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Stoichiometry of δ subunit containing GABAA receptors
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| Zeitschriftentitel: | British Journal of Pharmacology |
|---|---|
| Personen und Körperschaften: | , , |
| In: | British Journal of Pharmacology, 171, 2014, 4, S. 985-994 |
| Format: | E-Article |
| Sprache: | Englisch |
| veröffentlicht: |
Wiley
|
| Schlagwörter: |
| author_facet |
Patel, B Mortensen, M Smart, T G Patel, B Mortensen, M Smart, T G |
|---|---|
| author |
Patel, B Mortensen, M Smart, T G |
| spellingShingle |
Patel, B Mortensen, M Smart, T G British Journal of Pharmacology Stoichiometry of δ subunit containing GABAA receptors Pharmacology |
| author_sort |
patel, b |
| spelling |
Patel, B Mortensen, M Smart, T G 0007-1188 1476-5381 Wiley Pharmacology http://dx.doi.org/10.1111/bph.12514 <jats:sec><jats:title>Background and Purpose</jats:title><jats:p>Although the stoichiometry of the major synaptic αβγ subunit‐containing <jats:styled-content style="fixed-case">GABA<jats:sub>A</jats:sub></jats:styled-content> receptors has consensus support for 2α:2β:1γ, a clear view of the stoichiometry of extrasynaptic receptors containing δ subunits has remained elusive. Here we examine the subunit stoichiometry of recombinant α4β3δ receptors using a reporter mutation and a functional electrophysiological approach.</jats:p></jats:sec><jats:sec><jats:title>Experimental Approach</jats:title><jats:p>Using site‐directed mutagenesis, we inserted a highly characterized 9′ serine to leucine mutation into the second transmembrane (<jats:styled-content style="fixed-case">M</jats:styled-content>2) region of α4, β3 and δ subunits that increases receptor sensitivity to <jats:styled-content style="fixed-case">GABA</jats:styled-content>. Whole‐cell, <jats:styled-content style="fixed-case">GABA</jats:styled-content>‐activated currents were recorded from <jats:styled-content style="fixed-case">HEK</jats:styled-content>‐293 cells co‐expressing different combinations of wild‐type (<jats:styled-content style="fixed-case">WT</jats:styled-content>) and/or mutant α4(<jats:styled-content style="fixed-case">L</jats:styled-content>297<jats:styled-content style="fixed-case">S</jats:styled-content>), β3(<jats:styled-content style="fixed-case">L</jats:styled-content>284<jats:styled-content style="fixed-case">S</jats:styled-content>) and δ(<jats:styled-content style="fixed-case">L</jats:styled-content>288<jats:styled-content style="fixed-case">S</jats:styled-content>) subunits.</jats:p></jats:sec><jats:sec><jats:title>Key Results</jats:title><jats:p>Recombinant receptors containing one or more mutant subunits showed increased <jats:styled-content style="fixed-case">GABA</jats:styled-content> sensitivity relative to <jats:styled-content style="fixed-case">WT</jats:styled-content> receptors by approximately fourfold, independent of the subunit class (α, β or δ) carrying the mutation. <jats:styled-content style="fixed-case">GABA</jats:styled-content> dose–response curves of cells co‐expressing <jats:styled-content style="fixed-case">WT</jats:styled-content> subunits with their respective <jats:styled-content style="fixed-case">L</jats:styled-content>9′<jats:styled-content style="fixed-case">S</jats:styled-content> mutants exhibited multiple components, with the number of discernible components enabling a subunit stoichiometry of 2α, 2β and 1δ to be deduced for α4β3δ receptors. Varying the <jats:styled-content style="fixed-case">cDNA</jats:styled-content> transfection ratio by 10‐fold had no significant effect on the number of incorporated δ subunits.</jats:p></jats:sec><jats:sec><jats:title>Conclusions and Implications</jats:title><jats:p>Subunit stoichiometry is an important determinant of <jats:styled-content style="fixed-case">GABA<jats:sub>A</jats:sub></jats:styled-content> receptor function and pharmacology, and δ subunit‐containing receptors are important mediators of tonic inhibition in several brain regions. Here we demonstrate a preferred subunit stoichiometry for α4β3δ receptors of 2α, 2β and 1δ.</jats:p></jats:sec> Stoichiometry of δ subunit containing <scp>GABA<sub>A</sub></scp> receptors British Journal of Pharmacology |
| doi_str_mv |
10.1111/bph.12514 |
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Online Free |
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Chemie und Pharmazie |
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ElectronicArticle |
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| imprint |
Wiley, 2014 |
| imprint_str_mv |
Wiley, 2014 |
| issn |
0007-1188 1476-5381 |
| issn_str_mv |
0007-1188 1476-5381 |
| language |
English |
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Wiley (CrossRef) |
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patel2014stoichiometryofdsubunitcontaininggabaareceptors |
| publishDateSort |
2014 |
| publisher |
Wiley |
| recordtype |
ai |
| record_format |
ai |
| series |
British Journal of Pharmacology |
| source_id |
49 |
| title |
Stoichiometry of δ subunit containing GABAA receptors |
| title_unstemmed |
Stoichiometry of δ subunit containing GABAA receptors |
| title_full |
Stoichiometry of δ subunit containing GABAA receptors |
| title_fullStr |
Stoichiometry of δ subunit containing GABAA receptors |
| title_full_unstemmed |
Stoichiometry of δ subunit containing GABAA receptors |
| title_short |
Stoichiometry of δ subunit containing GABAA receptors |
| title_sort |
stoichiometry of δ subunit containing <scp>gaba<sub>a</sub></scp> receptors |
| topic |
Pharmacology |
| url |
http://dx.doi.org/10.1111/bph.12514 |
| publishDate |
2014 |
| physical |
985-994 |
| description |
<jats:sec><jats:title>Background and Purpose</jats:title><jats:p>Although the stoichiometry of the major synaptic αβγ subunit‐containing <jats:styled-content style="fixed-case">GABA<jats:sub>A</jats:sub></jats:styled-content> receptors has consensus support for 2α:2β:1γ, a clear view of the stoichiometry of extrasynaptic receptors containing δ subunits has remained elusive. Here we examine the subunit stoichiometry of recombinant α4β3δ receptors using a reporter mutation and a functional electrophysiological approach.</jats:p></jats:sec><jats:sec><jats:title>Experimental Approach</jats:title><jats:p>Using site‐directed mutagenesis, we inserted a highly characterized 9′ serine to leucine mutation into the second transmembrane (<jats:styled-content style="fixed-case">M</jats:styled-content>2) region of α4, β3 and δ subunits that increases receptor sensitivity to <jats:styled-content style="fixed-case">GABA</jats:styled-content>. Whole‐cell, <jats:styled-content style="fixed-case">GABA</jats:styled-content>‐activated currents were recorded from <jats:styled-content style="fixed-case">HEK</jats:styled-content>‐293 cells co‐expressing different combinations of wild‐type (<jats:styled-content style="fixed-case">WT</jats:styled-content>) and/or mutant α4(<jats:styled-content style="fixed-case">L</jats:styled-content>297<jats:styled-content style="fixed-case">S</jats:styled-content>), β3(<jats:styled-content style="fixed-case">L</jats:styled-content>284<jats:styled-content style="fixed-case">S</jats:styled-content>) and δ(<jats:styled-content style="fixed-case">L</jats:styled-content>288<jats:styled-content style="fixed-case">S</jats:styled-content>) subunits.</jats:p></jats:sec><jats:sec><jats:title>Key Results</jats:title><jats:p>Recombinant receptors containing one or more mutant subunits showed increased <jats:styled-content style="fixed-case">GABA</jats:styled-content> sensitivity relative to <jats:styled-content style="fixed-case">WT</jats:styled-content> receptors by approximately fourfold, independent of the subunit class (α, β or δ) carrying the mutation. <jats:styled-content style="fixed-case">GABA</jats:styled-content> dose–response curves of cells co‐expressing <jats:styled-content style="fixed-case">WT</jats:styled-content> subunits with their respective <jats:styled-content style="fixed-case">L</jats:styled-content>9′<jats:styled-content style="fixed-case">S</jats:styled-content> mutants exhibited multiple components, with the number of discernible components enabling a subunit stoichiometry of 2α, 2β and 1δ to be deduced for α4β3δ receptors. Varying the <jats:styled-content style="fixed-case">cDNA</jats:styled-content> transfection ratio by 10‐fold had no significant effect on the number of incorporated δ subunits.</jats:p></jats:sec><jats:sec><jats:title>Conclusions and Implications</jats:title><jats:p>Subunit stoichiometry is an important determinant of <jats:styled-content style="fixed-case">GABA<jats:sub>A</jats:sub></jats:styled-content> receptor function and pharmacology, and δ subunit‐containing receptors are important mediators of tonic inhibition in several brain regions. Here we demonstrate a preferred subunit stoichiometry for α4β3δ receptors of 2α, 2β and 1δ.</jats:p></jats:sec> |
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| author | Patel, B, Mortensen, M, Smart, T G |
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| author_sort | patel, b |
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| description | <jats:sec><jats:title>Background and Purpose</jats:title><jats:p>Although the stoichiometry of the major synaptic αβγ subunit‐containing <jats:styled-content style="fixed-case">GABA<jats:sub>A</jats:sub></jats:styled-content> receptors has consensus support for 2α:2β:1γ, a clear view of the stoichiometry of extrasynaptic receptors containing δ subunits has remained elusive. Here we examine the subunit stoichiometry of recombinant α4β3δ receptors using a reporter mutation and a functional electrophysiological approach.</jats:p></jats:sec><jats:sec><jats:title>Experimental Approach</jats:title><jats:p>Using site‐directed mutagenesis, we inserted a highly characterized 9′ serine to leucine mutation into the second transmembrane (<jats:styled-content style="fixed-case">M</jats:styled-content>2) region of α4, β3 and δ subunits that increases receptor sensitivity to <jats:styled-content style="fixed-case">GABA</jats:styled-content>. Whole‐cell, <jats:styled-content style="fixed-case">GABA</jats:styled-content>‐activated currents were recorded from <jats:styled-content style="fixed-case">HEK</jats:styled-content>‐293 cells co‐expressing different combinations of wild‐type (<jats:styled-content style="fixed-case">WT</jats:styled-content>) and/or mutant α4(<jats:styled-content style="fixed-case">L</jats:styled-content>297<jats:styled-content style="fixed-case">S</jats:styled-content>), β3(<jats:styled-content style="fixed-case">L</jats:styled-content>284<jats:styled-content style="fixed-case">S</jats:styled-content>) and δ(<jats:styled-content style="fixed-case">L</jats:styled-content>288<jats:styled-content style="fixed-case">S</jats:styled-content>) subunits.</jats:p></jats:sec><jats:sec><jats:title>Key Results</jats:title><jats:p>Recombinant receptors containing one or more mutant subunits showed increased <jats:styled-content style="fixed-case">GABA</jats:styled-content> sensitivity relative to <jats:styled-content style="fixed-case">WT</jats:styled-content> receptors by approximately fourfold, independent of the subunit class (α, β or δ) carrying the mutation. <jats:styled-content style="fixed-case">GABA</jats:styled-content> dose–response curves of cells co‐expressing <jats:styled-content style="fixed-case">WT</jats:styled-content> subunits with their respective <jats:styled-content style="fixed-case">L</jats:styled-content>9′<jats:styled-content style="fixed-case">S</jats:styled-content> mutants exhibited multiple components, with the number of discernible components enabling a subunit stoichiometry of 2α, 2β and 1δ to be deduced for α4β3δ receptors. Varying the <jats:styled-content style="fixed-case">cDNA</jats:styled-content> transfection ratio by 10‐fold had no significant effect on the number of incorporated δ subunits.</jats:p></jats:sec><jats:sec><jats:title>Conclusions and Implications</jats:title><jats:p>Subunit stoichiometry is an important determinant of <jats:styled-content style="fixed-case">GABA<jats:sub>A</jats:sub></jats:styled-content> receptor function and pharmacology, and δ subunit‐containing receptors are important mediators of tonic inhibition in several brain regions. Here we demonstrate a preferred subunit stoichiometry for α4β3δ receptors of 2α, 2β and 1δ.</jats:p></jats:sec> |
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| spelling | Patel, B Mortensen, M Smart, T G 0007-1188 1476-5381 Wiley Pharmacology http://dx.doi.org/10.1111/bph.12514 <jats:sec><jats:title>Background and Purpose</jats:title><jats:p>Although the stoichiometry of the major synaptic αβγ subunit‐containing <jats:styled-content style="fixed-case">GABA<jats:sub>A</jats:sub></jats:styled-content> receptors has consensus support for 2α:2β:1γ, a clear view of the stoichiometry of extrasynaptic receptors containing δ subunits has remained elusive. Here we examine the subunit stoichiometry of recombinant α4β3δ receptors using a reporter mutation and a functional electrophysiological approach.</jats:p></jats:sec><jats:sec><jats:title>Experimental Approach</jats:title><jats:p>Using site‐directed mutagenesis, we inserted a highly characterized 9′ serine to leucine mutation into the second transmembrane (<jats:styled-content style="fixed-case">M</jats:styled-content>2) region of α4, β3 and δ subunits that increases receptor sensitivity to <jats:styled-content style="fixed-case">GABA</jats:styled-content>. Whole‐cell, <jats:styled-content style="fixed-case">GABA</jats:styled-content>‐activated currents were recorded from <jats:styled-content style="fixed-case">HEK</jats:styled-content>‐293 cells co‐expressing different combinations of wild‐type (<jats:styled-content style="fixed-case">WT</jats:styled-content>) and/or mutant α4(<jats:styled-content style="fixed-case">L</jats:styled-content>297<jats:styled-content style="fixed-case">S</jats:styled-content>), β3(<jats:styled-content style="fixed-case">L</jats:styled-content>284<jats:styled-content style="fixed-case">S</jats:styled-content>) and δ(<jats:styled-content style="fixed-case">L</jats:styled-content>288<jats:styled-content style="fixed-case">S</jats:styled-content>) subunits.</jats:p></jats:sec><jats:sec><jats:title>Key Results</jats:title><jats:p>Recombinant receptors containing one or more mutant subunits showed increased <jats:styled-content style="fixed-case">GABA</jats:styled-content> sensitivity relative to <jats:styled-content style="fixed-case">WT</jats:styled-content> receptors by approximately fourfold, independent of the subunit class (α, β or δ) carrying the mutation. <jats:styled-content style="fixed-case">GABA</jats:styled-content> dose–response curves of cells co‐expressing <jats:styled-content style="fixed-case">WT</jats:styled-content> subunits with their respective <jats:styled-content style="fixed-case">L</jats:styled-content>9′<jats:styled-content style="fixed-case">S</jats:styled-content> mutants exhibited multiple components, with the number of discernible components enabling a subunit stoichiometry of 2α, 2β and 1δ to be deduced for α4β3δ receptors. Varying the <jats:styled-content style="fixed-case">cDNA</jats:styled-content> transfection ratio by 10‐fold had no significant effect on the number of incorporated δ subunits.</jats:p></jats:sec><jats:sec><jats:title>Conclusions and Implications</jats:title><jats:p>Subunit stoichiometry is an important determinant of <jats:styled-content style="fixed-case">GABA<jats:sub>A</jats:sub></jats:styled-content> receptor function and pharmacology, and δ subunit‐containing receptors are important mediators of tonic inhibition in several brain regions. Here we demonstrate a preferred subunit stoichiometry for α4β3δ receptors of 2α, 2β and 1δ.</jats:p></jats:sec> Stoichiometry of δ subunit containing <scp>GABA<sub>A</sub></scp> receptors British Journal of Pharmacology |
| spellingShingle | Patel, B, Mortensen, M, Smart, T G, British Journal of Pharmacology, Stoichiometry of δ subunit containing GABAA receptors, Pharmacology |
| title | Stoichiometry of δ subunit containing GABAA receptors |
| title_full | Stoichiometry of δ subunit containing GABAA receptors |
| title_fullStr | Stoichiometry of δ subunit containing GABAA receptors |
| title_full_unstemmed | Stoichiometry of δ subunit containing GABAA receptors |
| title_short | Stoichiometry of δ subunit containing GABAA receptors |
| title_sort | stoichiometry of δ subunit containing <scp>gaba<sub>a</sub></scp> receptors |
| title_unstemmed | Stoichiometry of δ subunit containing GABAA receptors |
| topic | Pharmacology |
| url | http://dx.doi.org/10.1111/bph.12514 |