Evidence for the Mitochondrial Lactate Oxidation Complex in Rat Neurons: Demonstration of an Essential Component of Brain Lactate Shuttles
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{"title"=>"Evidence for the mitochondrial lactate oxidation complex in rat neurons: Demonstration of an essential component of brain lactate shuttles", "type"=>"journal", "authors"=>[{"first_name"=>"Takeshi", "last_name"=>"Hashimoto", "scopus_author_id"=>"7404792968"}, {"first_name"=>"Rajaa", "last_name"=>"Hussien", "scopus_author_id"=>"14008043300"}, {"first_name"=>"Hyung Sook", "last_name"=>"Cho", "scopus_author_id"=>"14008377500"}, {"first_name"=>"Daniela", "last_name"=>"Kaufer", "scopus_author_id"=>"15919205000"}, {"first_name"=>"George A.", "last_name"=>"Brooks", "scopus_author_id"=>"7202058198"}], "year"=>2008, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "scopus"=>"2-s2.0-51449120723", "sgr"=>"51449120723", "pui"=>"352320167", "isbn"=>"1932-6203 (Electronic)\\n1932-6203 (Linking)", "pmid"=>"18698340", "doi"=>"10.1371/journal.pone.0002915"}, "id"=>"0937b566-5157-31f9-88a6-a73bd29f026f", "abstract"=>"To evaluate the presence of components of a putative Intracellular Lactate Shuttle (ILS) in neurons, we attempted to determine if monocarboxylate (e.g. lactate) transporter isoforms (MCT1 and -2) and lactate dehydrogenase (LDH) are coexpressed in neuronal mitochondria of rat brains. Immunohistochemical analyses of rat brain cross-sections showed MCT1, MCT2, and LDH to colocalize with the mitochondrial inner membrane marker cytochrome oxidase (COX) in cortical, hippocampal, and thalamic neurons. Immunoblotting after immunoprecipitation (IP) of mitochondria from brain homogenates supported the histochemical observations by demonstrating that COX coprecipitated MCT1, MCT2, and LDH. Additionally, using primary cultures from rat cortex and hippocampus as well as immunohistochemistry and immunocoprecipitation techniques, we demonstrated that MCT2 and LDH are coexpressed in mitochondria of cultured neurons. These findings can be interpreted to mean that, as in skeletal muscle, neurons contain a mitochondrial lactate oxidation complex (mLOC) that has the potential to facilitate both intracellular and cell-cell lactate shuttles in brain.", "link"=>"http://www.mendeley.com/research/evidence-mitochondrial-lactate-oxidation-complex-rat-neurons-demonstration-essential-component-brain", "reader_count"=>107, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>7, "Researcher"=>24, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>29, "Student > Postgraduate"=>2, "Other"=>7, "Student > Master"=>16, "Student > Bachelor"=>14, "Lecturer"=>2, "Professor"=>4}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>7, "Researcher"=>24, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>29, "Student > Postgraduate"=>2, "Other"=>7, "Student > Master"=>16, "Student > Bachelor"=>14, "Lecturer"=>2, "Professor"=>4}, "reader_count_by_subject_area"=>{"Unspecified"=>3, "Engineering"=>2, "Biochemistry, Genetics and Molecular Biology"=>3, "Agricultural and Biological Sciences"=>56, "Medicine and Dentistry"=>16, "Neuroscience"=>12, "Design"=>1, "Sports and Recreations"=>10, "Pharmacology, Toxicology and Pharmaceutical Science"=>1, "Chemistry"=>3}, "reader_count_by_subdiscipline"=>{"Design"=>{"Design"=>1}, "Engineering"=>{"Engineering"=>2}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>16}, "Neuroscience"=>{"Neuroscience"=>12}, "Chemistry"=>{"Chemistry"=>3}, "Sports and Recreations"=>{"Sports and Recreations"=>10}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>56}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}, "Unspecified"=>{"Unspecified"=>3}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}}, "reader_count_by_country"=>{"Canada"=>2, "United States"=>3, "Brazil"=>1, "Italy"=>1, "United Kingdom"=>1, "Australia"=>1, "Spain"=>1}, "group_count"=>3}

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  • {"files"=>["https://ndownloader.figshare.com/files/924246"], "description"=>"<p>Superposition of signals from probes for VDAC (A, green), LDH-H (B, red) and MAP2 (C, blue) showed extensive colocalization throughout the neuron (D, yellow/white). Scale bar = 20 µm.</p>", "links"=>[], "tags"=>["images", "cross-sections", "demonstrating", "mitochondrial", "ldh-h", "hippocampal"], "article_id"=>594689, "categories"=>["Cell Biology", "Physiology", "Neuroscience"], "users"=>["Takeshi Hashimoto", "Rajaa Hussien", "Hyung-Sook Cho", "Daniela Kaufer", "George A. Brooks"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0002915.g006", "stats"=>{"downloads"=>1, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Immunohistochemical_images_of_rat_brain_cross_sections_demonstrating_mitochondrial_LDH_H_in_hippocampal_neurons_/594689", "title"=>"Immunohistochemical images of rat brain cross-sections demonstrating mitochondrial LDH-H in hippocampal neurons.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 09:46:22"}
  • {"files"=>["https://ndownloader.figshare.com/files/924442"], "description"=>"<p>Superposition of signals from probes for LDH (A, green), COX (B, red) and MAP2 (C, blue) showed clear colocalization (D, yellow/white) in neuron cells. Scale bar = 10 µm.</p>", "links"=>[], "tags"=>["images", "demonstrating", "mitochondrial", "ldh", "neurons", "cultured", "hippocampus"], "article_id"=>594886, "categories"=>["Cell Biology", "Physiology", "Neuroscience"], "users"=>["Takeshi Hashimoto", "Rajaa Hussien", "Hyung-Sook Cho", "Daniela Kaufer", "George A. Brooks"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0002915.g009", "stats"=>{"downloads"=>1, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Immunohistochemical_images_demonstrating_mitochondrial_LDH_in_neurons_cultured_from_rat_hippocampus_and_cortex_/594886", "title"=>"Immunohistochemical images demonstrating mitochondrial LDH in neurons cultured from rat hippocampus and cortex.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 09:47:32"}
  • {"files"=>["https://ndownloader.figshare.com/files/924026"], "description"=>"<p>Superposition of signals from probes for MCT2 (A, green), COX (B, red) and MAP2 (C, blue) showed clear colocalization (D, yellow/white) in neurons. Scale bar = 10 µm.</p>", "links"=>[], "tags"=>["images", "cross-sections", "demonstrating", "mitochondrial", "mct2", "hippocampal"], "article_id"=>594473, "categories"=>["Cell Biology", "Physiology", "Neuroscience"], "users"=>["Takeshi Hashimoto", "Rajaa Hussien", "Hyung-Sook Cho", "Daniela Kaufer", "George A. Brooks"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0002915.g003", "stats"=>{"downloads"=>3, "page_views"=>21, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Immunohistochemical_images_of_rat_brain_cross_sections_demonstrating_mitochondrial_MCT2_in_hippocampal_neurons_/594473", "title"=>"Immunohistochemical images of rat brain cross-sections demonstrating mitochondrial MCT2 in hippocampal neurons.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 09:45:00"}
  • {"files"=>["https://ndownloader.figshare.com/files/924585"], "description"=>"<p>Results were compared to those obtained on the supernatant remaining after immunoprecipitation (Sup). COX and nIgG immunoprecipitates were subsequently probed with an antibody that recognized all LDH isoforms. Mitochondrial fractions (MI) were also probed for LDH and results are presented as a positive control. LDH was coprecipitated with COX as compared to nIgG (no protein was coprecipitated with nIgG) in mitochondrial fractions derived from cortical as well as hippocampal areas of rat primary cultures. When the inhibitor to astrocyte development Ara-C was not used, COX coprecipitated LDH in mitochondrial fractions from cortical and hippocampal areas of rat primary cultures. Similar results obtained with and without Ara-C (the lack of Ara-C facilitates development of astrocytes among as neurons in cultures) indicate that astrocytes also contain mLDH.</p>", "links"=>[], "tags"=>["probe", "mitochondrial", "cytochrome", "oxidase", "lactate", "dehydrogenase", "antibodies", "cox", "igg", "immunoprecipitates", "tween", "20", "fractions", "cortex", "hippocampus"], "article_id"=>595018, "categories"=>["Cell Biology", "Physiology", "Neuroscience"], "users"=>["Takeshi Hashimoto", "Rajaa Hussien", "Hyung-Sook Cho", "Daniela Kaufer", "George A. Brooks"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0002915.g011", "stats"=>{"downloads"=>3, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_To_probe_for_an_association_between_mitochondrial_cytochrome_oxidase_COX_and_lactate_dehydrogenase_LDH_antibodies_to_COX_and_normal_IgG_nIgG_were_used_to_develop_immunoprecipitates_IP_from_Tween_20_0_3_solubilized_mitochondrial_fractions_of_cortex_and_hi/595018", "title"=>"To probe for an association between mitochondrial cytochrome oxidase (COX) and lactate dehydrogenase (LDH), antibodies to COX and normal IgG (nIgG) were used to develop immunoprecipitates (IP) from Tween 20 (0.3%)-solubilized mitochondrial fractions of cortex and hippocampus primary cultures.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 09:48:22"}
  • {"files"=>["https://ndownloader.figshare.com/files/923939"], "description"=>"<p>When signals from probes for MCT2 (A, green), and mitochondrial COX (B, red) were merged with those of the neuronal marker MAP2 (C, blue), superposition of the signals (D, yellow/white) showed colocalization of MCT2 and components of the mitochondrial reticulum in neurons (white arrows). Scale bar = 20 µm.</p>", "links"=>[], "tags"=>["images", "cross-sections", "demonstrating", "mitochondrial", "mct2", "thalamic"], "article_id"=>594382, "categories"=>["Cell Biology", "Physiology", "Neuroscience"], "users"=>["Takeshi Hashimoto", "Rajaa Hussien", "Hyung-Sook Cho", "Daniela Kaufer", "George A. Brooks"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0002915.g002", "stats"=>{"downloads"=>2, "page_views"=>21, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Immunohistochemical_images_of_rat_brain_cross_sections_demonstrating_mitochondrial_MCT2_in_thalamic_neurons_/594382", "title"=>"Immunohistochemical images of rat brain cross-sections demonstrating mitochondrial MCT2 in thalamic neurons.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 09:44:31"}
  • {"files"=>["https://ndownloader.figshare.com/files/924097"], "description"=>"<p>In hippocampus (A and D), signals for LDH-H predominate (D, arrows) whereas a signal for LDH-M is not evident (A). On the other hand, in cortex (B and E), signals for LDH-M predominate (B, arrows), but a signal for LDH-H is not evident (E). In thalamus (C and F), signals for LDH-H predominate (F, arrows), but a signal for LDH-M is also apparent (C, arrow). Scale bar = 200 µm.</p>", "links"=>[], "tags"=>["images", "cross-sections", "area-specific", "ldh-m", "ldh-h"], "article_id"=>594543, "categories"=>["Cell Biology", "Physiology", "Neuroscience"], "users"=>["Takeshi Hashimoto", "Rajaa Hussien", "Hyung-Sook Cho", "Daniela Kaufer", "George A. Brooks"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0002915.g004", "stats"=>{"downloads"=>0, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Immunohistochemical_images_of_rat_brain_cross_sections_showing_area_specific_expression_of_LDH_M_A_8211_C_and_LDH_H_D_8211_F_/594543", "title"=>"Immunohistochemical images of rat brain cross-sections showing area-specific expression of LDH-M (A–C) and LDH-H (D–F).", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 09:45:27"}
  • {"files"=>["https://ndownloader.figshare.com/files/924356"], "description"=>"<p>Superposition of signals from probes for MCT2 (A, green), COX (B, red), and MAP2 (C, blue) showed extensive colocalization (D, white) in a cultured neuron. Scale bar = 10 µm.</p>", "links"=>[], "tags"=>["images", "demonstrating", "mitochondrial", "mct2", "cultured", "neuron"], "article_id"=>594804, "categories"=>["Cell Biology", "Physiology", "Neuroscience"], "users"=>["Takeshi Hashimoto", "Rajaa Hussien", "Hyung-Sook Cho", "Daniela Kaufer", "George A. Brooks"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0002915.g008", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Immunohistochemical_images_demonstrating_mitochondrial_MCT2_in_a_cultured_neuron_from_rat_hippocampus_/594804", "title"=>"Immunohistochemical images demonstrating mitochondrial MCT2 in a cultured neuron from rat hippocampus.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 09:47:02"}
  • {"files"=>["https://ndownloader.figshare.com/files/924164"], "description"=>"<p>When signals from probes for LDH-M (B, red) and the mitochondrial protein VDAC (A, green) were merged with those for the neuronal marker MAP2 (C, blue), superposition of the signals (D, yellow/white) showed colocalization of LDH-M and components of the mitochondrial reticulum. Scale bar = 20 µm.</p>", "links"=>[], "tags"=>["images", "demonstrating", "mitochondrial", "ldh-m", "cortical"], "article_id"=>594615, "categories"=>["Cell Biology", "Physiology", "Neuroscience"], "users"=>["Takeshi Hashimoto", "Rajaa Hussien", "Hyung-Sook Cho", "Daniela Kaufer", "George A. Brooks"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0002915.g005", "stats"=>{"downloads"=>1, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Immunohistochemical_images_of_rat_brain_demonstrating_mitochondrial_LDH_M_in_cortical_neurons_/594615", "title"=>"Immunohistochemical images of rat brain demonstrating mitochondrial LDH-M in cortical neurons.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 09:45:53"}
  • {"files"=>["https://ndownloader.figshare.com/files/924303"], "description"=>"<p>IP proteins were subsequently probed with antibodies to MCT1, MCT2, LDH, and β<sub>1</sub>-Na<sup>+</sup>-K<sup>+</sup>-ATPase. MCT1 and MCT2 were coprecipitated with COX, and LDH was slightly, but significantly coprecipitated with COX as compared to nIgG (no protein was coprecipitated with nIgG). The plasma membrane marker β<sub>1</sub>-Na<sup>+</sup>-K<sup>+</sup>-ATPase was not coprecipitated with either COX or nIgG. Sup, supernatant after immunoprecipitation in lysates of mitochondrial fractions.</p>", "links"=>[], "tags"=>["cox", "igg", "immunoprecipitates", "tween", "20", "mitochondrial", "fractions"], "article_id"=>594745, "categories"=>["Cell Biology", "Physiology", "Neuroscience"], "users"=>["Takeshi Hashimoto", "Rajaa Hussien", "Hyung-Sook Cho", "Daniela Kaufer", "George A. Brooks"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0002915.g007", "stats"=>{"downloads"=>1, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Antibodies_to_COX_and_normal_IgG_nIgG_were_used_to_develop_immunoprecipitates_IP_from_Tween_20_1_solubilized_mitochondrial_fractions_of_brain_tissue_/594745", "title"=>"Antibodies to COX and normal IgG (nIgG) were used to develop immunoprecipitates (IP) from Tween 20 (1%)-solubilized mitochondrial fractions of brain tissue.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 09:46:43"}
  • {"files"=>["https://ndownloader.figshare.com/files/923856"], "description"=>"<p>When signals from probes for the lactate/pyruvate transporter MCT1 (A, green) and mitochondrial cytochrome oxidase, COX (B, red) were merged with those of the neuronal marker MAP2 (C, blue), superposition of the signals (D, yellow/white) showed colocalization of MCT1 and components of the mitochondrial reticulum in neurons (white arrows). Scale bar = 20 µm.</p>", "links"=>[], "tags"=>["images", "cross-sections", "demonstrating", "mitochondrial", "mct1", "thalamic"], "article_id"=>594298, "categories"=>["Cell Biology", "Physiology", "Neuroscience"], "users"=>["Takeshi Hashimoto", "Rajaa Hussien", "Hyung-Sook Cho", "Daniela Kaufer", "George A. Brooks"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0002915.g001", "stats"=>{"downloads"=>1, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Immunohistochemical_images_of_rat_brain_cross_sections_demonstrating_mitochondrial_MCT1_in_thalamic_neurons_/594298", "title"=>"Immunohistochemical images of rat brain cross-sections demonstrating mitochondrial MCT1 in thalamic neurons.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 09:43:56"}
  • {"files"=>["https://ndownloader.figshare.com/files/924522"], "description"=>"<p>Whole homogenates (WH) of the same cells were loaded as positive controls. Mitochondria were immunoblotted using plasma membrane markers for β<sub>1</sub>-Na<sup>+</sup>-K<sup>+</sup>-ATPase and GLUT1 as well as the cytosol marker F-actin. Mitochondrial fractions (MI) showed the presence of β<sub>1</sub>-Na<sup>+</sup>-K<sup>+</sup>-ATPase. However, while prominent in whole cell homogenates (WH), the other plasma membrane marker Glut1 and the cytosol marker F-actin were not apparent.</p>", "links"=>[], "tags"=>["mitochondrial", "fractions", "neuronal", "cultures", "developed", "cortex"], "article_id"=>594964, "categories"=>["Cell Biology", "Physiology", "Neuroscience"], "users"=>["Takeshi Hashimoto", "Rajaa Hussien", "Hyung-Sook Cho", "Daniela Kaufer", "George A. Brooks"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0002915.g010", "stats"=>{"downloads"=>1, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Immunoblots_of_mitochondrial_fractions_MI_from_primary_neuronal_cultures_developed_from_rat_cortex_and_hippocampus_/594964", "title"=>"Immunoblots of mitochondrial fractions (MI) from primary neuronal cultures developed from rat cortex and hippocampus.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 09:48:04"}

PMC Usage Stats | Further Information

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Relative Metric

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