Activation of Heteromeric G Protein-Gated Inward Rectifier K+ Channels Overexpressed by Adenovirus Gene Transfer Inhibits the Excitability of Hippocampal Neurons
G protein-gated inward rectifier K+ channel subunits 1-4 (GIRK1-4) have been cloned from neuronal and atrial tissue and function as heterotetramers. To examine the inhibition of neuronal excitation by GIRKs, we overexpressed GIRKs in cultured hippocampal neurons from 18 day rat embryos, which normal...
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| Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 1997-06, Vol.94 (13), p.7070-7075 |
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| container_title | Proceedings of the National Academy of Sciences - PNAS |
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| creator | Ehrengruber, Markus U. Doupnik, Craig A. Xu, Youfeng Garvey, Justine Jasek, Mark C. Lester, Henry A. Davidson, Norman |
| description | G protein-gated inward rectifier K+ channel subunits 1-4 (GIRK1-4) have been cloned from neuronal and atrial tissue and function as heterotetramers. To examine the inhibition of neuronal excitation by GIRKs, we overexpressed GIRKs in cultured hippocampal neurons from 18 day rat embryos, which normally lack or show low amounts of GIRK protein and currents. Adenoviral recombinants containing the cDNAs for GIRK1, GIRK2, GIRK4, and the serotonin 1A receptor were constructed. Typical GIRK currents could be activated by endogenous GABAB, serotonin 5-HT1A, and adenosine A1 receptors in neurons coinfected with GIRK1+2 or GIRK1+4. Under current clamp, GIRK activation increased the cell membrane conductance by 1- to 2-fold, hyperpolarized the cell by 11-14 mV, and inhibited action potential firing by increasing the threshold current for firing by 2- to 3-fold. These effects were not found in non- and mock-infected neurons, and were similar to the effects of muscarinic stimulation of native GIRK currents in atrial myocytes. Two inhibitory effects of GIRK activation, hyperpolarization and diminution of depolarizing pulses, were simulated from the experimental data. These inhibitory effects are physiologically important in the voltage range between the resting membrane potential and the potential where voltage-gated Na+ and K+ currents are activated; that is where GIRK currents are outward. |
| doi_str_mv | 10.1073/pnas.94.13.7070 |
| format | Article |
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To examine the inhibition of neuronal excitation by GIRKs, we overexpressed GIRKs in cultured hippocampal neurons from 18 day rat embryos, which normally lack or show low amounts of GIRK protein and currents. Adenoviral recombinants containing the cDNAs for GIRK1, GIRK2, GIRK4, and the serotonin 1A receptor were constructed. Typical GIRK currents could be activated by endogenous GABAB, serotonin 5-HT1A, and adenosine A1 receptors in neurons coinfected with GIRK1+2 or GIRK1+4. Under current clamp, GIRK activation increased the cell membrane conductance by 1- to 2-fold, hyperpolarized the cell by 11-14 mV, and inhibited action potential firing by increasing the threshold current for firing by 2- to 3-fold. These effects were not found in non- and mock-infected neurons, and were similar to the effects of muscarinic stimulation of native GIRK currents in atrial myocytes. Two inhibitory effects of GIRK activation, hyperpolarization and diminution of depolarizing pulses, were simulated from the experimental data. These inhibitory effects are physiologically important in the voltage range between the resting membrane potential and the potential where voltage-gated Na+ and K+ currents are activated; that is where GIRK currents are outward.</description><identifier>ISSN: 0027-8424</identifier><identifier>EISSN: 1091-6490</identifier><identifier>DOI: 10.1073/pnas.94.13.7070</identifier><identifier>PMID: 9192693</identifier><language>eng</language><publisher>United States: National Academy of Sciences of the United States of America</publisher><subject>Action Potentials ; Adenoviridae - genetics ; Adenoviruses ; Agonists ; Anatomy & physiology ; Animals ; Biological Sciences ; Cell membranes ; Cells, Cultured ; CHO Cells ; Cricetinae ; G Protein-Coupled Inwardly-Rectifying Potassium Channels ; Gene Expression ; Gene Transfer Techniques ; GTP-Binding Proteins - genetics ; GTP-Binding Proteins - metabolism ; Hippocampus ; Hippocampus - cytology ; Hippocampus - metabolism ; Ion Channel Gating ; Neurology ; Neurons ; Neurons - metabolism ; Potassium Channels - genetics ; Potassium Channels - metabolism ; Potassium Channels - physiology ; Potassium Channels, Inwardly Rectifying ; Pyramidal cells ; Rats ; Receptors ; Serotonin receptors ; Xenopus</subject><ispartof>Proceedings of the National Academy of Sciences - PNAS, 1997-06, Vol.94 (13), p.7070-7075</ispartof><rights>Copyright 1997 National Academy of Sciences</rights><rights>Copyright National Academy of Sciences Jun 24, 1997</rights><rights>Copyright © 1997, The National Academy of Sciences of the USA 1997</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c553t-9c9076c6da316bbceb3c4d2c208e2e259c6a5e1dc484c344da8e163672dbaacc3</citedby><cites>FETCH-LOGICAL-c553t-9c9076c6da316bbceb3c4d2c208e2e259c6a5e1dc484c344da8e163672dbaacc3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://www.pnas.org/content/94/13.cover.gif</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/42280$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/42280$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,314,723,776,780,799,881,27901,27902,53766,53768,57992,58225</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/9192693$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Ehrengruber, Markus U.</creatorcontrib><creatorcontrib>Doupnik, Craig A.</creatorcontrib><creatorcontrib>Xu, Youfeng</creatorcontrib><creatorcontrib>Garvey, Justine</creatorcontrib><creatorcontrib>Jasek, Mark C.</creatorcontrib><creatorcontrib>Lester, Henry A.</creatorcontrib><creatorcontrib>Davidson, Norman</creatorcontrib><title>Activation of Heteromeric G Protein-Gated Inward Rectifier K+ Channels Overexpressed by Adenovirus Gene Transfer Inhibits the Excitability of Hippocampal Neurons</title><title>Proceedings of the National Academy of Sciences - PNAS</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>G protein-gated inward rectifier K+ channel subunits 1-4 (GIRK1-4) have been cloned from neuronal and atrial tissue and function as heterotetramers. To examine the inhibition of neuronal excitation by GIRKs, we overexpressed GIRKs in cultured hippocampal neurons from 18 day rat embryos, which normally lack or show low amounts of GIRK protein and currents. Adenoviral recombinants containing the cDNAs for GIRK1, GIRK2, GIRK4, and the serotonin 1A receptor were constructed. Typical GIRK currents could be activated by endogenous GABAB, serotonin 5-HT1A, and adenosine A1 receptors in neurons coinfected with GIRK1+2 or GIRK1+4. Under current clamp, GIRK activation increased the cell membrane conductance by 1- to 2-fold, hyperpolarized the cell by 11-14 mV, and inhibited action potential firing by increasing the threshold current for firing by 2- to 3-fold. These effects were not found in non- and mock-infected neurons, and were similar to the effects of muscarinic stimulation of native GIRK currents in atrial myocytes. Two inhibitory effects of GIRK activation, hyperpolarization and diminution of depolarizing pulses, were simulated from the experimental data. 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To examine the inhibition of neuronal excitation by GIRKs, we overexpressed GIRKs in cultured hippocampal neurons from 18 day rat embryos, which normally lack or show low amounts of GIRK protein and currents. Adenoviral recombinants containing the cDNAs for GIRK1, GIRK2, GIRK4, and the serotonin 1A receptor were constructed. Typical GIRK currents could be activated by endogenous GABAB, serotonin 5-HT1A, and adenosine A1 receptors in neurons coinfected with GIRK1+2 or GIRK1+4. Under current clamp, GIRK activation increased the cell membrane conductance by 1- to 2-fold, hyperpolarized the cell by 11-14 mV, and inhibited action potential firing by increasing the threshold current for firing by 2- to 3-fold. These effects were not found in non- and mock-infected neurons, and were similar to the effects of muscarinic stimulation of native GIRK currents in atrial myocytes. Two inhibitory effects of GIRK activation, hyperpolarization and diminution of depolarizing pulses, were simulated from the experimental data. These inhibitory effects are physiologically important in the voltage range between the resting membrane potential and the potential where voltage-gated Na+ and K+ currents are activated; that is where GIRK currents are outward.</abstract><cop>United States</cop><pub>National Academy of Sciences of the United States of America</pub><pmid>9192693</pmid><doi>10.1073/pnas.94.13.7070</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Action Potentials Adenoviridae - genetics Adenoviruses Agonists Anatomy & physiology Animals Biological Sciences Cell membranes Cells, Cultured CHO Cells Cricetinae G Protein-Coupled Inwardly-Rectifying Potassium Channels Gene Expression Gene Transfer Techniques GTP-Binding Proteins - genetics GTP-Binding Proteins - metabolism Hippocampus Hippocampus - cytology Hippocampus - metabolism Ion Channel Gating Neurology Neurons Neurons - metabolism Potassium Channels - genetics Potassium Channels - metabolism Potassium Channels - physiology Potassium Channels, Inwardly Rectifying Pyramidal cells Rats Receptors Serotonin receptors Xenopus |
| title | Activation of Heteromeric G Protein-Gated Inward Rectifier K+ Channels Overexpressed by Adenovirus Gene Transfer Inhibits the Excitability of Hippocampal Neurons |
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