Cdc42-Dependent Activation of NADPH Oxidase Is Involved in Ethanol-Induced Neuronal Oxidative Stress
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{"title"=>"Cdc42-dependent activation of NADPH oxidase is involved in Ethanol-Induced neuronal oxidative stress", "type"=>"journal", "authors"=>[{"first_name"=>"Xin", "last_name"=>"Wang", "scopus_author_id"=>"56336437600"}, {"first_name"=>"Zunji", "last_name"=>"Ke", "scopus_author_id"=>"8637113400"}, {"first_name"=>"Gang", "last_name"=>"Chen", "scopus_author_id"=>"57032029400"}, {"first_name"=>"Mei", "last_name"=>"Xu", "scopus_author_id"=>"35338672600"}, {"first_name"=>"Kimberly A.", "last_name"=>"Bower", "scopus_author_id"=>"8776778400"}, {"first_name"=>"Jacqueline A.", "last_name"=>"Frank", "scopus_author_id"=>"36631774100"}, {"first_name"=>"Zhuo", "last_name"=>"Zhang", "scopus_author_id"=>"8758845800"}, {"first_name"=>"Xianglin", "last_name"=>"Shi", "scopus_author_id"=>"7402953332"}, {"first_name"=>"Jia", "last_name"=>"Luo", "scopus_author_id"=>"25626675700"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"scopus"=>"2-s2.0-84861468560", "sgr"=>"84861468560", "issn"=>"19326203", "doi"=>"10.1371/journal.pone.0038075", "pmid"=>"22662267", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "pui"=>"364883018"}, "id"=>"2b302efe-4b1a-388d-a287-76494e8b9010", "abstract"=>"It has been suggested that excessive reactive oxygen species (ROS) and oxidative stress play an important role in ethanol-induced damage to both the developing and mature central nervous system (CNS). The mechanisms underlying ethanol-induced neuronal ROS, however, remain unclear. In this study, we investigated the role of NADPH oxidase (NOX) in ethanol-induced ROS generation. We demonstrated that ethanol activated NOX and inhibition of NOX reduced ethanol-promoted ROS generation. Ethanol significantly increased the expression of p47(phox) and p67(phox), the essential subunits for NOX activation in cultured neuronal cells and the cerebral cortex of infant mice. Ethanol caused serine phosphorylation and membrane translocation of p47(phox) and p67(phox), which were prerequisites for NOX assembly and activation. Knocking down p47(phox) with the small interfering RNA was sufficient to attenuate ethanol-induced ROS production and ameliorate ethanol-mediated oxidative damage, which is indicated by a decrease in protein oxidation and lipid peroxidation. Ethanol activated cell division cycle 42 (Cdc42) and overexpression of a dominant negative (DN) Cdc42 abrogate ethanol-induced NOX activation and ROS generation. These results suggest that Cdc42-dependent NOX activation mediates ethanol-induced oxidative damages to neurons.", "link"=>"http://www.mendeley.com/research/cdc42dependent-activation-nadph-oxidase-involved-ethanolinduced-neuronal-oxidative-stress", "reader_count"=>19, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>2, "Student > Doctoral Student"=>2, "Researcher"=>4, "Student > Ph. D. Student"=>5, "Student > Master"=>1, "Student > Bachelor"=>2, "Professor"=>1, "Other"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>2, "Student > Doctoral Student"=>2, "Researcher"=>4, "Student > Ph. D. Student"=>5, "Student > Master"=>1, "Student > Bachelor"=>2, "Professor"=>1, "Other"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>3, "Biochemistry, Genetics and Molecular Biology"=>3, "Nursing and Health Professions"=>1, "Agricultural and Biological Sciences"=>8, "Medicine and Dentistry"=>2, "Neuroscience"=>2}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>2}, "Neuroscience"=>{"Neuroscience"=>2}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>8}, "Nursing and Health Professions"=>{"Nursing and Health Professions"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}, "Unspecified"=>{"Unspecified"=>3}}, "reader_count_by_country"=>{"Brazil"=>1, "Portugal"=>1}, "group_count"=>1}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/633150"], "description"=>"<p>SH-SY5Y cells were transfected with p47<sup>phox</sup> siRNA for 48 hours as described under the <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0038075#s2\" target=\"_blank\">Materials and Methods</a>. The expression of p47<sup>phox</sup> was determined by immunoblotting (top panel). SH-SY5Y cells were transfected with p47<sup>phox</sup> siRNA for 48 hours and exposed to ethanol (0 or 0.4%) for 24 hours. The relative amount of ROS was measured as described above (bottom panel). The data are expressed as the mean ± SEM of three independent experiments. * denotes statistically significant difference from control cells (<i>p</i><0.05); # denotes statistically significant difference from ethanol-treated cells (<i>p</i><0.05).</p>", "links"=>[], "tags"=>["sirna", "ethanol-induced", "ros"], "article_id"=>303632, "categories"=>["Biochemistry", "Neuroscience", "Cell Biology", "Mental Health"], "users"=>["Xin Wang", "Zunji Ke", "Gang Chen", "Mei Xu", "Kimberly A. Bower", "Jacqueline A. Frank", "Zhuo Zhang", "Xianglin Shi", "Jia Luo"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0038075.g003", "stats"=>{"downloads"=>1, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_p47_phox_siRNA_on_ethanol_induced_ROS_generation_/303632", "title"=>"Effect of p47<sup>phox</sup> siRNA on ethanol-induced ROS generation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 04:43:28"}
  • {"files"=>["https://ndownloader.figshare.com/files/632939"], "description"=>"<p><b>A</b>. SH-SY5Y cells were exposed to ethanol (0 or 0.4%) for indicated times. NOX activity was determined by lucigenin chemiluminescence assay as described under the <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0038075#s2\" target=\"_blank\">Materials and Methods</a>. <b>B</b>. SH-SY5Y cells were pretreated with apocynin (0 or 50 μM) for 2 hours followed by exposure to 0.4% ethanol for 24 hours. Cells were labeled with DCFDA (10 μM) as described under the <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0038075#s2\" target=\"_blank\">Materials and Methods</a> and relative intensity of DCFDA (cellular ROS concentration) was measured and presented. The data are expressed as the mean ± SEM of three independent experiments. * denotes statistically significant difference from control cells (<i>p</i><0.05); # denotes statistically significant difference from ethanol-treated cells (<i>p</i><0.05).</p>", "links"=>[], "tags"=>["ethanol", "nox", "activation", "ros"], "article_id"=>303421, "categories"=>["Biochemistry", "Neuroscience", "Cell Biology", "Mental Health"], "users"=>["Xin Wang", "Zunji Ke", "Gang Chen", "Mei Xu", "Kimberly A. Bower", "Jacqueline A. Frank", "Zhuo Zhang", "Xianglin Shi", "Jia Luo"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0038075.g001", "stats"=>{"downloads"=>12, "page_views"=>20, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_ethanol_on_NOX_activation_and_ROS_generation_/303421", "title"=>"Effect of ethanol on NOX activation and ROS generation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 04:42:14"}
  • {"files"=>["https://ndownloader.figshare.com/files/633258"], "description"=>"<p>SH-SY5Y cells were transfected with p47<sup>phox</sup> siRNA for 48 hours and treated with ethanol (0 or 0.4%) for 48 hours. Protein oxidation and lipid peroxidation were determined by AOPP and MDA assay, respectively as described under the <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0038075#s2\" target=\"_blank\">Materials and Methods</a>. The data are expressed as the mean ± SEM of three independent experiments. * denotes statistically significant difference from control cells (<i>p</i><0.05); # denotes statistically significant difference from ethanol-treated cells (<i>p</i><0.05).</p>", "links"=>[], "tags"=>["sirna", "ethanol-induced", "oxidative"], "article_id"=>303738, "categories"=>["Biochemistry", "Neuroscience", "Cell Biology", "Mental Health"], "users"=>["Xin Wang", "Zunji Ke", "Gang Chen", "Mei Xu", "Kimberly A. Bower", "Jacqueline A. Frank", "Zhuo Zhang", "Xianglin Shi", "Jia Luo"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0038075.g004", "stats"=>{"downloads"=>1, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_p47_phox_siRNA_on_ethanol_induced_oxidative_damage_/303738", "title"=>"Effect of p47<sup>phox</sup> siRNA on ethanol-induced oxidative damage.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 04:44:02"}
  • {"files"=>["https://ndownloader.figshare.com/files/633060"], "description"=>"<p><b>A</b>. Top: SH-SY5Y cells were treated with ethanol (0 or 0.4%) for specified times. The expression of p47<sup>phox</sup> and p67<sup>phox</sup> was determined by immunoblotting. The expression of actin served as an internal loading control. Relative amounts of p47<sup>phox</sup> and p67<sup>phox</sup> was measured by densitometry and normalized to the expression of actin. Bottom: 7-days-old mice were exposed to ethanol as described under the <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0038075#s2\" target=\"_blank\">Materials and Methods</a>. The expression of p47<sup>phox</sup> and p67<sup>phox</sup> in the prefrontal cortex was determined with immunoblotting and quantified as described above. The experiment was replicated three times. The data are expressed as the mean ± SEM of three independent experiments. * denotes statistically significant difference from control group (<i>p</i><0.05). <b>B</b>. SH-SY5Y cells were exposed to ethanol (0 or 0.4%) for specified times. After the treatment, cells were lysed and the lysates were immunoprecipitated (IP) with an anti-p47<sup>phox</sup> or anti-p67<sup>phox</sup> antibody. The immunocomplexes were resolved by electrophoresis on a 10% SDS-polyacrylamide gel followed by immunoblotting analysis using either an anti-phosphoserine, anti-p47<sup>phox</sup>, or anti-p67<sup>phox</sup> antibody. <b>C</b>. Cytoplasm and membrane proteins were extracted as described under the <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0038075#s2\" target=\"_blank\">Materials and Methods</a>. The expression of p47<sup>phox</sup> and p67<sup>phox</sup> in the cytoplasm/membrane fractions was determined with immunoblotting. The expression of GAPDH and pan-cadherin served as internal loading controls for cytoplasmic and membrane fractions, respectively. Relative amounts of p47<sup>phox</sup> and p67<sup>phox</sup> was measured by densitometry and normalized to the expression of GAPDH or pan-cadherin. The experiment was replicated three times. The data are expressed as the mean ± SEM of three independent experiments. * denotes statistically significant difference from control group (<i>p</i><0.05).</p>", "links"=>[], "tags"=>["ethanol"], "article_id"=>303546, "categories"=>["Biochemistry", "Neuroscience", "Cell Biology", "Mental Health"], "users"=>["Xin Wang", "Zunji Ke", "Gang Chen", "Mei Xu", "Kimberly A. Bower", "Jacqueline A. Frank", "Zhuo Zhang", "Xianglin Shi", "Jia Luo"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0038075.g002", "stats"=>{"downloads"=>1, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_ethanol_on_p47_phox_and_p67_phox_/303546", "title"=>"Effect of ethanol on p47<sup>phox</sup> and p67<sup>phox</sup>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 04:42:59"}
  • {"files"=>["https://ndownloader.figshare.com/files/633405"], "description"=>"<p><b>A</b>: SH-SY5Y cells were exposed to ethanol (0 or 0.4%) for indicated times. Active Cdc42 was pulled down by PAK1-PBD color agarose beads and analyzed by immunoblotting using an anti-Cdc42 antibody as described under the <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0038075#s2\" target=\"_blank\">Materials and Methods</a> (top panel). Relative expression of active Cdc42 was quantified (bottom panel). The data are expressed as the mean ± SEM of three independent experiments. * denotes statistically significant difference from control cells (<i>p</i><0.05). <b>B</b>. SH-SY5Y cells were transfected with dominant-negative Cdc42 (DN-Cdc42) for 48 hours to inhibit endogenous Cdc42 activity. Active Cdc42 was determined as described above (top panel). After transfection with DN-Cdc42, cells were exposed to ethanol (0.4%, 24 hours). The effect of ethanol on NOX activity and ROS generation was investigated as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0038075#pone-0038075-g001\" target=\"_blank\">Figure 1</a> (bottom panel). The data are expressed as the mean ± SEM of three independent experiments. * denotes statistically significant difference from control cells (<i>p</i><0.05); # denotes statistically significant difference from ethanol-treated cells (<i>p</i><0.05). <b>C</b>. SH-SY5Y cells were transfected with DN-Cdc42 for 48 hours. After transfection, cells were exposed to ethanol (0.4%, 24 hours). The effect of ethanol on p47<sup>phox</sup> phosphorylation was investigated as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0038075#pone-0038075-g002\" target=\"_blank\">Figure 2B</a>. <b>D</b>. SH-SY5Y cells were transfected with DN-Cdc42 for 48 hours. After transfection with DN-Cdc42, cells were exposed to ethanol (0.4%, 24 hours). Cytoplasm and membrane proteins were extracted as described under the <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0038075#s2\" target=\"_blank\">Materials and Methods</a>. The expression of p47<sup>phox</sup> in the cytoplasm/membrane fractions was determined with immunoblotting. The expression of GAPDH and pan-cadherin served as internal loading controls for cytoplasmic and membrane fractions, respectively. Relative amounts of p47<sup>phox</sup> was measured by densitometry and normalized to the expression of GAPDH or pan-cadherin. The experiment was replicated three times. The data are expressed as the mean ± SEM of three independent experiments. * denotes statistically significant difference from control cells (<i>p</i><0.05). # denotes statistically significant difference from ethanol-treated cells (<i>p</i><0.05).</p>", "links"=>[], "tags"=>["cdc42", "ethanol-induced", "nox", "activation", "ros"], "article_id"=>303881, "categories"=>["Biochemistry", "Neuroscience", "Cell Biology", "Mental Health"], "users"=>["Xin Wang", "Zunji Ke", "Gang Chen", "Mei Xu", "Kimberly A. Bower", "Jacqueline A. Frank", "Zhuo Zhang", "Xianglin Shi", "Jia Luo"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0038075.g005", "stats"=>{"downloads"=>2, "page_views"=>34, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Role_of_Cdc42_in_ethanol_induced_NOX_activation_and_ROS_generation_/303881", "title"=>"Role of Cdc42 in ethanol-induced NOX activation and ROS generation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 04:44:45"}

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Relative Metric

{"start_date"=>"2012-01-01T00:00:00Z", "end_date"=>"2012-12-31T00:00:00Z", "subject_areas"=>[{"subject_area"=>"/Biology and life sciences", "average_usage"=>[322, 550, 671, 773, 864, 955, 1048, 1135, 1223, 1308, 1387, 1465, 1534, 1602, 1673, 1744, 1813, 1885, 1955, 2026, 2093, 2160, 2228, 2290, 2349]}, {"subject_area"=>"/Biology and life sciences/Genetics", "average_usage"=>[333, 576, 707, 814, 908, 1004, 1104, 1197, 1280, 1370, 1449, 1531, 1603, 1673, 1742, 1817, 1886, 1954, 2025, 2098, 2171, 2234, 2304, 2365, 2431]}, {"subject_area"=>"/Physical sciences/Chemistry", "average_usage"=>[302, 508, 622, 720, 804, 888, 973, 1054, 1141, 1219, 1299, 1370, 1442, 1511, 1574, 1644, 1711, 1782, 1846, 1911, 1971, 2030, 2097, 2155, 2217]}]}
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