Brain Region Differences in α1- and α5-Subunit-Containing GABA<sub<A</sub< Receptor Proteomes Revealed with Affinity Purification and Blue Native PAGE Proteomics
GABA<sub<A</sub< receptors are the major inhibitory receptors in the brain. They are hetero-pentamers with a composition of predominantly two α, two β, and one γ or δ subunit. Of the six α subunit genes, the α5 subunit displays a limited spatial expression pattern and is known to mediate...
Ausführliche Beschreibung
Autor*in: |
Miao Chen [verfasserIn] Frank Koopmans [verfasserIn] Miguel A. Gonzalez-Lozano [verfasserIn] August B. Smit [verfasserIn] Ka Wan Li [verfasserIn] |
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Format: |
E-Artikel |
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Sprache: |
Englisch |
Erschienen: |
2023 |
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Übergeordnetes Werk: |
In: Cells - MDPI AG, 2012, 13(2023), 1, p 14 |
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Übergeordnetes Werk: |
volume:13 ; year:2023 ; number:1, p 14 |
Links: |
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DOI / URN: |
10.3390/cells13010014 |
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Katalog-ID: |
DOAJ097807907 |
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10.3390/cells13010014 doi (DE-627)DOAJ097807907 (DE-599)DOAJd7f9b4d52a7645c69f1d564e94edeec5 DE-627 ger DE-627 rakwb eng QH573-671 Miao Chen verfasserin aut Brain Region Differences in α1- and α5-Subunit-Containing GABA<sub<A</sub< Receptor Proteomes Revealed with Affinity Purification and Blue Native PAGE Proteomics 2023 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier GABA<sub<A</sub< receptors are the major inhibitory receptors in the brain. They are hetero-pentamers with a composition of predominantly two α, two β, and one γ or δ subunit. Of the six α subunit genes, the α5 subunit displays a limited spatial expression pattern and is known to mediate both phasic and tonic inhibition. In this study, using immunoaffinity-based proteomics, we identified the α5 subunit containing receptor complexes in the hippocampus and olfactory bulb. The α1–α5 interaction was identified in both brain regions, albeit with significantly different stoichiometries. In line with this, reverse IPs using anti-α1 antibodies showed the α5–α1 co-occurrence and validated the quantitative difference. In addition, we showed that the association of Neuroligin 2 with α1-containing receptors was much higher in the olfactory bulb than in the hippocampus, which was confirmed using blue native gel electrophoresis and quantitative mass spectrometry. Finally, immunocytochemical staining revealed a co-localization of α1 and α5 subunits in the post-synaptic puncta in the hippocampus. GABA<sub<A</sub< receptor immunoprecipitation proteomics brain synapse immunocytochemistry Cytology Frank Koopmans verfasserin aut Miguel A. Gonzalez-Lozano verfasserin aut August B. Smit verfasserin aut Ka Wan Li verfasserin aut In Cells MDPI AG, 2012 13(2023), 1, p 14 (DE-627)718622081 (DE-600)2661518-6 20734409 nnns volume:13 year:2023 number:1, p 14 https://doi.org/10.3390/cells13010014 kostenfrei https://doaj.org/article/d7f9b4d52a7645c69f1d564e94edeec5 kostenfrei https://www.mdpi.com/2073-4409/13/1/14 kostenfrei https://doaj.org/toc/2073-4409 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 13 2023 1, p 14 |
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10.3390/cells13010014 doi (DE-627)DOAJ097807907 (DE-599)DOAJd7f9b4d52a7645c69f1d564e94edeec5 DE-627 ger DE-627 rakwb eng QH573-671 Miao Chen verfasserin aut Brain Region Differences in α1- and α5-Subunit-Containing GABA<sub<A</sub< Receptor Proteomes Revealed with Affinity Purification and Blue Native PAGE Proteomics 2023 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier GABA<sub<A</sub< receptors are the major inhibitory receptors in the brain. They are hetero-pentamers with a composition of predominantly two α, two β, and one γ or δ subunit. Of the six α subunit genes, the α5 subunit displays a limited spatial expression pattern and is known to mediate both phasic and tonic inhibition. In this study, using immunoaffinity-based proteomics, we identified the α5 subunit containing receptor complexes in the hippocampus and olfactory bulb. The α1–α5 interaction was identified in both brain regions, albeit with significantly different stoichiometries. In line with this, reverse IPs using anti-α1 antibodies showed the α5–α1 co-occurrence and validated the quantitative difference. In addition, we showed that the association of Neuroligin 2 with α1-containing receptors was much higher in the olfactory bulb than in the hippocampus, which was confirmed using blue native gel electrophoresis and quantitative mass spectrometry. Finally, immunocytochemical staining revealed a co-localization of α1 and α5 subunits in the post-synaptic puncta in the hippocampus. GABA<sub<A</sub< receptor immunoprecipitation proteomics brain synapse immunocytochemistry Cytology Frank Koopmans verfasserin aut Miguel A. Gonzalez-Lozano verfasserin aut August B. Smit verfasserin aut Ka Wan Li verfasserin aut In Cells MDPI AG, 2012 13(2023), 1, p 14 (DE-627)718622081 (DE-600)2661518-6 20734409 nnns volume:13 year:2023 number:1, p 14 https://doi.org/10.3390/cells13010014 kostenfrei https://doaj.org/article/d7f9b4d52a7645c69f1d564e94edeec5 kostenfrei https://www.mdpi.com/2073-4409/13/1/14 kostenfrei https://doaj.org/toc/2073-4409 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 13 2023 1, p 14 |
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10.3390/cells13010014 doi (DE-627)DOAJ097807907 (DE-599)DOAJd7f9b4d52a7645c69f1d564e94edeec5 DE-627 ger DE-627 rakwb eng QH573-671 Miao Chen verfasserin aut Brain Region Differences in α1- and α5-Subunit-Containing GABA<sub<A</sub< Receptor Proteomes Revealed with Affinity Purification and Blue Native PAGE Proteomics 2023 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier GABA<sub<A</sub< receptors are the major inhibitory receptors in the brain. They are hetero-pentamers with a composition of predominantly two α, two β, and one γ or δ subunit. Of the six α subunit genes, the α5 subunit displays a limited spatial expression pattern and is known to mediate both phasic and tonic inhibition. In this study, using immunoaffinity-based proteomics, we identified the α5 subunit containing receptor complexes in the hippocampus and olfactory bulb. The α1–α5 interaction was identified in both brain regions, albeit with significantly different stoichiometries. In line with this, reverse IPs using anti-α1 antibodies showed the α5–α1 co-occurrence and validated the quantitative difference. In addition, we showed that the association of Neuroligin 2 with α1-containing receptors was much higher in the olfactory bulb than in the hippocampus, which was confirmed using blue native gel electrophoresis and quantitative mass spectrometry. Finally, immunocytochemical staining revealed a co-localization of α1 and α5 subunits in the post-synaptic puncta in the hippocampus. GABA<sub<A</sub< receptor immunoprecipitation proteomics brain synapse immunocytochemistry Cytology Frank Koopmans verfasserin aut Miguel A. Gonzalez-Lozano verfasserin aut August B. Smit verfasserin aut Ka Wan Li verfasserin aut In Cells MDPI AG, 2012 13(2023), 1, p 14 (DE-627)718622081 (DE-600)2661518-6 20734409 nnns volume:13 year:2023 number:1, p 14 https://doi.org/10.3390/cells13010014 kostenfrei https://doaj.org/article/d7f9b4d52a7645c69f1d564e94edeec5 kostenfrei https://www.mdpi.com/2073-4409/13/1/14 kostenfrei https://doaj.org/toc/2073-4409 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 13 2023 1, p 14 |
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Brain Region Differences in α1- and α5-Subunit-Containing GABA<sub<A</sub< Receptor Proteomes Revealed with Affinity Purification and Blue Native PAGE Proteomics |
abstract |
GABA<sub<A</sub< receptors are the major inhibitory receptors in the brain. They are hetero-pentamers with a composition of predominantly two α, two β, and one γ or δ subunit. Of the six α subunit genes, the α5 subunit displays a limited spatial expression pattern and is known to mediate both phasic and tonic inhibition. In this study, using immunoaffinity-based proteomics, we identified the α5 subunit containing receptor complexes in the hippocampus and olfactory bulb. The α1–α5 interaction was identified in both brain regions, albeit with significantly different stoichiometries. In line with this, reverse IPs using anti-α1 antibodies showed the α5–α1 co-occurrence and validated the quantitative difference. In addition, we showed that the association of Neuroligin 2 with α1-containing receptors was much higher in the olfactory bulb than in the hippocampus, which was confirmed using blue native gel electrophoresis and quantitative mass spectrometry. Finally, immunocytochemical staining revealed a co-localization of α1 and α5 subunits in the post-synaptic puncta in the hippocampus. |
abstractGer |
GABA<sub<A</sub< receptors are the major inhibitory receptors in the brain. They are hetero-pentamers with a composition of predominantly two α, two β, and one γ or δ subunit. Of the six α subunit genes, the α5 subunit displays a limited spatial expression pattern and is known to mediate both phasic and tonic inhibition. In this study, using immunoaffinity-based proteomics, we identified the α5 subunit containing receptor complexes in the hippocampus and olfactory bulb. The α1–α5 interaction was identified in both brain regions, albeit with significantly different stoichiometries. In line with this, reverse IPs using anti-α1 antibodies showed the α5–α1 co-occurrence and validated the quantitative difference. In addition, we showed that the association of Neuroligin 2 with α1-containing receptors was much higher in the olfactory bulb than in the hippocampus, which was confirmed using blue native gel electrophoresis and quantitative mass spectrometry. Finally, immunocytochemical staining revealed a co-localization of α1 and α5 subunits in the post-synaptic puncta in the hippocampus. |
abstract_unstemmed |
GABA<sub<A</sub< receptors are the major inhibitory receptors in the brain. They are hetero-pentamers with a composition of predominantly two α, two β, and one γ or δ subunit. Of the six α subunit genes, the α5 subunit displays a limited spatial expression pattern and is known to mediate both phasic and tonic inhibition. In this study, using immunoaffinity-based proteomics, we identified the α5 subunit containing receptor complexes in the hippocampus and olfactory bulb. The α1–α5 interaction was identified in both brain regions, albeit with significantly different stoichiometries. In line with this, reverse IPs using anti-α1 antibodies showed the α5–α1 co-occurrence and validated the quantitative difference. In addition, we showed that the association of Neuroligin 2 with α1-containing receptors was much higher in the olfactory bulb than in the hippocampus, which was confirmed using blue native gel electrophoresis and quantitative mass spectrometry. Finally, immunocytochemical staining revealed a co-localization of α1 and α5 subunits in the post-synaptic puncta in the hippocampus. |
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