Establishment of a Knock-In Mouse Model with the SLC26A4 c.919-2A>G Mutation and Characterization of Its Pathology
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{"title"=>"Establishment of a knock-in mouse model with the SLC26A4 c.919-2a>G mutation and characterization of its pathology", "type"=>"journal", "authors"=>[{"first_name"=>"Ying Chang", "last_name"=>"Lu", "scopus_author_id"=>"35174929400"}, {"first_name"=>"Chen Chi", "last_name"=>"Wu", "scopus_author_id"=>"8451843700"}, {"first_name"=>"Wen Sheng", "last_name"=>"Shen", "scopus_author_id"=>"48662933900"}, {"first_name"=>"Ting Hua", "last_name"=>"Yang", "scopus_author_id"=>"24473916900"}, {"first_name"=>"Te Huei", "last_name"=>"Yeh", "scopus_author_id"=>"7201658033"}, {"first_name"=>"Pei Jer", "last_name"=>"Chen", "scopus_author_id"=>"7408354514"}, {"first_name"=>"I. Shing", "last_name"=>"Yu", "scopus_author_id"=>"7102120396"}, {"first_name"=>"Shu Wha", "last_name"=>"Lin", "scopus_author_id"=>"16201167800"}, {"first_name"=>"Jau Min", "last_name"=>"Wong", "scopus_author_id"=>"7404431489"}, {"first_name"=>"Qing", "last_name"=>"Chang", "scopus_author_id"=>"7101731419"}, {"first_name"=>"Xi", "last_name"=>"Lin", "scopus_author_id"=>"25028195900"}, {"first_name"=>"Chuan Jen", "last_name"=>"Hsu", "scopus_author_id"=>"7404946816"}], "year"=>2011, "source"=>"PLoS ONE", "identifiers"=>{"isbn"=>"1932-6203 (Electronic)\r1932-6203 (Linking)", "issn"=>"19326203", "pui"=>"362190910", "sgr"=>"79960650143", "pmid"=>"21811566", "scopus"=>"2-s2.0-79960650143", "doi"=>"10.1371/journal.pone.0022150"}, "id"=>"03a5b6ef-d6ed-3eea-9f54-6835d1b0ab30", "abstract"=>"Recessive mutations in the SLC26A4 gene are a common cause of hereditary hearing impairment worldwide. Previous studies have demonstrated that different SLC26A4 mutations may have different pathogenetic mechanisms. In the present study, we established a knock-in mouse model (i.e., Slc26a4(tm1Dontuh/tm1Dontuh) mice) homozygous for the c.919-2A>G mutation, which is a common mutation in East Asians. Mice were then subjected to audiologic assessment, a battery of vestibular evaluations, and inner ear morphological studies. All Slc26a4(tm1Dontuh/tm1Dontuh) mice revealed profound hearing loss, whereas 46% mice demonstrated pronounced head tilting and circling behaviors. There was a significant difference in the vestibular performance between wild-type and Slc26a4(tm1Dontuh/tm1Dontuh) mice, especially those exhibiting circling behavior. Inner ear morphological examination of Slc26a4(tm1Dontuh/tm1Dontuh) mice revealed an enlarged endolymphatic duct, vestibular aqueduct and sac, atrophy of stria vascularis, deformity of otoconia in the vestibular organs, consistent degeneration of cochlear hair cells, and variable degeneration of vestibular hair cells. Audiologic and inner ear morphological features of Slc26a4(tm1Dontuh/tm1Dontuh) mice were reminiscent of those observed in humans. These features were also similar to those previously reported in both knock-out Slc26a4(-/-) mice and Slc26a4(loop/loop) mice with the Slc26a4 p.S408F mutation, albeit the severity of vestibular hair cell degeneration appeared different among the three mouse strains.", "link"=>"http://www.mendeley.com/research/establishment-knockin-mouse-model-slc26a4-c9192ag-mutation-characterization-pathology", "reader_count"=>6, "reader_count_by_academic_status"=>{"Student > Ph. D. Student"=>2, "Student > Postgraduate"=>1, "Other"=>1, "Student > Bachelor"=>1, "Professor"=>1}, "reader_count_by_user_role"=>{"Student > Ph. D. Student"=>2, "Student > Postgraduate"=>1, "Other"=>1, "Student > Bachelor"=>1, "Professor"=>1}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>2, "Mathematics"=>1, "Agricultural and Biological Sciences"=>2, "Medicine and Dentistry"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>2}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>2}, "Mathematics"=>{"Mathematics"=>1}}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/754668"], "description"=>"<p><i>Slc26a4<sup>tm1Dontuh/tm1Dontuh</sup></i> mice revealed a decreased amount of otoconia in the saccule and utricle (B & D, arrowhead), formation of “giant otoconia” in the utricle (D & H), ectopic otoconia in the semicircular canal (F), as well as a wide spectrum of vestibular hair cell loss and degeneration (arrows) at 8 weeks of age. Scanning electron microscopy reveals the markedly enlarged otoconia at saccule and utricle in <i>Slc26a4<sup>tm1Dontuh/tm1Dontuh</sup></i> mice (H) as compared with wild-type (G). Degeneration of vestibular hair cells in <i>Slc26a4<sup>tm1Dontuh/tm1Dontuh</sup></i> mice was also revealed by fluorescence confocal microscopy (L). J, L: magnification of boxes J and L from figures I and K, respectively. EO, ectopic otoconia; GO, giant otoconia. Bar = 50 µm (A–F, I & K), and 10 µm (G, H, J & L).</p>", "links"=>[], "tags"=>["vestibular", "morphology", "mice", "semicircular", "utricular"], "article_id"=>425022, "categories"=>["Neuroscience", "Genetics"], "users"=>["Ying-Chang Lu", "Chen-Chi Wu", "Wen-Sheng Shen", "Ting-Hua Yang", "Te-Huei Yeh", "Pei-Jer Chen", "I-Shing Yu", "Shu-Wha Lin", "Jau-Min Wong", "Qing Chang", "Xi Lin", "Chuan-Jen Hsu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0022150.g007", "stats"=>{"downloads"=>3, "page_views"=>95, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Comparison_of_vestibular_morphology_between_Slc26a4_mice_A_saccule_C_utricle_E_semicircular_canal_G_utricular_macula_and_Slc26a4_tm1Dontuh_tm1Dontuh_mice_B_saccule_D_utricle_F_semicircular_canal_I_utricular_macula_/425022", "title"=>"Comparison of vestibular morphology between <i>Slc26a4<sup>+/+</sup></i> mice (A, saccule; C, utricle; E, semicircular canal; G, utricular macula) and <i>Slc26a4<sup>tm1Dontuh/tm1Dontuh</sup></i> mice (B, saccule; D, utricle; F, semicircular canal; I, utricular macula).", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-07-21 01:23:42"}
  • {"files"=>["https://ndownloader.figshare.com/files/379373", "https://ndownloader.figshare.com/files/379411"], "description"=>"<div><p>Recessive mutations in the <em>SLC26A4</em> gene are a common cause of hereditary hearing impairment worldwide. Previous studies have demonstrated that different <em>SLC26A4</em> mutations may have different pathogenetic mechanisms. In the present study, we established a knock-in mouse model (i.e., <em>Slc26a4<sup>tm1Dontuh/tm1Dontuh</sup></em> mice) homozygous for the c.919-2A>G mutation, which is a common mutation in East Asians. Mice were then subjected to audiologic assessment, a battery of vestibular evaluations, and inner ear morphological studies. All <em>Slc26a4<sup>tm1Dontuh/tm1Dontuh</sup></em> mice revealed profound hearing loss, whereas 46% mice demonstrated pronounced head tilting and circling behaviors. There was a significant difference in the vestibular performance between wild-type and <em>Slc26a4<sup>tm1Dontuh/tm1Dontuh</sup></em> mice, especially those exhibiting circling behavior. Inner ear morphological examination of <em>Slc26a4<sup>tm1Dontuh/tm1Dontuh</sup></em> mice revealed an enlarged endolymphatic duct, vestibular aqueduct and sac, atrophy of stria vascularis, deformity of otoconia in the vestibular organs, consistent degeneration of cochlear hair cells, and variable degeneration of vestibular hair cells. Audiologic and inner ear morphological features of <em>Slc26a4<sup>tm1Dontuh/tm1Dontuh</sup></em> mice were reminiscent of those observed in humans. These features were also similar to those previously reported in both knock-out <em>Slc26a4<sup>−/−</sup></em> mice and <em>Slc26a4<sup>loop/loop</sup></em> mice with the <em>Slc26a4</em> p.S408F mutation, albeit the severity of vestibular hair cell degeneration appeared different among the three mouse strains.</p> </div>", "links"=>[], "tags"=>["knock-in", "mutation", "characterization", "pathology"], "article_id"=>134965, "categories"=>["Neuroscience", "Genetics"], "users"=>["Ying-Chang Lu", "Chen-Chi Wu", "Wen-Sheng Shen", "Ting-Hua Yang", "Te-Huei Yeh", "Pei-Jer Chen", "I-Shing Yu", "Shu-Wha Lin", "Jau-Min Wong", "Qing Chang", "Xi Lin", "Chuan-Jen Hsu"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0022150.s001", "https://dx.doi.org/10.1371/journal.pone.0022150.s002"], "stats"=>{"downloads"=>6, "page_views"=>17, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Establishment_of_a_Knock_In_Mouse_Model_with_the_SLC26A4_c_919_2A_G_Mutation_and_Characterization_of_Its_Pathology/134965", "title"=>"Establishment of a Knock-In Mouse Model with the <em>SLC26A4</em> c.919-2A>G Mutation and Characterization of Its Pathology", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2011-07-21 01:22:45"}
  • {"files"=>["https://ndownloader.figshare.com/files/753917"], "description"=>"<p>The <i>Slc26a4</i> transcripts from the allele with the c.919-2A>G mutation lead to a complete omission of exon 8 during the splicing process in <i>Slc26a4<sup>tm1Dontuh/tm1Dontuh</sup></i> mice.</p>", "links"=>[], "tags"=>["transcripts", "allele", "mutation", "omission", "exon", "splicing"], "article_id"=>424273, "categories"=>["Neuroscience", "Genetics"], "users"=>["Ying-Chang Lu", "Chen-Chi Wu", "Wen-Sheng Shen", "Ting-Hua Yang", "Te-Huei Yeh", "Pei-Jer Chen", "I-Shing Yu", "Shu-Wha Lin", "Jau-Min Wong", "Qing Chang", "Xi Lin", "Chuan-Jen Hsu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0022150.g001", "stats"=>{"downloads"=>4, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_Slc26a4_transcripts_from_the_allele_with_the_c_919_2A_G_mutation_lead_to_a_complete_omission_of_exon_8_during_the_splicing_process_in_Slc26a4_tm1Dontuh_tm1Dontuh_mice_/424273", "title"=>"The <i>Slc26a4</i> transcripts from the allele with the c.919-2A>G mutation lead to a complete omission of exon 8 during the splicing process in <i>Slc26a4<sup>tm1Dontuh/tm1Dontuh</sup></i> mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-07-21 01:11:13"}
  • {"files"=>["https://ndownloader.figshare.com/files/754099"], "description"=>"<p>All <i>Slc26a4<sup>tm1Dontuh/tm1Dontuh</sup></i> mice (n = 15) revealed profound hearing loss (>120 dB SPL) according to ABR at all frequencies, whereas all <i>Slc26a4<sup>+/tm1Dontuh</sup></i> mice (n = 15) and all <i>Slc26a4<sup>+/+</sup></i> mice (n = 15) revealed normal hearing.</p>", "links"=>[], "tags"=>["phenotypes"], "article_id"=>424465, "categories"=>["Neuroscience", "Genetics"], "users"=>["Ying-Chang Lu", "Chen-Chi Wu", "Wen-Sheng Shen", "Ting-Hua Yang", "Te-Huei Yeh", "Pei-Jer Chen", "I-Shing Yu", "Shu-Wha Lin", "Jau-Min Wong", "Qing Chang", "Xi Lin", "Chuan-Jen Hsu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0022150.g003", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Audiological_phenotypes_of_Slc26a4_tm1Dontuh_tm1Dontuh_mice_/424465", "title"=>"Audiological phenotypes of <i>Slc26a4<sup>tm1Dontuh/tm1Dontuh</sup></i> mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-07-21 01:14:25"}
  • {"files"=>["https://ndownloader.figshare.com/files/754525"], "description"=>"<p>The most significant change in stria vascularis in <i>Slc26a4<sup>tm1Dontuh/tm1Dontuh</sup></i> mice was diminished membrane invaginations of marginal cell with vacuolization changes. B, basal cell; I, intermediate cell; M, marginal cell; MI, membrane invaginations. Bar = 5 µm.</p>", "links"=>[], "tags"=>["electron", "micrographs", "stria", "vascularis", "mice"], "article_id"=>424879, "categories"=>["Neuroscience", "Genetics"], "users"=>["Ying-Chang Lu", "Chen-Chi Wu", "Wen-Sheng Shen", "Ting-Hua Yang", "Te-Huei Yeh", "Pei-Jer Chen", "I-Shing Yu", "Shu-Wha Lin", "Jau-Min Wong", "Qing Chang", "Xi Lin", "Chuan-Jen Hsu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0022150.g006", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Transmission_electron_micrographs_of_the_stria_vascularis_in_Slc26a4_mice_A_and_Slc26a4_tm1Dontuh_tm1Dontuh_mice_B_/424879", "title"=>"Transmission electron micrographs of the stria vascularis in <i>Slc26a4<sup>+/+</sup></i> mice (A) and <i>Slc26a4<sup>tm1Dontuh/tm1Dontuh</sup></i> mice (B).", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-07-21 01:21:19"}
  • {"files"=>["https://ndownloader.figshare.com/files/754337"], "description"=>"<p>Microscopic examination of the cochlear duct revealed severe endolymphatic hydrops with dilatation of scala media, degeneration of hair cells (B) and a significant atrophy of the stria vascularis (D) in <i>Slc26a4<sup>tm1Dontuh/tm1Dontuh</sup></i> mice. Degeneration of cochlear hair cells in <i>Slc26a4<sup>tm1Dontuh/tm1Dontuh</sup></i> mice was also revealed by fluorescence confocal microscopy (H). G, H: magnification of boxes G and H from figures E and F, respectively. B, bone; IHC, inner hair cells; OC, organ of Corti; OHC, outer hair cells; RM, Reissner's membrane; SL, spiral ligament; SV, stria vascularis; T, tunnel of Corti. Bar = 150 µm (A & B), 50 µm (C, D, E & F), and 10 µm (G & H).</p>", "links"=>[], "tags"=>["cochlear", "morphology", "mice"], "article_id"=>424701, "categories"=>["Neuroscience", "Genetics"], "users"=>["Ying-Chang Lu", "Chen-Chi Wu", "Wen-Sheng Shen", "Ting-Hua Yang", "Te-Huei Yeh", "Pei-Jer Chen", "I-Shing Yu", "Shu-Wha Lin", "Jau-Min Wong", "Qing Chang", "Xi Lin", "Chuan-Jen Hsu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0022150.g005", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Comparison_of_cochlear_morphology_between_Slc26a4_mice_A_C_E_G_and_Slc26a4_tm1Dontuh_tm1Dontuh_mice_B_D_F_H_/424701", "title"=>"Comparison of cochlear morphology between <i>Slc26a4<sup>+/+</sup></i> mice (A, C, E & G) and <i>Slc26a4<sup>tm1Dontuh/tm1Dontuh</sup></i> mice (B, D, F & H).", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-07-21 01:18:21"}
  • {"files"=>["https://ndownloader.figshare.com/files/754867"], "description"=>"<p>ND, not described.</p>*<p><i>Slc26a4<sup>loop/loop</sup></i>: mice homozygous for the p.S408F mutation.</p>", "links"=>[], "tags"=>["phenotypes", "strains", "segregating", "slc26a4"], "article_id"=>425230, "categories"=>["Neuroscience", "Genetics"], "users"=>["Ying-Chang Lu", "Chen-Chi Wu", "Wen-Sheng Shen", "Ting-Hua Yang", "Te-Huei Yeh", "Pei-Jer Chen", "I-Shing Yu", "Shu-Wha Lin", "Jau-Min Wong", "Qing Chang", "Xi Lin", "Chuan-Jen Hsu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0022150.t003", "stats"=>{"downloads"=>5, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Comparison_of_phenotypes_among_mouse_strains_segregating_different_slc26a4_mutations_/425230", "title"=>"Comparison of phenotypes among mouse strains segregating different slc26a4 mutations.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2011-07-21 01:27:10"}
  • {"files"=>["https://ndownloader.figshare.com/files/754905"], "description"=>"<p>*Observed at 3 weeks.</p>§<p>Tested at 8 weeks.</p>†<p><i>p</i>>0.05 as compared to wild type.</p>‡<p><i>p</i><0.01 as compared to wild type.</p>", "links"=>[], "tags"=>["vestibular", "genotypes", "circling"], "article_id"=>425266, "categories"=>["Neuroscience", "Genetics"], "users"=>["Ying-Chang Lu", "Chen-Chi Wu", "Wen-Sheng Shen", "Ting-Hua Yang", "Te-Huei Yeh", "Pei-Jer Chen", "I-Shing Yu", "Shu-Wha Lin", "Jau-Min Wong", "Qing Chang", "Xi Lin", "Chuan-Jen Hsu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0022150.t001", "stats"=>{"downloads"=>1, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Comparison_of_vestibular_features_according_to_the_genotypes_and_the_circling_behavior_/425266", "title"=>"Comparison of vestibular features according to the genotypes and the circling behavior.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2011-07-21 01:27:46"}
  • {"files"=>["https://ndownloader.figshare.com/files/754001"], "description"=>"<p>The expression of pendrin (stained in green) in spiral prominence and outer sulcus epithelial cells was not affected by the c.919-2A>G mutation. Degeneration of cochlear hair cells in <i>Slc26a4<sup>tm1Dontuh/tm1Dontuh</sup></i> mice was revealed by fluorescence confocal microscopy (D). (OC, organ of Corti; OS, outer sulcus epithelial cells; RC, root cells; SP, spiral prominence; SL, spiral ligament; SV, stria vascularis). Bar = 50 µm.</p>", "links"=>[], "tags"=>["pendrin", "mice"], "article_id"=>424362, "categories"=>["Neuroscience", "Genetics"], "users"=>["Ying-Chang Lu", "Chen-Chi Wu", "Wen-Sheng Shen", "Ting-Hua Yang", "Te-Huei Yeh", "Pei-Jer Chen", "I-Shing Yu", "Shu-Wha Lin", "Jau-Min Wong", "Qing Chang", "Xi Lin", "Chuan-Jen Hsu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0022150.g002", "stats"=>{"downloads"=>2, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Immunolocalization_of_pendrin_in_Slc26a4_mice_A_and_Slc26a4_tm1Dontuh_tm1Dontuh_mice_B_/424362", "title"=>"Immunolocalization of pendrin in <i>Slc26a4<sup>+/+</sup></i> mice (A) and <i>Slc26a4<sup>tm1Dontuh/tm1Dontuh</sup></i> mice (B).", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-07-21 01:12:42"}
  • {"files"=>["https://ndownloader.figshare.com/files/754203"], "description"=>"<p>The vestibular aqueduct and endolymphatic sac in the <i>Slc26a4<sup>tm1Dontuh/tm1Dontuh</sup></i> mice were enlarged (B & D) compared to those in the <i>Slc26a4<sup>+/+</sup></i> mice (A & C). (1, posterior semicircular canal; 2, superior semicircular canal; 3, subarcuate fossa; 4, fovea for endolymphatic sac (dot-dash line); 5, external orifice of vestibular aqueduct. Bar = 100 µm.</p>", "links"=>[], "tags"=>["aberrations", "vestibular", "aqueduct", "endolymphatic", "sac"], "article_id"=>424559, "categories"=>["Neuroscience", "Genetics"], "users"=>["Ying-Chang Lu", "Chen-Chi Wu", "Wen-Sheng Shen", "Ting-Hua Yang", "Te-Huei Yeh", "Pei-Jer Chen", "I-Shing Yu", "Shu-Wha Lin", "Jau-Min Wong", "Qing Chang", "Xi Lin", "Chuan-Jen Hsu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0022150.g004", "stats"=>{"downloads"=>2, "page_views"=>45, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Structural_aberrations_of_the_vestibular_aqueduct_and_endolymphatic_sac_in_mice_/424559", "title"=>"Structural aberrations of the vestibular aqueduct and endolymphatic sac in mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-07-21 01:15:59"}
  • {"files"=>["https://ndownloader.figshare.com/files/754816"], "description"=>"<p>*Fisher's exact test for categorical variables and Student's <i>t</i>-test for continuous variables.</p>†<p>Measured by calculating the number of areas of hair cell loss in 5 random sections pre mouse.</p>‡<p>Defined by decreased amount of otoconia in the saccule and utricle, formation of giant otoconia in the utricle and ectopic distribution of otoconia in the semicircular canal.</p>", "links"=>[], "tags"=>["vestibular", "morphological", "findings", "phenotype"], "article_id"=>425170, "categories"=>["Neuroscience", "Genetics"], "users"=>["Ying-Chang Lu", "Chen-Chi Wu", "Wen-Sheng Shen", "Ting-Hua Yang", "Te-Huei Yeh", "Pei-Jer Chen", "I-Shing Yu", "Shu-Wha Lin", "Jau-Min Wong", "Qing Chang", "Xi Lin", "Chuan-Jen Hsu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0022150.t002", "stats"=>{"downloads"=>1, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Comparison_of_vestibular_morphological_findings_according_to_the_vestibular_phenotype_of_circling_/425170", "title"=>"Comparison of vestibular morphological findings according to the vestibular phenotype of circling.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2011-07-21 01:26:10"}

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