Single channel analysis of the blocking actions of BIDN and fipronil on a Drosophila melanogaster GABA receptor (RDL) stably expressed in a Drosophila cell line
Single channel recordings were obtained from a Drosophila S2 cell line stably expressing the wild‐type RDLac Drosophila melanogaster homomer‐forming ionotropic GABA receptor subunit, a product of the resistance to dieldrin gene, Rdl. GABA (50 μM) was applied by pressure ejection to outside‐out patch...
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| description | Single channel recordings were obtained from a Drosophila S2 cell line stably expressing the wild‐type RDLac Drosophila melanogaster homomer‐forming ionotropic GABA receptor subunit, a product of the resistance to dieldrin gene, Rdl. GABA (50 μM) was applied by pressure ejection to outside‐out patches from S2‐RDL cells at a holding potential of −60 mV. The resulting inward current was completely blocked by 100 μM picrotoxin (PTX). The unitary current‐voltage relationship was linear at negative potentials but showed slight inward rectification at potentials more positive than 0 mV. The reversal potential of the current (EGABA=−1.4 mV) was close to the calculated chloride equilibrium potential.
The single channel conductance elicited by GABA was 36 pS. A 71 pS conductance channel was also observed when the duration of the pulse, used to eject GABA, was longer than 80 ms. The mean open time distribution of the unitary events was fitted best by two exponential functions suggesting two open channel states.
When either 1 μM fipronil or 1 μM BIDN was present in the external saline, the GABA‐gated channels were completely blocked. When BIDN or fipronil was applied at a concentration close to the IC50 value for suppression of open probability (281 nM, BIDN; 240 nM, fipronil), the duration of channel openings was shortened. In addition, the blocking action of BIDN resulted in the appearance of a novel channel conductance (17 pS).
The effects of co‐application of BIDN and fipronil were examined. Co‐application of BIDN (300 nM) with various concentrations (100–1000 nM) of fipronil resulted in an additional BIDN‐induced dose‐dependent reduction of the maximum Po value.
Thus both BIDN and fipronil shorten the duration of wild‐type RDLac GABA receptor channel openings but appear to act at distinct sites.
British Journal of Pharmacology (2000) 130, 1833–1842; doi:10.1038/sj.bjp.0703507 |
| doi_str_mv | 10.1038/sj.bjp.0703507 |
| format | Article |
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The single channel conductance elicited by GABA was 36 pS. A 71 pS conductance channel was also observed when the duration of the pulse, used to eject GABA, was longer than 80 ms. The mean open time distribution of the unitary events was fitted best by two exponential functions suggesting two open channel states.
When either 1 μM fipronil or 1 μM BIDN was present in the external saline, the GABA‐gated channels were completely blocked. When BIDN or fipronil was applied at a concentration close to the IC50 value for suppression of open probability (281 nM, BIDN; 240 nM, fipronil), the duration of channel openings was shortened. In addition, the blocking action of BIDN resulted in the appearance of a novel channel conductance (17 pS).
The effects of co‐application of BIDN and fipronil were examined. Co‐application of BIDN (300 nM) with various concentrations (100–1000 nM) of fipronil resulted in an additional BIDN‐induced dose‐dependent reduction of the maximum Po value.
Thus both BIDN and fipronil shorten the duration of wild‐type RDLac GABA receptor channel openings but appear to act at distinct sites.
British Journal of Pharmacology (2000) 130, 1833–1842; doi:10.1038/sj.bjp.0703507</description><identifier>ISSN: 0007-1188</identifier><identifier>EISSN: 1476-5381</identifier><identifier>DOI: 10.1038/sj.bjp.0703507</identifier><identifier>PMID: 10952672</identifier><identifier>CODEN: BJPCBM</identifier><language>eng</language><publisher>Oxford, UK: Blackwell Publishing Ltd</publisher><subject>Animals ; BIDN ; Biological and medical sciences ; Bridged Bicyclo Compounds - pharmacology ; Cell Line ; Channel opening ; Chemical control ; Chloride ; Conductance ; Control ; Dieldrin ; Dose-Response Relationship, Drug ; Drosophila melanogaster ; Drosophila melanogaster - cytology ; Drosophila melanogaster - drug effects ; Drosophila melanogaster - physiology ; Drosophila Proteins ; fipronil ; Fundamental and applied biological sciences. Psychology ; GABA Antagonists - pharmacology ; GABA receptor ; gamma -Aminobutyric acid ; gamma -Aminobutyric acid receptors ; gamma-Aminobutyric Acid - pharmacology ; Ion Channels - drug effects ; Membrane Potentials - drug effects ; Nitriles - pharmacology ; Patch-Clamp Techniques ; Pharmacology ; Phytopathology. Animal pests. Plant and forest protection ; picrotoxin ; Picrotoxin - pharmacology ; Pressure ; Protozoa. Invertebrates ; Pyrazoles - pharmacology ; Receptors, GABA-A - drug effects ; single channel ; stable cell line ; Time Factors</subject><ispartof>British journal of pharmacology, 2000-08, Vol.130 (8), p.1833-1842</ispartof><rights>2000 British Pharmacological Society</rights><rights>2000 INIST-CNRS</rights><rights>Copyright Nature Publishing Group Aug 2000</rights><rights>Copyright 2000, Nature Publishing Group 2000 Nature Publishing Group</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c6162-a101ec02aeac763fcc47ddadfb4a01609bbff2acfdaf05333e4c3cc9259a5a883</citedby><cites>FETCH-LOGICAL-c6162-a101ec02aeac763fcc47ddadfb4a01609bbff2acfdaf05333e4c3cc9259a5a883</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC1572267/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC1572267/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,727,780,784,885,1417,1433,27924,27925,45574,45575,46409,46833,53791,53793</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=1484140$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/10952672$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Grolleau, Françoise</creatorcontrib><creatorcontrib>Sattelle, David B</creatorcontrib><title>Single channel analysis of the blocking actions of BIDN and fipronil on a Drosophila melanogaster GABA receptor (RDL) stably expressed in a Drosophila cell line</title><title>British journal of pharmacology</title><addtitle>Br J Pharmacol</addtitle><description>Single channel recordings were obtained from a Drosophila S2 cell line stably expressing the wild‐type RDLac Drosophila melanogaster homomer‐forming ionotropic GABA receptor subunit, a product of the resistance to dieldrin gene, Rdl. GABA (50 μM) was applied by pressure ejection to outside‐out patches from S2‐RDL cells at a holding potential of −60 mV. The resulting inward current was completely blocked by 100 μM picrotoxin (PTX). The unitary current‐voltage relationship was linear at negative potentials but showed slight inward rectification at potentials more positive than 0 mV. The reversal potential of the current (EGABA=−1.4 mV) was close to the calculated chloride equilibrium potential.
The single channel conductance elicited by GABA was 36 pS. A 71 pS conductance channel was also observed when the duration of the pulse, used to eject GABA, was longer than 80 ms. The mean open time distribution of the unitary events was fitted best by two exponential functions suggesting two open channel states.
When either 1 μM fipronil or 1 μM BIDN was present in the external saline, the GABA‐gated channels were completely blocked. When BIDN or fipronil was applied at a concentration close to the IC50 value for suppression of open probability (281 nM, BIDN; 240 nM, fipronil), the duration of channel openings was shortened. In addition, the blocking action of BIDN resulted in the appearance of a novel channel conductance (17 pS).
The effects of co‐application of BIDN and fipronil were examined. Co‐application of BIDN (300 nM) with various concentrations (100–1000 nM) of fipronil resulted in an additional BIDN‐induced dose‐dependent reduction of the maximum Po value.
Thus both BIDN and fipronil shorten the duration of wild‐type RDLac GABA receptor channel openings but appear to act at distinct sites.
British Journal of Pharmacology (2000) 130, 1833–1842; doi:10.1038/sj.bjp.0703507</description><subject>Animals</subject><subject>BIDN</subject><subject>Biological and medical sciences</subject><subject>Bridged Bicyclo Compounds - pharmacology</subject><subject>Cell Line</subject><subject>Channel opening</subject><subject>Chemical control</subject><subject>Chloride</subject><subject>Conductance</subject><subject>Control</subject><subject>Dieldrin</subject><subject>Dose-Response Relationship, Drug</subject><subject>Drosophila melanogaster</subject><subject>Drosophila melanogaster - cytology</subject><subject>Drosophila melanogaster - drug effects</subject><subject>Drosophila melanogaster - physiology</subject><subject>Drosophila Proteins</subject><subject>fipronil</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>GABA Antagonists - pharmacology</subject><subject>GABA receptor</subject><subject>gamma -Aminobutyric acid</subject><subject>gamma -Aminobutyric acid receptors</subject><subject>gamma-Aminobutyric Acid - pharmacology</subject><subject>Ion Channels - drug effects</subject><subject>Membrane Potentials - drug effects</subject><subject>Nitriles - pharmacology</subject><subject>Patch-Clamp Techniques</subject><subject>Pharmacology</subject><subject>Phytopathology. Animal pests. Plant and forest protection</subject><subject>picrotoxin</subject><subject>Picrotoxin - pharmacology</subject><subject>Pressure</subject><subject>Protozoa. Invertebrates</subject><subject>Pyrazoles - pharmacology</subject><subject>Receptors, GABA-A - drug effects</subject><subject>single channel</subject><subject>stable cell line</subject><subject>Time Factors</subject><issn>0007-1188</issn><issn>1476-5381</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNqFkk2P0zAQhiMEYsvClSOyEELLIcWOkzi9ILVb2F2pAsTH2Zo449bFtYOdAv03_FTcbQXLHuA0kufxO19vlj1mdMwob17G9bhd92MqKK-ouJONWCnqvOINu5uNKKUiZ6xpTrIHMa4pTUlR3c9OGJ1URS2KUfbzo3FLi0StwDm0BBzYXTSReE2GFZLWevUlIQTUYLy7fp9dzd8msCPa9ME7Y4l3BMg8-Oj7lbFANmjB-SXEAQO5mM6mJKDCfvCBnH2YL16QOEBrdwR_9AFjxI6YWwoKrSXWOHyY3dNgIz46xtPs85vXn84v88W7i6vz6SJXNauLHBhlqGgBCErUXCtViq6DTrclUFbTSdtqXYDSHWhacc6xVFypSVFNoIKm4afZq4Nuv2032Cl0QwAr-2A2EHbSg5F_Z5xZyaX_JlklirTLJPD8KBD81y3GQW5M3I8BDv02SiZq1pTX4Nm_QVo0VNRltW_q6S107bchnSjKggmWjlnUCRofIJXWFwPq300zKvcmkXEtk0nk0STpw5Obo97AD65IwLMjAFGB1QGcMvEPVzYlK2nC-AH7bizu_lNVzt5fsmpS8F9e_9hS</recordid><startdate>200008</startdate><enddate>200008</enddate><creator>Grolleau, Françoise</creator><creator>Sattelle, David B</creator><general>Blackwell Publishing Ltd</general><general>Nature Publishing</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7QP</scope><scope>7RV</scope><scope>7TK</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7P</scope><scope>NAPCQ</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7SS</scope><scope>5PM</scope></search><sort><creationdate>200008</creationdate><title>Single channel analysis of the blocking actions of BIDN and fipronil on a Drosophila melanogaster GABA receptor (RDL) stably expressed in a Drosophila cell line</title><author>Grolleau, Françoise ; Sattelle, David B</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c6162-a101ec02aeac763fcc47ddadfb4a01609bbff2acfdaf05333e4c3cc9259a5a883</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2000</creationdate><topic>Animals</topic><topic>BIDN</topic><topic>Biological and medical sciences</topic><topic>Bridged Bicyclo Compounds - pharmacology</topic><topic>Cell Line</topic><topic>Channel opening</topic><topic>Chemical control</topic><topic>Chloride</topic><topic>Conductance</topic><topic>Control</topic><topic>Dieldrin</topic><topic>Dose-Response Relationship, Drug</topic><topic>Drosophila melanogaster</topic><topic>Drosophila melanogaster - cytology</topic><topic>Drosophila melanogaster - drug effects</topic><topic>Drosophila melanogaster - physiology</topic><topic>Drosophila Proteins</topic><topic>fipronil</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>GABA Antagonists - pharmacology</topic><topic>GABA receptor</topic><topic>gamma -Aminobutyric acid</topic><topic>gamma -Aminobutyric acid receptors</topic><topic>gamma-Aminobutyric Acid - pharmacology</topic><topic>Ion Channels - drug effects</topic><topic>Membrane Potentials - drug effects</topic><topic>Nitriles - pharmacology</topic><topic>Patch-Clamp Techniques</topic><topic>Pharmacology</topic><topic>Phytopathology. Animal pests. Plant and forest protection</topic><topic>picrotoxin</topic><topic>Picrotoxin - pharmacology</topic><topic>Pressure</topic><topic>Protozoa. Invertebrates</topic><topic>Pyrazoles - pharmacology</topic><topic>Receptors, GABA-A - drug effects</topic><topic>single channel</topic><topic>stable cell line</topic><topic>Time Factors</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Grolleau, Françoise</creatorcontrib><creatorcontrib>Sattelle, David B</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Nursing & Allied Health Database</collection><collection>Neurosciences Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection (ProQuest)</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Biological Science Database</collection><collection>Nursing & Allied Health Premium</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>Entomology Abstracts (Full archive)</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>British journal of pharmacology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Grolleau, Françoise</au><au>Sattelle, David B</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Single channel analysis of the blocking actions of BIDN and fipronil on a Drosophila melanogaster GABA receptor (RDL) stably expressed in a Drosophila cell line</atitle><jtitle>British journal of pharmacology</jtitle><addtitle>Br J Pharmacol</addtitle><date>2000-08</date><risdate>2000</risdate><volume>130</volume><issue>8</issue><spage>1833</spage><epage>1842</epage><pages>1833-1842</pages><issn>0007-1188</issn><eissn>1476-5381</eissn><coden>BJPCBM</coden><abstract>Single channel recordings were obtained from a Drosophila S2 cell line stably expressing the wild‐type RDLac Drosophila melanogaster homomer‐forming ionotropic GABA receptor subunit, a product of the resistance to dieldrin gene, Rdl. GABA (50 μM) was applied by pressure ejection to outside‐out patches from S2‐RDL cells at a holding potential of −60 mV. The resulting inward current was completely blocked by 100 μM picrotoxin (PTX). The unitary current‐voltage relationship was linear at negative potentials but showed slight inward rectification at potentials more positive than 0 mV. The reversal potential of the current (EGABA=−1.4 mV) was close to the calculated chloride equilibrium potential.
The single channel conductance elicited by GABA was 36 pS. A 71 pS conductance channel was also observed when the duration of the pulse, used to eject GABA, was longer than 80 ms. The mean open time distribution of the unitary events was fitted best by two exponential functions suggesting two open channel states.
When either 1 μM fipronil or 1 μM BIDN was present in the external saline, the GABA‐gated channels were completely blocked. When BIDN or fipronil was applied at a concentration close to the IC50 value for suppression of open probability (281 nM, BIDN; 240 nM, fipronil), the duration of channel openings was shortened. In addition, the blocking action of BIDN resulted in the appearance of a novel channel conductance (17 pS).
The effects of co‐application of BIDN and fipronil were examined. Co‐application of BIDN (300 nM) with various concentrations (100–1000 nM) of fipronil resulted in an additional BIDN‐induced dose‐dependent reduction of the maximum Po value.
Thus both BIDN and fipronil shorten the duration of wild‐type RDLac GABA receptor channel openings but appear to act at distinct sites.
British Journal of Pharmacology (2000) 130, 1833–1842; doi:10.1038/sj.bjp.0703507</abstract><cop>Oxford, UK</cop><pub>Blackwell Publishing Ltd</pub><pmid>10952672</pmid><doi>10.1038/sj.bjp.0703507</doi><tpages>10</tpages><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; Wiley Free Content; EZB-FREE-00999 freely available EZB journals; Wiley Online Library All Journals; PubMed Central; Alma/SFX Local Collection |
| subjects | Animals BIDN Biological and medical sciences Bridged Bicyclo Compounds - pharmacology Cell Line Channel opening Chemical control Chloride Conductance Control Dieldrin Dose-Response Relationship, Drug Drosophila melanogaster Drosophila melanogaster - cytology Drosophila melanogaster - drug effects Drosophila melanogaster - physiology Drosophila Proteins fipronil Fundamental and applied biological sciences. Psychology GABA Antagonists - pharmacology GABA receptor gamma -Aminobutyric acid gamma -Aminobutyric acid receptors gamma-Aminobutyric Acid - pharmacology Ion Channels - drug effects Membrane Potentials - drug effects Nitriles - pharmacology Patch-Clamp Techniques Pharmacology Phytopathology. Animal pests. Plant and forest protection picrotoxin Picrotoxin - pharmacology Pressure Protozoa. Invertebrates Pyrazoles - pharmacology Receptors, GABA-A - drug effects single channel stable cell line Time Factors |
| title | Single channel analysis of the blocking actions of BIDN and fipronil on a Drosophila melanogaster GABA receptor (RDL) stably expressed in a Drosophila cell line |
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