Mitochondrial Fragmentation Is Involved in Methamphetamine-Induced Cell Death in Rat Hippocampal Neural Progenitor Cells
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{"title"=>"Mitochondrial fragmentation is involved in methamphetamine-induced cell death in rat hippocampal neural progenitor cells", "type"=>"journal", "authors"=>[{"first_name"=>"Changhai", "last_name"=>"Tian", "scopus_author_id"=>"7102029622"}, {"first_name"=>"L. Charles", "last_name"=>"Murrin", "scopus_author_id"=>"7006267855"}, {"first_name"=>"Jialin C.", "last_name"=>"Zheng", "scopus_author_id"=>"7403974809"}], "year"=>2009, "source"=>"PLoS ONE", "identifiers"=>{"pui"=>"354613568", "issn"=>"19326203", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "doi"=>"10.1371/journal.pone.0005546", "scopus"=>"2-s2.0-65849443539", "pmid"=>"19436752", "sgr"=>"65849443539"}, "id"=>"195026dd-434d-32e0-828e-ec8e776ff2a8", "abstract"=>"Methamphetamine (METH) induces neurodegeneration through damage and apoptosis of dopaminergic nerve terminals and striatal cells, presumably via cross-talk between the endoplasmic reticulum and mitochondria-dependent death cascades. However, the effects of METH on neural progenitor cells (NPC), an important reservoir for replacing neurons and glia during development and injury, remain elusive. Using a rat hippocampal NPC (rhNPC) culture, we characterized the METH-induced mitochondrial fragmentation, apoptosis, and its related signaling mechanism through immunocytochemistry, flow cytometry, and Western blotting. We observed that METH induced rhNPC mitochondrial fragmentation, apoptosis, and inhibited cell proliferation. The mitochondrial fission protein dynamin-related protein 1 (Drp1) and reactive oxygen species (ROS), but not calcium (Ca2+) influx, were involved in the regulation of METH-induced mitochondrial fragmentation. Furthermore, our results indicated that dysregulation of ROS contributed to the oligomerization and translocation of Drp1, resulting in mitochondrial fragmentation in rhNPC. Taken together, our data demonstrate that METH-mediated ROS generation results in the dysregulation of Drp1, which leads to mitochondrial fragmentation and subsequent apoptosis in rhNPC. This provides a potential mechanism for METH-related neurodegenerative disorders, and also provides insight into therapeutic strategies for the neurodegenerative effects of METH.", "link"=>"http://www.mendeley.com/research/mitochondrial-fragmentation-involved-methamphetamineinduced-cell-death-rat-hippocampal-neural-progen-2", "reader_count"=>20, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>1, "Researcher"=>4, "Student > Ph. D. Student"=>10, "Student > Master"=>2, "Student > Bachelor"=>1, "Lecturer"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>1, "Researcher"=>4, "Student > Ph. D. Student"=>10, "Student > Master"=>2, "Student > Bachelor"=>1, "Lecturer"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>1, "Biochemistry, Genetics and Molecular Biology"=>2, "Medicine and Dentistry"=>4, "Agricultural and Biological Sciences"=>9, "Neuroscience"=>2, "Pharmacology, Toxicology and Pharmaceutical Science"=>1, "Immunology and Microbiology"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>4}, "Neuroscience"=>{"Neuroscience"=>2}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>9}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>2}, "Unspecified"=>{"Unspecified"=>1}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}}, "reader_count_by_country"=>{"United States"=>2, "Japan"=>1}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/898116"], "description"=>"<p>Neurospheres were digested and plated on poly-D-lysine-coated cover slips. After 24 hours NPC were treated with 300 µM METH for the indicated times and then subjected to mitochondrial staining with MitoTracker®Red (upper panel) and immunostaining with nestin monoclonal antibody (green) and nuclear staining with DAPI (blue) and visualized by Zeiss Axiovert microscope.</p>", "links"=>[], "tags"=>["induces", "mitochondrial", "fragmentation", "rhnpc", "time-dependent"], "article_id"=>568574, "categories"=>["Neuroscience"], "users"=>["Changhai Tian", "L. Charles Murrin", "Jialin C. Zheng"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0005546.g004", "stats"=>{"downloads"=>1, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_METH_induces_mitochondrial_fragmentation_in_rhNPC_in_a_time_dependent_manner_/568574", "title"=>"METH induces mitochondrial fragmentation in rhNPC in a time-dependent manner.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 05:57:21"}
  • {"files"=>["https://ndownloader.figshare.com/files/897951"], "description"=>"<p>Neurospheres were digested and plated on poly-D-lysine-coated cover slips. After 24 hours rhNPC were treated with METH at indicated concentrations for another 24 hours. (A) NPC were stained with MitoTracker®Red CMXRos dye (red) and fixed with 3.7% formaldehyde. After permeabilization with 0.2% Triton X-100 in PBS at 4°C for 10 min, cells were subjected to immunostaining with monoclonal nestin antibody and nuclei were stained with DAPI (blue), then visualized by Zeiss Axiovert microscope. (B) Apoptotic nuclei, showing highly condensed and fragmented chromatin, as indicated with arrows in A-b2 and A-c2. The results are expressed as average±SD of triplicate samples. # denotes <i>p</i><0.05 in comparison to control; ## denotes <i>p</i><0.001 compared with control. (C) Cells were analyzed for mitochondrial morphology by fluorescence microscope using MitoTracker®Red. Typical mitochondrial phenotypes (A-a1) and fragmented mitochondria (A-b1 and A-c1) were quantified by counting cell number with or without fragmented mitochondria. # denotes <i>p</i><0.01 in comparison to control; ## denotes <i>p</i><0.001 compared with control.</p>", "links"=>[], "tags"=>["apoptosis", "rhnpc", "accompanied", "mitochondrial"], "article_id"=>568411, "categories"=>["Neuroscience"], "users"=>["Changhai Tian", "L. Charles Murrin", "Jialin C. Zheng"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0005546.g003", "stats"=>{"downloads"=>4, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_METH_induced_apoptosis_of_rhNPC_is_accompanied_by_mitochondrial_fragmentation_/568411", "title"=>"METH-induced apoptosis of rhNPC is accompanied by mitochondrial fragmentation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 05:56:28"}
  • {"files"=>["https://ndownloader.figshare.com/files/897740"], "description"=>"<p>rhNPC were dissociated from rat hippocampi and cultured with NPBM neural progenitor basal medium containing EGF, bFGF and NSF-1. The neurospheres were formed after culture for 7 days (Fig. 1A). Neurospheres were digested and plated on poly-D-lysine-coated cover slips and cultured with astrocyte differentiation medium or neuron differentiation medium for 7 days and then subjected to immunostaining with nestin (Fig. 1B), GFAP (Fig. 1C) and β-tubulin III (Fig. 1D) antibodies. rhNPC were cultured in six-well plates with differentiation medium for 3, 5 and 7 days, respectively, then subjected to western blotting with GFAP and β-tubulin III antibodies (Fig. 1E, 1F).</p>", "links"=>[], "tags"=>["differentiation"], "article_id"=>568194, "categories"=>["Neuroscience"], "users"=>["Changhai Tian", "L. Charles Murrin", "Jialin C. Zheng"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0005546.g001", "stats"=>{"downloads"=>1, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Isolation_and_differentiation_of_rhNPC_/568194", "title"=>"Isolation and differentiation of rhNPC.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 05:55:16"}
  • {"files"=>["https://ndownloader.figshare.com/files/898459"], "description"=>"<p>Neurospheres derived from rhNPC were digested and cells were plated on poly-D-lysine-coated cover slips. Cells were treated with 300 µM METH for 24 hours or pre-treated with 10 mM NAC for 30 min, then treated with 300 µM METH for 24 hours. Cells were then subjected to mitochondrial ROS staining with MitoTracker® CM-H<sub>2</sub>ROS (a reduced, nonfluorescent dye that fluoresces upon oxidation) (a2-d2, red), immunostaining with mouse nestin monoclonal antibody (a1-d1, green) and nucleus staining with DAPI (a3-d3, blue) and visualized by Zeiss Axiovert microscope. Cells were treated with 200 µM H<sub>2</sub>O<sub>2</sub> for 30 min as a positive control (e1-f4).</p>", "links"=>[], "tags"=>["increases", "ros"], "article_id"=>568921, "categories"=>["Neuroscience"], "users"=>["Changhai Tian", "L. Charles Murrin", "Jialin C. Zheng"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0005546.g007", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_METH_increases_the_generation_of_ROS_in_rhNPC_/568921", "title"=>"METH increases the generation of ROS in rhNPC.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 05:59:15"}
  • {"files"=>["https://ndownloader.figshare.com/files/898235"], "description"=>"<p>(A) Neurospheres were digested and plated on poly-D-lysine-coated cover slips, and treated with 300 µM METH for 24 hours then subjected to mitochondrial staining with MitoTracker®Red and immunostained with Drp1 monoclonal antibody (green). (B) rhNPC were transfected with synthesized control-siRNA and Drp1-siRNA on poly-D-lysine-coated cover slips, and 48 hours post-transfection, cells were treated with 300 µM METH for another 24 hours and then subjected to mitochondrial staining and immunostaining with Drp1 monoclonal antibody. (C) rhNPC were transfected with synthesized control-siRNA and Drp1-siRNA on poly-D-lysine-coated cover slips, and 48 hours post-transfection cells were subjected to western blot analysis with Drp1 monoclonal antibody, and β-actin was used as a loading control.</p>", "links"=>[], "tags"=>["meth-induced", "mitochondrial", "fragmentation"], "article_id"=>568696, "categories"=>["Neuroscience"], "users"=>["Changhai Tian", "L. Charles Murrin", "Jialin C. Zheng"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0005546.g005", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Drp1_is_involved_in_the_regulation_of_METH_induced_mitochondrial_fragmentation_in_rhNPC_/568696", "title"=>"Drp1 is involved in the regulation of METH-induced mitochondrial fragmentation in rhNPC.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 05:58:02"}
  • {"files"=>["https://ndownloader.figshare.com/files/898368"], "description"=>"<p>rhNPC cultured on poly-D-lysine-coated cover slips (25 mm), were loaded with Fura-2 and monitored for calcium influx by micro-fluorescent imaging. (A) Different concentrations of METH were tested and calcium influx was measured. METH concentrations were 1: 50 µM; 2: 100 µM; 3: 500 µM; 4: 1 mM; 5: 5 mM; 6:100 µM glutamate as a positive control. (B) METH has no effect on glutamate-mediated AMPA receptor responses; a: glutamate (100 µM); b: glutamate (100 µM)+cyclothiazide (10 µM); c: glutamate (100 µM)+cyclothiazide (10 µM)+CNQX (10 µM); d: glutamate (100 µM)+500 µM METH+cyclothiazide (10 µM).</p>", "links"=>[], "tags"=>["influx", "glutamate-mediated"], "article_id"=>568822, "categories"=>["Neuroscience"], "users"=>["Changhai Tian", "L. Charles Murrin", "Jialin C. Zheng"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0005546.g006", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_METH_has_no_effect_on_Ca_2_influx_or_glutamate_mediated_Ca_2_influx_in_rhNPC_/568822", "title"=>"METH has no effect on Ca<sup>2+</sup> influx or glutamate-mediated Ca<sup>2+</sup> influx in rhNPC.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 05:58:42"}
  • {"files"=>["https://ndownloader.figshare.com/files/897857"], "description"=>"<p>Neurospheres were digested and plated on poly-D-lysine-coated six-well plates, and treated with METH at indicated concentrations for 24 hours followed by cell cycle assays. (A) The population of S-phase cells was analyzed by flow cytometry. The results are expressed as average±SD of triplicate samples from three independent experiments. ## denotes <i>p</i><0.05 in comparison to control. (B) rhNPC were pre-treated with 10 mM NAC for 30 min then treated with 300 µM METH for another 24 hours, and the cell cycle was analyzed by flow cytometry. # denotes <i>p</i><0.05 in comparison to control; ## denotes <i>p</i><0.05 compared with METH alone.</p>", "links"=>[], "tags"=>["inhibits", "proliferation"], "article_id"=>568321, "categories"=>["Neuroscience"], "users"=>["Changhai Tian", "L. Charles Murrin", "Jialin C. Zheng"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0005546.g002", "stats"=>{"downloads"=>2, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_METH_inhibits_the_proliferation_of_rhNPC_/568321", "title"=>"METH inhibits the proliferation of rhNPC.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 05:55:57"}
  • {"files"=>["https://ndownloader.figshare.com/files/898657"], "description"=>"<p>(A) rhNPC were treated with different concentrations of METH as indicated, then subjected to non-reducing SDS-PAGE and western blot assay with Drp1 monoclonal antibody and β-actin as a loading control. The data were quantified by ImageJ software (B) ## denotes <i>p</i><0.05 as compared to control. (C) Cells were treated with 300 µM METH or 200 µM H<sub>2</sub>O<sub>2</sub> for 24 hours or were pre-incubated with 10 mM NAC for 30 min then with 300 µM METH or 200 µM H<sub>2</sub>O<sub>2</sub> treatment for 24 hours. They were then subjected to non-reducing SDS-PAGE and western blot assay with Drp1 monoclonal antibody and β-actin as a loading control. (D) Cells were treated with 300 µM METH alone or pre-incubated with 10 mM NAC for 30 min, then with 300 µM METH treatment for 24 hours, and subsequently subjected to cell fractionation and western blotting with Drp1 monoclonal antibody as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0005546#s4\" target=\"_blank\">materials and methods</a>. VDAC1 was used as a mitochondrial marker and loading control.</p>", "links"=>[], "tags"=>["stress-mediated", "oligomerization", "translocation", "drp1", "mitochondrial"], "article_id"=>569113, "categories"=>["Neuroscience"], "users"=>["Changhai Tian", "L. Charles Murrin", "Jialin C. Zheng"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0005546.g008", "stats"=>{"downloads"=>3, "page_views"=>92, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Oxidative_stress_mediated_oligomerization_and_translocation_of_Drp1_contribute_to_mitochondrial_fragmentation_/569113", "title"=>"Oxidative stress-mediated oligomerization and translocation of Drp1 contribute to mitochondrial fragmentation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 06:00:20"}

PMC Usage Stats | Further Information

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  • {"unique-ip"=>"4", "full-text"=>"2", "pdf"=>"2", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"4", "supp-data"=>"0", "cited-by"=>"1", "year"=>"2017", "month"=>"1"}
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  • {"unique-ip"=>"2", "full-text"=>"2", "pdf"=>"5", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"1"}
  • {"unique-ip"=>"10", "full-text"=>"5", "pdf"=>"7", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"3"}
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  • {"unique-ip"=>"5", "full-text"=>"5", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"9"}
  • {"unique-ip"=>"4", "full-text"=>"4", "pdf"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"12"}
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  • {"unique-ip"=>"7", "full-text"=>"7", "pdf"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"2"}
  • {"unique-ip"=>"5", "full-text"=>"6", "pdf"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"3"}
  • {"unique-ip"=>"3", "full-text"=>"4", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"4"}
  • {"unique-ip"=>"5", "full-text"=>"9", "pdf"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"5"}
  • {"unique-ip"=>"7", "full-text"=>"8", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"8"}
  • {"unique-ip"=>"8", "full-text"=>"6", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"9"}
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Relative Metric

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