Lipopolysaccharide-Induced Middle Ear Inflammation Disrupts the cochlear Intra-Strial Fluid–Blood Barrier through Down-Regulation of Tight Junction Proteins
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{"title"=>"Lipopolysaccharide-induced middle ear inflammation disrupts the cochlear intra-strial fluid-blood barrier through down-regulation of tight junction proteins", "type"=>"journal", "authors"=>[{"first_name"=>"Jinhui", "last_name"=>"Zhang", "scopus_author_id"=>"55776922200"}, {"first_name"=>"Songlin", "last_name"=>"Chen", "scopus_author_id"=>"56155346900"}, {"first_name"=>"Zhiqiang", "last_name"=>"Hou", "scopus_author_id"=>"56660738800"}, {"first_name"=>"Jing", "last_name"=>"Cai", "scopus_author_id"=>"57192989628"}, {"first_name"=>"Mingmin", "last_name"=>"Dong", "scopus_author_id"=>"7202127154"}, {"first_name"=>"Xiaorui", "last_name"=>"Shi", "scopus_author_id"=>"7402952928"}], "year"=>2015, "source"=>"PLoS ONE", "identifiers"=>{"isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "sgr"=>"84929493233", "pui"=>"604114771", "pmid"=>"25815897", "issn"=>"19326203", "scopus"=>"2-s2.0-84929493233", "doi"=>"10.1371/journal.pone.0122572"}, "id"=>"5328aa15-ad50-313e-b2ed-e87ff9439811", "abstract"=>"Middle ear infection (or inflammation) is the most common pathological condition that causes fluid to accumulate in the middle ear, disrupting cochlear homeostasis. Lipopolysaccharide, a product of bacteriolysis, activates macrophages and causes release of inflammatory cytokines. Many studies have shown that lipopolysaccharides cause functional and structural changes in the inner ear similar to that of inflammation. However, it is specifically not known how lipopolysaccharides affect the blood-labyrinth barrier in the stria vascularis (intra-strial fluid-blood barrier), nor what the underlying mechanisms are. In this study, we used a cell culture-based in vitro model and animal-based in vivo model, combined with immunohistochemistry and a vascular leakage assay, to investigate lipopolysaccharide effects on the integrity of the mouse intra-strial fluid-blood barrier. Our results show lipopolysaccharide-induced local infection significantly affects intra-strial fluid-blood barrier component cells. Pericytes and perivascular-resident macrophage-like melanocytes are particularly affected, and the morphological and functional changes in these cells are accompanied by substantial changes in barrier integrity. Significant vascular leakage is found in the lipopolysaccharide treated-animals. Consistent with the findings from the in vivo animal model, the permeability of the endothelial cell monolayer to FITC-albumin was significantly higher in the lipopolysaccharide-treated monolayer than in an untreated endothelial cell monolayer. Further study has shown the lipopolysaccharide-induced inflammation to have a major effect on the expression of tight junctions in the blood barrier. Lipopolysaccharide was also shown to cause high frequency hearing loss, corroborated by previous reports from other laboratories. Our findings show lipopolysaccharide-evoked middle ear infection disrupts inner ear fluid balance, and its particular effects on the intra-strial fluid-blood barrier, essential for cochlear homeostasis. The barrier is degraded as the expression of tight junction-associated proteins such as zona occludens 1, occludin, and vascular endothelial cadherin are down-regulated.", "link"=>"http://www.mendeley.com/research/lipopolysaccharideinduced-middle-ear-inflammation-disrupts-cochlear-intrastrial-fluidblood-barrier-t", "reader_count"=>9, "reader_count_by_academic_status"=>{"Researcher"=>4, "Other"=>3, "Student > Master"=>1, "Student > Bachelor"=>1}, "reader_count_by_user_role"=>{"Researcher"=>4, "Other"=>3, "Student > Master"=>1, "Student > Bachelor"=>1}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>1, "Medicine and Dentistry"=>3, "Agricultural and Biological Sciences"=>3, "Neuroscience"=>1, "Chemistry"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>3}, "Neuroscience"=>{"Neuroscience"=>1}, "Chemistry"=>{"Chemistry"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>3}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>1}}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1992551"], "description"=>"<p>LPS treatment increases vascular leakage. (<b>A</b>) Under normal conditions, the FITC-albumin is restricted to the vascular wall of the stria vascularis, and control animals show no sites of obvious leakage. <b>(B</b>) In LPS-treated animals, however, capillaries are permeable and multiple sites of vascular leakage are seen in the vessel of the stria vascularis. (<b>C</b>) Significantly more leakage of albumin-FITC is seen in LPS-treated animals than in control animals (**P<0.01). (<b>D</b>) Retention of FITC-albumin in isolated whole mounts of untreated cochlear stria vascularis is relatively low in non-LPS treated mice, and (<b>E</b>) retention in the LPS-treated mice is much stronger. (<b>F</b>) FITC-albumin fluorescence is significantly higher in the LPS-treated group than in control animals (**P<0.01). (<b>G</b>) and (<b>H</b>) The graph shows the effect of LPS on the permeability of tight junctions in an EC monolayer to FITC-dextran. Permeability was measured by determining the flux of FITC-dextran from the upper to the lower chamber of the Transwell filter. Data are expressed as means ± SD (**P<0.01).</p>", "links"=>[], "tags"=>["Vivo Animal Model", "Tight Junction Proteins Middle ear infection", "inflammation", "cochlear homeostasis", "barrier", "ear fluid balance", "cell monolayer", "lipopolysaccharide", "frequency hearing loss"], "article_id"=>1358880, "categories"=>["Biological Sciences"], "users"=>["Jinhui Zhang", "Songlin Chen", "Zhiqiang Hou", "Jing Cai", "Mingmin Dong", "Xiaorui Shi"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0122572.g003", "stats"=>{"downloads"=>0, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_LPS_treatment_increases_vascular_leakage_/1358880", "title"=>"LPS treatment increases vascular leakage.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-03-27 03:54:43"}
  • {"files"=>["https://ndownloader.figshare.com/files/1992549"], "description"=>"<p>Confocal maximum projection images of the tympanic membrane and mucosa of the middle ear in control (A, B and C) and LPS-treated mice (D, E and F). (A) is a low magnification image showing the normal tympanic membrane (TM) and mucosa in the middle ear (mucosa/ME) of a control animal. (<b>B</b>) and (<b>C</b>) are high magnification images of the TM and mucosa/ME in a control animal. (<b>D</b>) is a low magnification image showing thickening of the TM and swelling of the mucosa/ME typically observed in LPS-treated animals. (<b>E</b>) and (<b>F</b>) are high magnification images showing a large number of infiltrated cells expressing F4/80 (red, arrow) and positive for GS-IB4 (green, arrow) in the region of the TM and mucosa/SEM in LPS-treated animals. TM = tympanic membrane; ME = middle ear.</p>", "links"=>[], "tags"=>["Vivo Animal Model", "Tight Junction Proteins Middle ear infection", "inflammation", "cochlear homeostasis", "barrier", "ear fluid balance", "cell monolayer", "lipopolysaccharide", "frequency hearing loss"], "article_id"=>1358878, "categories"=>["Biological Sciences"], "users"=>["Jinhui Zhang", "Songlin Chen", "Zhiqiang Hou", "Jing Cai", "Mingmin Dong", "Xiaorui Shi"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0122572.g001", "stats"=>{"downloads"=>0, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Confocal_projection_images_of_the_tympanic_membrane_and_mucosa_of_the_middle_ear_in_control_and_LPS_treated_mice_/1358878", "title"=>"Confocal projection images of the tympanic membrane and mucosa of the middle ear in control and LPS-treated mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-03-27 03:54:43"}
  • {"files"=>["https://ndownloader.figshare.com/files/1992562"], "description"=>"<p>LPS damages hearing function. (<b>A</b>) and (<b>B</b>) show the mean value of hearing threshold levels measured by ABR in control and LPS-treated groups. The ABR threshold in LPS-treated animals is significantly elevated at 4 kHz, 16 kHz, 24 kHz, and 32 kHz (**P<0.01).</p>", "links"=>[], "tags"=>["Vivo Animal Model", "Tight Junction Proteins Middle ear infection", "inflammation", "cochlear homeostasis", "barrier", "ear fluid balance", "cell monolayer", "lipopolysaccharide", "frequency hearing loss"], "article_id"=>1358883, "categories"=>["Biological Sciences"], "users"=>["Jinhui Zhang", "Songlin Chen", "Zhiqiang Hou", "Jing Cai", "Mingmin Dong", "Xiaorui Shi"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0122572.g005", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_LPS_causes_hearing_loss_/1358883", "title"=>"LPS causes hearing loss.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-03-27 03:54:43"}
  • {"files"=>["https://ndownloader.figshare.com/files/1992561"], "description"=>"<p>LPS affects the expression of tight-junction proteins. (<b>A</b>) qRT-PCR shows mRNA expression for ZO-1, occludin, and ve-cadherin in control and LPS treated animals (**P<0.01, *P <0.05). (<b>B</b>) Quantitative analysis of ZO-1, occludin, and ve-cadherin protein expression in control and LPS-treated groups (*P<0.05). (<b>C</b>) Western blot analysis shows significantly decreased expression of ZO-1, occludin, and ve-cadherin in the LPS-treated group. (<b>D</b>) TEM images show TJ contact points are extensive in control mice (Left, zoomed inset). The left window (zoomed inset) displays a higher magnification of the TJs between ECs. (<b>E</b>) Alterations in the TJ ultrastructure are seen in the LPS-treated mice. TJ contact points were reduced in number in response to LPS (left, inset). (<b>F</b>) qRT-PCR shows mRNA for ZO-1, occludin, and ve-cadherin in the endothelial monolayer of the control and LPS group at different time points (**P<0.01, *P<0.05). (<b>G</b>-<b>I</b>) Quantitative analysis of protein expression for ZO-1, occludin, and ve-cadherin in control and LPS-treated groups (**P<0.01). (<b>J</b>-<b>L</b>) In-cell Western blot analysis of tight junction protein expression. (<b>M</b>-<b>R</b>) Representative confocal maximum projection images show immunofluorescent labeling of tight junction protein expression, including ZO-1, occludin, and ve-cadherin, in the EC monolayer.</p>", "links"=>[], "tags"=>["Vivo Animal Model", "Tight Junction Proteins Middle ear infection", "inflammation", "cochlear homeostasis", "barrier", "ear fluid balance", "cell monolayer", "lipopolysaccharide", "frequency hearing loss"], "article_id"=>1358882, "categories"=>["Biological Sciences"], "users"=>["Jinhui Zhang", "Songlin Chen", "Zhiqiang Hou", "Jing Cai", "Mingmin Dong", "Xiaorui Shi"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0122572.g004", "stats"=>{"downloads"=>5, "page_views"=>130, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_LPS_affects_the_expression_of_tight_junction_proteins_/1358882", "title"=>"LPS affects the expression of tight-junction proteins.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-03-27 03:54:43"}
  • {"files"=>["https://ndownloader.figshare.com/files/1992563"], "description"=>"<p>Antibodies applied.</p>", "links"=>[], "tags"=>["Vivo Animal Model", "Tight Junction Proteins Middle ear infection", "inflammation", "cochlear homeostasis", "barrier", "ear fluid balance", "cell monolayer", "lipopolysaccharide", "frequency hearing loss"], "article_id"=>1358884, "categories"=>["Biological Sciences"], "users"=>["Jinhui Zhang", "Songlin Chen", "Zhiqiang Hou", "Jing Cai", "Mingmin Dong", "Xiaorui Shi"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0122572.t001", "stats"=>{"downloads"=>7, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Antibodies_applied_/1358884", "title"=>"Antibodies applied.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2015-03-27 03:54:43"}
  • {"files"=>["https://ndownloader.figshare.com/files/1992550"], "description"=>"<p>LPS treatment “activates” PCs and PVM/Ms in the intra-strial fluid–blood barrier. (<b>A</b>) and (<b>B</b>) Cochlear cross-sections show slight hydrops in LPS-treated animals (black arrows/stars). (<b>C</b>) A representative confocal 3D maximum projection shows normal, inactivated PCs <i>in vivo</i> (in transgenic mice with fluorescent labeled NG2, red) to have a flat and slender cell body. The cell morphology is clearly shown in the enlarged image from region of (a) in <b><a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0122572#pone.0122572.g002\" target=\"_blank\">Fig 2C</a></b>. (<b>D</b>) LPS stimulated PCs display a “prominent round” cell body in less physical contact with the strial capillary and release a large number of particles which can be easily seen in the enlarged image from region of (b) in <b><a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0122572#pone.0122572.g002\" target=\"_blank\">Fig 2D</a></b> (indicated by the arrow). (<b>E</b>) PVM/M distribution on strial capillaries, shown labeled with GS-IB4 in a control animal. <b>Fig E</b> (c) is an enlarged image from region of (c) in <b><a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0122572#pone.0122572.g002\" target=\"_blank\">Fig 2E</a>.</b> The PVM/M enshrouds a capillary network with its slander dendritic branches (indicated by the arrow). (<b>F</b>) PVM/Ms in LPS-treated animals display a reduced amount of branching, as well as withdrawal of ramifications. PVM/Ms in LPS-treated animals are in less physical contact with capillaries. A subset of the PVM/Ms is activated, indicated by display of a terminal galactopyranosyl group on the membrane surface and binding with GS-IB4. <b><a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0122572#pone.0122572.g002\" target=\"_blank\">Fig 2F</a></b> (d) is an enlarged image from region of (c) in <b><a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0122572#pone.0122572.g002\" target=\"_blank\">Fig 2F</a>.</b> Here the PVM/M is shown still surrounding the capillary network, but with withdrawal of dendritic branches (arrow). <b>(G)</b> Primary cell lines of cultured PCs are large, flat, stellate shaped cells, with a broadfilopod morphology. (<b>H)</b> LPS treatment for 48 hours causes PCs to branch and release a large number of particles. (<b>I</b>) Primary cell lines of cultured PVM/Ms under control conditions display a unique pattern of dendritic processes. GS-IB4 fluorescence in the control (non-LPS treated) cell line was undetectable. (<b>J</b>) Some perivascular resident macrophages are activated, displaying a terminal galactopyranosyl group on the membrane surface and binding the lectin Griffonia simplicifolia-IB4 (arrows, GS-IB4). The data show PCs and PVM/Ms are highly responsive to LPS. The control and LPS-treated groups show clear signs of morphological changes.</p>", "links"=>[], "tags"=>["Vivo Animal Model", "Tight Junction Proteins Middle ear infection", "inflammation", "cochlear homeostasis", "barrier", "ear fluid balance", "cell monolayer", "lipopolysaccharide", "frequency hearing loss"], "article_id"=>1358879, "categories"=>["Biological Sciences"], "users"=>["Jinhui Zhang", "Songlin Chen", "Zhiqiang Hou", "Jing Cai", "Mingmin Dong", "Xiaorui Shi"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0122572.g002", "stats"=>{"downloads"=>2, "page_views"=>17, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_LPS_treatment_8220_activates_8221_PCs_and_PVM_Ms_in_the_intra_strial_fluid_8211_blood_barrier_/1358879", "title"=>"LPS treatment “activates” PCs and PVM/Ms in the intra-strial fluid–blood barrier.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-03-27 03:54:43"}

PMC Usage Stats | Further Information

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Relative Metric

{"start_date"=>"2015-01-01T00:00:00Z", "end_date"=>"2015-12-31T00:00:00Z", "subject_areas"=>[]}
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