Effect of P2X7 Receptor Knockout on AQP-5 Expression of Type I Alveolar Epithelial Cells
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{"title"=>"Effect of P2X7 receptor knockout on AQP-5 expression of type I alveolar epithelial cells", "type"=>"journal", "authors"=>[{"first_name"=>"Georg", "last_name"=>"Ebeling", "scopus_author_id"=>"54879284800"}, {"first_name"=>"Robert", "last_name"=>"Bläsche", "scopus_author_id"=>"8389051400"}, {"first_name"=>"Falk", "last_name"=>"Hofmann", "scopus_author_id"=>"56230102700"}, {"first_name"=>"Antje", "last_name"=>"Augstein", "scopus_author_id"=>"15118855000"}, {"first_name"=>"Michael", "last_name"=>"Kasper", "scopus_author_id"=>"7102938719"}, {"first_name"=>"Kathrin", "last_name"=>"Barth", "scopus_author_id"=>"7102952539"}], "year"=>2014, "source"=>"PLoS ONE", "identifiers"=>{"sgr"=>"84903271932", "pui"=>"373396627", "scopus"=>"2-s2.0-84903271932", "issn"=>"19326203", "pmid"=>"24941004", "doi"=>"10.1371/journal.pone.0100282"}, "id"=>"d14d450c-c775-3821-86f1-2107a404483f", "abstract"=>"P2X7 receptors, ATP-gated cation channels, are specifically expressed in alveolar epithelial cells. The pathophysiological function of this lung cell type, except a recently reported putative involvement in surfactant secretion, is unknown. In addition, P2X7 receptor-deficient mice show reduced inflammation and lung fibrosis after exposure with bleomycin. To elucidate the role of the P2X7 receptor in alveolar epithelial type I cells we characterized the pulmonary phenotype of P2X7 receptor knockout mice by using immunohistochemistry, western blot analysis and real-time RT PCR. No pathomorphological signs of fibrosis were found. Results revealed, however, a remarkable loss of aquaporin-5 protein and mRNA in young knockout animals. Additional in vitro experiments with bleomycin treated precision cut lung slices showed a greater sensitivity of the P2X7 receptor knockout mice in terms of aquaporin-5 reduction as wild type animals. Finally, P2X7 receptor function was examined by using the alveolar epithelial cell lines E10 and MLE-12 for stimulation experiments with bleomycin. The in vitro activation of P2X7 receptor was connected with an increase of aquaporin-5, whereas the inhibition of the receptor with oxidized ATP resulted in down regulation of aquaporin-5. The early loss of aquaporin-5 which can be found in different pulmonary fibrosis models does not implicate a specific pathogenetic role during fibrogenesis.", "link"=>"http://www.mendeley.com/research/effect-p2x7-receptor-knockout-aqp5-expression-type-i-alveolar-epithelial-cells", "reader_count"=>15, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Researcher"=>5, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>4, "Student > Postgraduate"=>1, "Student > Master"=>2, "Other"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Researcher"=>5, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>4, "Student > Postgraduate"=>1, "Student > Master"=>2, "Other"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>1, "Medicine and Dentistry"=>3, "Agricultural and Biological Sciences"=>5, "Pharmacology, Toxicology and Pharmaceutical Science"=>1, "Chemistry"=>2, "Immunology and Microbiology"=>3}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>3}, "Chemistry"=>{"Chemistry"=>2}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>3}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>5}, "Unspecified"=>{"Unspecified"=>1}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1538591"], "description"=>"<p>Primers used for quantitative real-time RT PCR.</p>", "links"=>[], "tags"=>["anatomy", "biological tissue", "epithelium", "Epithelial cells", "histology", "Respiratory system", "cell biology", "Cellular types", "Molecular cell biology", "immunology", "Clinical immunology", "Pulmonary immunology", "Histochemistry and cytochemistry techniques", "Immunohistochemistry techniques", "Immunohistochemical analysis", "Immunologic techniques", "Model organisms", "Animal models", "Mouse models", "quantitative", "Real-time", "rt"], "article_id"=>1060602, "categories"=>["Biological Sciences"], "users"=>["Georg Ebeling", "Robert Bläsche", "Falk Hofmann", "Antje Augstein", "Michael Kasper", "Kathrin Barth"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0100282.t002", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Primers_used_for_quantitative_real_time_RT_PCR_/1060602", "title"=>"Primers used for quantitative real-time RT PCR.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2014-06-18 02:51:28"}
  • {"files"=>["https://ndownloader.figshare.com/files/1538586"], "description"=>"<p>mRNA expression, analysis of corresponding protein content and immunoperoxidase demonstration of Cav-1 (A), T1α (B) and Cx43 (C) in lungs from wild type and <i>P2rx7<sup>−/−</sup></i> mice aged 2–3 months compared to the particular protein content in wild type and <i>P2rx7<sup>−/−</sup></i> mice aged 11–12 months. Quantitative real-time RT PCR was done by using <i>Rpl32</i> and <i>Polr2a</i> as housekeeping genes. Results are shown median with whiskers extending to the minimum and maximum (n = 18). Equal protein amounts were used in SDS-PAGE and analyzed by western blot with antibodies against the particular protein. Protein levels are normalized to γ-Tub and are shown median with whiskers extending to the minimum and maximum (n<sub>(2–3 months)</sub> = 15, n<sub>(11–12 months)</sub> = 10) and one representative blot is pictured. Scale bar correlates to 50 µm in immunoperoxidase demonstrations.</p>", "links"=>[], "tags"=>["anatomy", "biological tissue", "epithelium", "Epithelial cells", "histology", "Respiratory system", "cell biology", "Cellular types", "Molecular cell biology", "immunology", "Clinical immunology", "Pulmonary immunology", "Histochemistry and cytochemistry techniques", "Immunohistochemistry techniques", "Immunohistochemical analysis", "Immunologic techniques", "Model organisms", "Animal models", "Mouse models", "cx43", "mice"], "article_id"=>1060597, "categories"=>["Biological Sciences"], "users"=>["Georg Ebeling", "Robert Bläsche", "Falk Hofmann", "Antje Augstein", "Michael Kasper", "Kathrin Barth"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0100282.g003", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Protein_content_and_expression_of_Cav_1_T1_and_Cx43_in_P2rx7_8722_8722_mice_lungs_/1060597", "title"=>"Protein content and expression of Cav-1, T1α and Cx43 in <i>P2rx7<sup>−/−</sup></i> mice lungs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-06-18 02:51:28"}
  • {"files"=>["https://ndownloader.figshare.com/files/1538585"], "description"=>"<p>Immunoperoxidase demonstration of Sirius Red (A), CD44v10 (B) and CC3 (C) in lungs from two cohorts of wild type and <i>P2rx7<sup>−/−</sup></i> mice (younger age bracket 2–3 months; older age bracket 11–12 months). Scale bar 50 µm.</p>", "links"=>[], "tags"=>["anatomy", "biological tissue", "epithelium", "Epithelial cells", "histology", "Respiratory system", "cell biology", "Cellular types", "Molecular cell biology", "immunology", "Clinical immunology", "Pulmonary immunology", "Histochemistry and cytochemistry techniques", "Immunohistochemistry techniques", "Immunohistochemical analysis", "Immunologic techniques", "Model organisms", "Animal models", "Mouse models", "staining", "immunohistochemical", "cd44v10", "cc3", "mice"], "article_id"=>1060596, "categories"=>["Biological Sciences"], "users"=>["Georg Ebeling", "Robert Bläsche", "Falk Hofmann", "Antje Augstein", "Michael Kasper", "Kathrin Barth"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0100282.g002", "stats"=>{"downloads"=>1, "page_views"=>48, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Sirius_Red_staining_and_immunohistochemical_staining_of_CD44v10_and_CC3_in_P2rx7_8722_8722_mice_lungs_/1060596", "title"=>"Sirius Red staining and immunohistochemical staining of CD44v10 and CC3 in <i>P2rx7<sup>−/−</sup></i> mice lungs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-06-18 02:51:28"}
  • {"files"=>["https://ndownloader.figshare.com/files/1538590"], "description"=>"<p>Primary Antibodies used for Western Blot (WB) and Immunohistochemistry (IHC).</p>", "links"=>[], "tags"=>["anatomy", "biological tissue", "epithelium", "Epithelial cells", "histology", "Respiratory system", "cell biology", "Cellular types", "Molecular cell biology", "immunology", "Clinical immunology", "Pulmonary immunology", "Histochemistry and cytochemistry techniques", "Immunohistochemistry techniques", "Immunohistochemical analysis", "Immunologic techniques", "Model organisms", "Animal models", "Mouse models", "antibodies", "blot", "immunohistochemistry"], "article_id"=>1060601, "categories"=>["Biological Sciences"], "users"=>["Georg Ebeling", "Robert Bläsche", "Falk Hofmann", "Antje Augstein", "Michael Kasper", "Kathrin Barth"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0100282.t001", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Primary_Antibodies_used_for_Western_Blot_WB_and_Immunohistochemistry_IHC_/1060601", "title"=>"Primary Antibodies used for Western Blot (WB) and Immunohistochemistry (IHC).", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2014-06-18 02:51:28"}
  • {"files"=>["https://ndownloader.figshare.com/files/1538588"], "description"=>"<p>Analysis of protein content in MLE-12 cells (A) and E10 cells (B) after BLM treatment: Cells were treated with 100 mU/ml BLM, 100 µM oxATP with BLM (100 mU/ml) and 100 µM oxATP for 24 h. oxATP was added 2 h prior to BLM treatment. Untreated cells were used as control. For SDS-Page equal protein amounts of cell lysates were used and analyzed by western blot with antibodies against P2X7R and AQP-5. Protein levels are normalized to γ-Tub and are shown mean ± SEM (n = 6) in relation to control. One representative blot is pictured (* p<0.05).</p>", "links"=>[], "tags"=>["anatomy", "biological tissue", "epithelium", "Epithelial cells", "histology", "Respiratory system", "cell biology", "Cellular types", "Molecular cell biology", "immunology", "Clinical immunology", "Pulmonary immunology", "Histochemistry and cytochemistry techniques", "Immunohistochemistry techniques", "Immunohistochemical analysis", "Immunologic techniques", "Model organisms", "Animal models", "Mouse models", "aqp-5", "p2x7r"], "article_id"=>1060599, "categories"=>["Biological Sciences"], "users"=>["Georg Ebeling", "Robert Bläsche", "Falk Hofmann", "Antje Augstein", "Michael Kasper", "Kathrin Barth"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0100282.g005", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Dependency_of_AQP_5_on_P2X7R_expression_/1060599", "title"=>"Dependency of AQP-5 on P2X7R expression.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-06-18 02:51:28"}
  • {"files"=>["https://ndownloader.figshare.com/files/1538589"], "description"=>"<p>Analysis of protein content in MLE-12 cells (A) and E10 cells (B) after BLM treatment: Cells were treated with 100 mU/ml BLM for 24 h, 48 h and 72 h. Untreated cells after 24 h, 48 h and 72 h were used as control. For SDS-Page equal protein amounts of cell lysates were used and analyzed by western blot with antibodies against P2X7R and AQP-5. Protein levels are normalized to γ-Tub and are shown mean ± SEM (n = 3) in relation to control. One representative blot is pictured (* p<0.05).</p>", "links"=>[], "tags"=>["anatomy", "biological tissue", "epithelium", "Epithelial cells", "histology", "Respiratory system", "cell biology", "Cellular types", "Molecular cell biology", "immunology", "Clinical immunology", "Pulmonary immunology", "Histochemistry and cytochemistry techniques", "Immunohistochemistry techniques", "Immunohistochemical analysis", "Immunologic techniques", "Model organisms", "Animal models", "Mouse models", "p2x7r"], "article_id"=>1060600, "categories"=>["Biological Sciences"], "users"=>["Georg Ebeling", "Robert Bläsche", "Falk Hofmann", "Antje Augstein", "Michael Kasper", "Kathrin Barth"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0100282.g006", "stats"=>{"downloads"=>0, "page_views"=>15, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_AQP_5_and_P2X7R_expression_over_the_course_of_time_/1060600", "title"=>"AQP-5 and P2X7R expression over the course of time.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-06-18 02:51:28"}
  • {"files"=>["https://ndownloader.figshare.com/files/1538587"], "description"=>"<p>Immunoperoxidase demonstration of AQP-5 (A) and Sirius Red staining (B) of wild type and <i>P2rx7<sup>−/−</sup></i> mice lungs after BLM treatment with 200 mU/ml for 72 h in PCLS. Untreated lung slices after 72 h were used as controls. Scale bar 50 µm. For analysis of AQP-5 protein content, equal protein amounts were used in SDS-PAGE and analyzed by western blot with the antibody against the AQP-5 (C). BLM treated and untreated PCLS from wild type and <i>P2rx7<sup>−/−</sup></i> mice lungs were compared statistically. Relative protein levels AQP-5/γ-Tub are shown mean ± SEM (n = 4) and one representative blot is pictured (* p<0.05).</p>", "links"=>[], "tags"=>["anatomy", "biological tissue", "epithelium", "Epithelial cells", "histology", "Respiratory system", "cell biology", "Cellular types", "Molecular cell biology", "immunology", "Clinical immunology", "Pulmonary immunology", "Histochemistry and cytochemistry techniques", "Immunohistochemistry techniques", "Immunohistochemical analysis", "Immunologic techniques", "Model organisms", "Animal models", "Mouse models", "aqp-5", "mice", "compared", "knockout", "blm"], "article_id"=>1060598, "categories"=>["Biological Sciences"], "users"=>["Georg Ebeling", "Robert Bläsche", "Falk Hofmann", "Antje Augstein", "Michael Kasper", "Kathrin Barth"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0100282.g004", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Collagen_and_AQP_5_protein_content_in_wild_type_mice_compared_to_P2rx7_8722_8722_knockout_mice_after_BLM_treatment_/1060598", "title"=>"Collagen and AQP-5 protein content in wild type mice compared to <i>P2rx7<sup>−/−</sup></i> knockout mice after BLM treatment.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-06-18 02:51:28"}
  • {"files"=>["https://ndownloader.figshare.com/files/1538584"], "description"=>"<p>Immunohistochemical staining of AQP-5, <i>Aqp5</i> mRNA expression and analysis of AQP-5 protein content in lungs from wild type and <i>P2rx7<sup>−/−</sup></i> mice aged (A) 2–3 months and (B) 11–12 months. Scale bar correlates to 50 µm in immunoperoxidase demonstrations. Quantitative real-time RT PCR was done by using <i>Rpl32</i> and <i>Polr2a</i> as housekeeping genes. Results are shown median with whiskers extending to the minimum and maximum (n<sub>(2–3 months)</sub> = 18, n<sub>(11–12 months)</sub> = 9). Equal protein amounts were used in SDS-PAGE and analyzed by western blot with the antibody against AQP-5. Relative protein levels AQP-5/γ-Tub are shown median with whiskers extending to the minimum and maximum (n<sub>(2–3 months)</sub> = 15, n<sub>(11–12 months)</sub> = 15) and one representative blot is pictured (* p<0.05).</p>", "links"=>[], "tags"=>["anatomy", "biological tissue", "epithelium", "Epithelial cells", "histology", "Respiratory system", "cell biology", "Cellular types", "Molecular cell biology", "immunology", "Clinical immunology", "Pulmonary immunology", "Histochemistry and cytochemistry techniques", "Immunohistochemistry techniques", "Immunohistochemical analysis", "Immunologic techniques", "Model organisms", "Animal models", "Mouse models", "aqp-5", "mice"], "article_id"=>1060595, "categories"=>["Biological Sciences"], "users"=>["Georg Ebeling", "Robert Bläsche", "Falk Hofmann", "Antje Augstein", "Michael Kasper", "Kathrin Barth"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0100282.g001", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Protein_content_and_expression_of_AQP_5_in_P2rx7_8722_8722_mice_lungs_/1060595", "title"=>"Protein content and expression of AQP-5 in <i>P2rx7<sup>−/−</sup></i> mice lungs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-06-18 02:51:28"}

PMC Usage Stats | Further Information

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Relative Metric

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