Homer1a-Dependent Crosstalk Between NMDA and Metabotropic Glutamate Receptors in Mouse Neurons
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{"title"=>"Homer1a-Dependent Crosstalk Between NMDA and Metabotropic Glutamate Receptors in Mouse Neurons", "type"=>"journal", "authors"=>[{"first_name"=>"Federica", "last_name"=>"Bertaso", "scopus_author_id"=>"6602577784"}, {"first_name"=>"Gautier", "last_name"=>"Roussignol", "scopus_author_id"=>"8353115600"}, {"first_name"=>"Paul", "last_name"=>"Worley", "scopus_author_id"=>"7004403530"}, {"first_name"=>"Joël", "last_name"=>"Bockaert", "scopus_author_id"=>"36064556300"}, {"first_name"=>"Laurent", "last_name"=>"Fagni", "scopus_author_id"=>"56277672600"}, {"first_name"=>"Fabrice", "last_name"=>"Ango", "scopus_author_id"=>"6603392696"}], "year"=>2010, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "scopus"=>"2-s2.0-77955601200", "sgr"=>"77955601200", "pui"=>"368261742", "isbn"=>"1932-6203 (Electronic)\\n1932-6203 (Linking)", "pmid"=>"20305784", "doi"=>"10.1371/journal.pone.0009755"}, "id"=>"2f505487-89b0-35a2-9b4a-2d1beff0b2d0", "abstract"=>"BACKGROUND: A large number of evidences suggest that group-I metabotropic glutamate receptors (mGluR1a, 1b, 1c, 5a, 5b) can modulate NMDA receptor activity. Interestingly, a physical link exists between these receptors through a Homer-Shank multi-protein scaffold that can be disrupted by the immediate early gene, Homer1a. Whether such a versatile link supports functional crosstalk between the receptors is unknown.\\n\\nMETHODOLOGY/PRINCIPAL FINDINGS: Here we used biochemical, electrophysiological and molecular biological approaches in cultured mouse cerebellar neurons to investigate this issue. We found that Homer1a or dominant negative Shank3 mutants that disrupt the physical link between the receptors allow inhibition of NMDA current by group-I mGluR agonist. This effect is antagonized by pertussis toxin, but not thapsigargin, suggesting the involvement of a G protein, but not intracellular calcium stores. Also, this effect is voltage-sensitive, being present at negative, but not positive membrane potentials. In the presence of DHPG, an apparent NMDA \"tail current\" was evoked by large pulse depolarization, only in neurons transfected with Homer1a. Co-immunoprecipitation experiments showed interaction between G-protein betagamma subunits and NMDA receptor in the presence of Homer1a and group-I mGluR agonist.\\n\\nCONCLUSIONS/SIGNIFICANCE: Altogether these results suggest a direct inhibition of NMDA receptor-channel by Gbetagamma subunits, following disruption of the Homer-Shank3 complex by the immediate early gene Homer1a. This study provides a new molecular mechanism by which group-I mGluRs could dynamically regulate NMDA receptor function.", "link"=>"http://www.mendeley.com/research/homer1adependent-crosstalk-between-nmda-metabotropic-glutamate-receptors-mouse-neurons", "reader_count"=>78, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>2, "Researcher"=>19, "Student > Doctoral Student"=>3, "Student > Ph. D. Student"=>27, "Student > Postgraduate"=>9, "Student > Master"=>8, "Student > Bachelor"=>8, "Professor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>2, "Researcher"=>19, "Student > Doctoral Student"=>3, "Student > Ph. D. Student"=>27, "Student > Postgraduate"=>9, "Student > Master"=>8, "Student > Bachelor"=>8, "Professor"=>1}, "reader_count_by_subject_area"=>{"Engineering"=>1, "Unspecified"=>2, "Biochemistry, Genetics and Molecular Biology"=>4, "Medicine and Dentistry"=>5, "Agricultural and Biological Sciences"=>44, "Neuroscience"=>18, "Pharmacology, Toxicology and Pharmaceutical Science"=>2, "Psychology"=>2}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>5}, "Neuroscience"=>{"Neuroscience"=>18}, "Psychology"=>{"Psychology"=>2}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>44}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>4}, "Unspecified"=>{"Unspecified"=>2}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>2}}, "reader_count_by_country"=>{"United States"=>3, "Chile"=>3, "France"=>1, "Spain"=>1}, "group_count"=>1}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/858137"], "description"=>"<p><b>A</b>: Current-voltage relationships obtained with applications of NMDA alone (filled circles) or co-applications of NMDA and DHPG (open circles), in control neurons (a) and neurons transfected with Homer1a (b). <b>B</b>: NMDA currents recorded in neurons transfected with Homer1a. The upper traces illustrate the voltage protocol applied to the neurons. Each lower trace is the expanded part of the middle trace (current) during the depolarizing pulse. The arrow in b indicates the NMDA “tail current”. Panels a and b were obtained in the absence of PTX treatment, while panel c was obtained in a PTX treated neuron. <b>C</b>: Quantification of the inhibitory effect of DHPG on NMDA current recorded before (filled bars; identical to those illustrated in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0009755#pone-0009755-g001\" target=\"_blank\">Fig. 1E</a>) and immediately after (open bars; NMDA “tail current” measured from baseline ( = 0 pA current)) the depolarization pulse, as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0009755#pone-0009755-g001\" target=\"_blank\">Fig. 1E</a>. Each value of the histogram is the mean (± s.e.m.) of at least 10 experiments.</p>", "links"=>[], "tags"=>["nmda-r", "mglur"], "article_id"=>528593, "categories"=>["Neuroscience"], "users"=>["Federica Bertaso", "Gautier Roussignol", "Paul Worley", "Joël Bockaert", "Laurent Fagni", "Fabrice Ango"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0009755.g003", "stats"=>{"downloads"=>1, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Voltage_dependency_of_NMDA_R_and_mGluR_crosstalk_/528593", "title"=>"Voltage-dependency of NMDA-R and mGluR crosstalk.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-03-18 02:23:13"}
  • {"files"=>["https://ndownloader.figshare.com/files/426580"], "description"=>"<div><h3>Background</h3><p>A large number of evidences suggest that group-I metabotropic glutamate receptors (mGluR1a, 1b, 1c, 5a, 5b) can modulate NMDA receptor activity. Interestingly, a physical link exists between these receptors through a Homer-Shank multi-protein scaffold that can be disrupted by the immediate early gene, Homer1a. Whether such a versatile link supports functional crosstalk between the receptors is unknown.</p><h3>Methodology/Principal Findings</h3><p>Here we used biochemical, electrophysiological and molecular biological approaches in cultured mouse cerebellar neurons to investigate this issue. We found that Homer1a or dominant negative Shank3 mutants that disrupt the physical link between the receptors allow inhibition of NMDA current by group-I mGluR agonist. This effect is antagonized by pertussis toxin, but not thapsigargin, suggesting the involvement of a G protein, but not intracellular calcium stores. Also, this effect is voltage-sensitive, being present at negative, but not positive membrane potentials. In the presence of DHPG, an apparent NMDA “tail current” was evoked by large pulse depolarization, only in neurons transfected with Homer1a. Co-immunoprecipitation experiments showed interaction between G-protein βγ subunits and NMDA receptor in the presence of Homer1a and group-I mGluR agonist.</p><h3>Conclusions/Significance</h3><p>Altogether these results suggest a direct inhibition of NMDA receptor-channel by Gbetagamma subunits, following disruption of the Homer-Shank3 complex by the immediate early gene Homer1a. This study provides a new molecular mechanism by which group-I mGluRs could dynamically regulate NMDA receptor function.</p></div>", "links"=>[], "tags"=>["homer1a-dependent", "crosstalk", "nmda", "metabotropic", "glutamate", "receptors", "neurons"], "article_id"=>144202, "categories"=>["Neuroscience"], "users"=>["Federica Bertaso", "Gautier Roussignol", "Paul Worley", "Joël Bockaert", "Laurent Fagni", "Fabrice Ango"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0009755", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Homer1a_Dependent_Crosstalk_Between_NMDA_and_Metabotropic_Glutamate_Receptors_in_Mouse_Neurons/144202", "title"=>"Homer1a-Dependent Crosstalk Between NMDA and Metabotropic Glutamate Receptors in Mouse Neurons", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-03-18 01:10:02"}
  • {"files"=>["https://ndownloader.figshare.com/files/858212"], "description"=>"<p><b>A</b>: GTPgammaS was dialyzed in cultured cerebellar granule cells through the recording pipette and NMDA currents measured before and after a depolarization pulse, as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0009755#pone-0009755-g003\" target=\"_blank\">Fig. 3Ba</a> and <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0009755#pone-0009755-g003\" target=\"_blank\">Fig. 3C</a>. <b>B</b>: Control neurons (- Homer1a) or neurons expressing induced Homer1a (+ Homer1a) were exposed (+) or not (−) to DHPG, plus NMDA. Lysates were prepared from these cultures and immunoprecipitated with a pan anti-NR1 antibody, and revealed with an anti-Gbeta antibody.</p>", "links"=>[], "tags"=>["inhibit", "nmda"], "article_id"=>528667, "categories"=>["Neuroscience"], "users"=>["Federica Bertaso", "Gautier Roussignol", "Paul Worley", "Joël Bockaert", "Laurent Fagni", "Fabrice Ango"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0009755.g004", "stats"=>{"downloads"=>1, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_G_proteins_interact_with_and_inhibit_NMDA_channels_/528667", "title"=>"G-proteins interact with and inhibit NMDA channels.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-03-18 02:24:27"}
  • {"files"=>["https://ndownloader.figshare.com/files/858068"], "description"=>"<p><b>A</b>: NMDA currents recorded in a neuron transfected with the indicated Shank3 mutant and pre-treated for 30 min with thapsigargin. <b>B</b>: Quantification of the inhibitory effect of DHPG on NMDA currents as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0009755#pone-0009755-g001\" target=\"_blank\">Fig. 1E</a>, in control neurons and neurons transfected with the indicated plasmids, and treated with thapsigargin. <b>C</b>: Same legend as in B, but in neurons pre-treated with PTX.</p>", "links"=>[], "tags"=>["nmda-r", "mglur", "g-protein", "intracellular"], "article_id"=>528519, "categories"=>["Neuroscience"], "users"=>["Federica Bertaso", "Gautier Roussignol", "Paul Worley", "Joël Bockaert", "Laurent Fagni", "Fabrice Ango"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0009755.g002", "stats"=>{"downloads"=>1, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Crosstalk_between_NMDA_R_and_mGluR_is_dependent_on_G_protein_activation_but_independent_of_intracellular_Ca_2_release_/528519", "title"=>"Crosstalk between NMDA-R and mGluR is dependent on G-protein activation, but independent of intracellular Ca<sup>2+</sup> release.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-03-18 02:21:59"}
  • {"files"=>["https://ndownloader.figshare.com/files/857957"], "description"=>"<p><b>A–D</b>: NMDA currents recorded in control neurons (A), in neurons transfected with Homer1a (B), Shank3 wild-type (C), or the indicated Shank3 point mutant (D). <b>E</b>: The inhibitory effect of DHPG was quantified by plotting the amplitude of the current evoked by co-application of NMDA and DHPG over the amplitude of the current evoked by NMDA alone, in the same cell. Each value of the histogram is the mean (± s.e.m.) of 10 experiments. <b>F</b>: A brief application of DHPG was performed during the application of NMDA, in a Homer1a transfected neuron, showing fast onset and reversibility of its inhibitory effect on NMDA current. Similar results were observed in all the recorded neurons (n = 10). In this and the following figures, all the currents were recorded at a holding potential of −80 mV.</p>", "links"=>[], "tags"=>["nmda", "currents", "disruption", "mglur1a-homer-shank3"], "article_id"=>528407, "categories"=>["Neuroscience"], "users"=>["Federica Bertaso", "Gautier Roussignol", "Paul Worley", "Joël Bockaert", "Laurent Fagni", "Fabrice Ango"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0009755.g001", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Inhibition_of_NMDA_currents_after_disruption_of_the_mGluR1a_Homer_Shank3_complex_/528407", "title"=>"Inhibition of NMDA currents after disruption of the mGluR1a-Homer-Shank3 complex.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-03-18 02:20:07"}

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