levels at 3, 7, and 21 d after stroke. White bars are sham animals, and black bars are stroke animals. Data are presented as relative ratio to the mean sham non-affected and as the mean ± S.E.M. One-way ANOVA post hoc test Tukey-Kramer tests: *p<0.05, **p<0.01.</p>", "links"=>[], "tags"=>["Primary Afferent Projections", "immunohistochemical analyses", "vGluT 1 boutons", "sham mice", "cord circuit spasticity", "glutamate transporter 1", "forelimb", "KCC 2 Expression", "KCC 2 serine residue", "reflex", "KCC 2 expression", "RDD", "C 57BL mice", "motoneuron plasma membranes", "dephosphorylated S 940"], "article_id"=>2678143, "categories"=>["Biophysics", "Biochemistry", "Microbiology", "Cell Biology", "Molecular Biology", "Neuroscience", "Physiology", "Developmental Biology", "Mental Health", "Virology"], "users"=>["Takuya Toda", "Kazuto Ishida", "Hiroshi Kiyama", "Toshihide Yamashita", "Sachiko Lee"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0114328.g005", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/KCC2_S940_phosphorylation_in_KCC2_and_NKCC1expression_levels_assessed_by_western_blot_analysis_in_membrane_enriched_fractions_of_the_ventral_horn_/2678143", "title"=>"KCC2, S940 phosphorylation in KCC2, and NKCC1expression levels assessed by western blot analysis in membrane-enriched fractions of the ventral horn.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-12-29 15:57:19"}

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