Activation by intracellular GDP, metabolic inhibition and pinacidil of a glibenclamide‐sensitive K‐channel in smooth muscle cells of rat mesenteric artery

1 Single‐channel recordings were made from cell‐attached and isolated patches, and whole‐cell currents zwere recorded under voltage clamp from single smooth muscle cells obtained by enzymic digestion of a small branch of the rat mesenteric artery. 2 In single voltage‐clamped cells 1 mm uridine dipho...

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Veröffentlicht in:British journal of pharmacology 1995-02, Vol.114 (3), p.662-672
Hauptverfasser: Zhang, H., Bolton, T.B.
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description 1 Single‐channel recordings were made from cell‐attached and isolated patches, and whole‐cell currents zwere recorded under voltage clamp from single smooth muscle cells obtained by enzymic digestion of a small branch of the rat mesenteric artery. 2 In single voltage‐clamped cells 1 mm uridine diphosphate (UDP) or guanidine diphosphate (GDP) added to the pipette solution, or pinacidil (100 μm) a K‐channel opener (KCO) applied in the bathing solution, evoked an outward current of up to 100pA which was blocked by glibenclamide (10 μm). In single cells from which recordings were made by the ‘perforated patch’ (nystatin pipette) technique, metabolic inhibition by 1 mm NaCN and 10 mm 2‐deoxy‐glucose also evoked a similar glibenclamide‐sensitive current. 3 Single K‐channel activity was observed in cell‐attached patches only infrequently unless the metabolism of the cell was inhibited, whereupon channel activity blocked by glibenclamide was seen; pinacidil applied to the cell evoked similar glibenclamide‐sensitive channel activity. If the patch was pulled off the cell to form an isolated inside‐out patch, similar glibenclamide‐sensitive single‐channel currents were observed in the presence of UDP and/or pinacidil to those seen in cell‐attached mode; channel conductance was 20 pS (60:130 K‐gradient) and openings showed no voltage‐dependence and noisy inward currents, typical of the nucleoside diphosphate (NDP) activated K‐channel (KNDP) seen previously in rabbit portal vein. 4 Formation of an isolated inside‐out patch into an ATP‐free solution did not increase the probability of channel opening which declined with time even when some single‐channel activity had occurred in the cell‐attached mode before detachment. However, application of 1 mm UDP or GDP, but not ATP, to inside‐out patches evoked single‐channel activity. Application of ATP‐free solution to isolated patches, previously exposed to ATP and in which channel activity had been seen, did not evoke channel activity. 5 It is concluded that small conductance K‐channels (KNDP) open in smooth muscle cells from this small artery in response to UDP or GDP acting from the inside, or pinacidil acting from the outside; the same channels open during inhibition of metabolism presumably mainly due to the rise in nucleoside diphosphates, but a fall in the ATP concentration on the inside of the channel did not by itself evoke channel activity. Failure to respond to a fall in ATP concentration upon formation of an inside
doi_str_mv 10.1111/j.1476-5381.1995.tb17190.x
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In single cells from which recordings were made by the ‘perforated patch’ (nystatin pipette) technique, metabolic inhibition by 1 mm NaCN and 10 mm 2‐deoxy‐glucose also evoked a similar glibenclamide‐sensitive current. 3 Single K‐channel activity was observed in cell‐attached patches only infrequently unless the metabolism of the cell was inhibited, whereupon channel activity blocked by glibenclamide was seen; pinacidil applied to the cell evoked similar glibenclamide‐sensitive channel activity. If the patch was pulled off the cell to form an isolated inside‐out patch, similar glibenclamide‐sensitive single‐channel currents were observed in the presence of UDP and/or pinacidil to those seen in cell‐attached mode; channel conductance was 20 pS (60:130 K‐gradient) and openings showed no voltage‐dependence and noisy inward currents, typical of the nucleoside diphosphate (NDP) activated K‐channel (KNDP) seen previously in rabbit portal vein. 4 Formation of an isolated inside‐out patch into an ATP‐free solution did not increase the probability of channel opening which declined with time even when some single‐channel activity had occurred in the cell‐attached mode before detachment. However, application of 1 mm UDP or GDP, but not ATP, to inside‐out patches evoked single‐channel activity. Application of ATP‐free solution to isolated patches, previously exposed to ATP and in which channel activity had been seen, did not evoke channel activity. 5 It is concluded that small conductance K‐channels (KNDP) open in smooth muscle cells from this small artery in response to UDP or GDP acting from the inside, or pinacidil acting from the outside; the same channels open during inhibition of metabolism presumably mainly due to the rise in nucleoside diphosphates, but a fall in the ATP concentration on the inside of the channel did not by itself evoke channel activity. Failure to respond to a fall in ATP concentration upon formation of an inside‐out patch could not be due to dephosphorylation of the channel because sometimes it had been active previously during cell‐attached recording. NDPs, instead of ATP, are more important regulators of KNDP channels. It is suggested that the KNDP is the main target K‐channel for KCOs.</description><identifier>ISSN: 0007-1188</identifier><identifier>EISSN: 1476-5381</identifier><identifier>DOI: 10.1111/j.1476-5381.1995.tb17190.x</identifier><identifier>PMID: 7735693</identifier><identifier>CODEN: BJPCBM</identifier><language>eng</language><publisher>Oxford, UK: Blackwell Publishing Ltd</publisher><subject>2‐deoxyglucose ; Animals ; Biological and medical sciences ; cyanide ; Deoxyglucose - pharmacology ; Electrophysiology ; GDP ; Glyburide - pharmacology ; Guanidines - pharmacology ; Guanosine Diphosphate - metabolism ; KNDP ; Male ; Medical sciences ; Mesenteric Arteries ; Metabolic inhibition ; Muscle ; Muscle Contraction - drug effects ; Muscle, Smooth, Vascular - drug effects ; Patch-Clamp Techniques ; Pharmacology. Drug treatments ; Pinacidil ; Potassium Channels - drug effects ; Rabbits ; Rats ; Rats, Wistar ; Sodium Cyanide - pharmacology ; uridine diphosphate ; Uridine Diphosphate - metabolism ; Vasodilator Agents - pharmacology</subject><ispartof>British journal of pharmacology, 1995-02, Vol.114 (3), p.662-672</ispartof><rights>1995 British Pharmacological Society</rights><rights>1995 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c5730-8f32ed4598aa7bf3ecb87343751dfe8aa47fe56bc87d0ce1357dccc588c2f9323</citedby><cites>FETCH-LOGICAL-c5730-8f32ed4598aa7bf3ecb87343751dfe8aa47fe56bc87d0ce1357dccc588c2f9323</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/PMC1510010/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC1510010/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,727,780,784,885,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&amp;idt=3411697$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/7735693$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Zhang, H.</creatorcontrib><creatorcontrib>Bolton, T.B.</creatorcontrib><title>Activation by intracellular GDP, metabolic inhibition and pinacidil of a glibenclamide‐sensitive K‐channel in smooth muscle cells of rat mesenteric artery</title><title>British journal of pharmacology</title><addtitle>Br J Pharmacol</addtitle><description>1 Single‐channel recordings were made from cell‐attached and isolated patches, and whole‐cell currents zwere recorded under voltage clamp from single smooth muscle cells obtained by enzymic digestion of a small branch of the rat mesenteric artery. 2 In single voltage‐clamped cells 1 mm uridine diphosphate (UDP) or guanidine diphosphate (GDP) added to the pipette solution, or pinacidil (100 μm) a K‐channel opener (KCO) applied in the bathing solution, evoked an outward current of up to 100pA which was blocked by glibenclamide (10 μm). In single cells from which recordings were made by the ‘perforated patch’ (nystatin pipette) technique, metabolic inhibition by 1 mm NaCN and 10 mm 2‐deoxy‐glucose also evoked a similar glibenclamide‐sensitive current. 3 Single K‐channel activity was observed in cell‐attached patches only infrequently unless the metabolism of the cell was inhibited, whereupon channel activity blocked by glibenclamide was seen; pinacidil applied to the cell evoked similar glibenclamide‐sensitive channel activity. If the patch was pulled off the cell to form an isolated inside‐out patch, similar glibenclamide‐sensitive single‐channel currents were observed in the presence of UDP and/or pinacidil to those seen in cell‐attached mode; channel conductance was 20 pS (60:130 K‐gradient) and openings showed no voltage‐dependence and noisy inward currents, typical of the nucleoside diphosphate (NDP) activated K‐channel (KNDP) seen previously in rabbit portal vein. 4 Formation of an isolated inside‐out patch into an ATP‐free solution did not increase the probability of channel opening which declined with time even when some single‐channel activity had occurred in the cell‐attached mode before detachment. However, application of 1 mm UDP or GDP, but not ATP, to inside‐out patches evoked single‐channel activity. Application of ATP‐free solution to isolated patches, previously exposed to ATP and in which channel activity had been seen, did not evoke channel activity. 5 It is concluded that small conductance K‐channels (KNDP) open in smooth muscle cells from this small artery in response to UDP or GDP acting from the inside, or pinacidil acting from the outside; the same channels open during inhibition of metabolism presumably mainly due to the rise in nucleoside diphosphates, but a fall in the ATP concentration on the inside of the channel did not by itself evoke channel activity. Failure to respond to a fall in ATP concentration upon formation of an inside‐out patch could not be due to dephosphorylation of the channel because sometimes it had been active previously during cell‐attached recording. NDPs, instead of ATP, are more important regulators of KNDP channels. It is suggested that the KNDP is the main target K‐channel for KCOs.</description><subject>2‐deoxyglucose</subject><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>cyanide</subject><subject>Deoxyglucose - pharmacology</subject><subject>Electrophysiology</subject><subject>GDP</subject><subject>Glyburide - pharmacology</subject><subject>Guanidines - pharmacology</subject><subject>Guanosine Diphosphate - metabolism</subject><subject>KNDP</subject><subject>Male</subject><subject>Medical sciences</subject><subject>Mesenteric Arteries</subject><subject>Metabolic inhibition</subject><subject>Muscle</subject><subject>Muscle Contraction - drug effects</subject><subject>Muscle, Smooth, Vascular - drug effects</subject><subject>Patch-Clamp Techniques</subject><subject>Pharmacology. Drug treatments</subject><subject>Pinacidil</subject><subject>Potassium Channels - drug effects</subject><subject>Rabbits</subject><subject>Rats</subject><subject>Rats, Wistar</subject><subject>Sodium Cyanide - pharmacology</subject><subject>uridine diphosphate</subject><subject>Uridine Diphosphate - metabolism</subject><subject>Vasodilator Agents - pharmacology</subject><issn>0007-1188</issn><issn>1476-5381</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1995</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqVUU1uEzEUthCohMIRkCzEkhnseBx7WKCWAi2iEl3A2nrzxtM48ngiexKaHUfgBByOk-BpogiWePNsf3-WP0JecFbyvF6vSl6pRSGF5iWva1mODVe8ZuXdAzI7Qg_JjDGmCs61fkyepLRiLINKnpATpYRc1GJGfp3j6LYwuiHQZkddGCOg9X7jIdLL9zevaG9HaAbvMINL17h7KoSWrl0AdK3zdOgo0FvvGhvQQ-9a-_vHz2RDyuStpZ_zCZcQgvXZg6Z-GMYl7TcJvaVTWJocIow5K6tGG3MYxDx3T8mjDnyyzw7zlHz7-OHrxVVx_eXy08X5dYFSCVboTsxtW8laA6imExYbrUQllORtZ_NlpTorFw1q1TK0XEjVIqLUGuddLebilLzd-643TW9btNM_eLOOroe4MwM48y8S3NLcDlvDJc-_yrLBm70BxiGlaLujljMzlWZWZmrGTM2YqTRzKM3cZfHzv9OP0kNLGX95wCEh-C5CQJeONFFxvqhVpp3tad-dt7v_eIB5d3N1vxV_AP0AvDc</recordid><startdate>199502</startdate><enddate>199502</enddate><creator>Zhang, H.</creator><creator>Bolton, T.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>5PM</scope></search><sort><creationdate>199502</creationdate><title>Activation by intracellular GDP, metabolic inhibition and pinacidil of a glibenclamide‐sensitive K‐channel in smooth muscle cells of rat mesenteric artery</title><author>Zhang, H. ; Bolton, T.B.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c5730-8f32ed4598aa7bf3ecb87343751dfe8aa47fe56bc87d0ce1357dccc588c2f9323</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1995</creationdate><topic>2‐deoxyglucose</topic><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>cyanide</topic><topic>Deoxyglucose - pharmacology</topic><topic>Electrophysiology</topic><topic>GDP</topic><topic>Glyburide - pharmacology</topic><topic>Guanidines - pharmacology</topic><topic>Guanosine Diphosphate - metabolism</topic><topic>KNDP</topic><topic>Male</topic><topic>Medical sciences</topic><topic>Mesenteric Arteries</topic><topic>Metabolic inhibition</topic><topic>Muscle</topic><topic>Muscle Contraction - drug effects</topic><topic>Muscle, Smooth, Vascular - drug effects</topic><topic>Patch-Clamp Techniques</topic><topic>Pharmacology. Drug treatments</topic><topic>Pinacidil</topic><topic>Potassium Channels - drug effects</topic><topic>Rabbits</topic><topic>Rats</topic><topic>Rats, Wistar</topic><topic>Sodium Cyanide - pharmacology</topic><topic>uridine diphosphate</topic><topic>Uridine Diphosphate - metabolism</topic><topic>Vasodilator Agents - pharmacology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Zhang, H.</creatorcontrib><creatorcontrib>Bolton, T.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>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>Zhang, H.</au><au>Bolton, T.B.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Activation by intracellular GDP, metabolic inhibition and pinacidil of a glibenclamide‐sensitive K‐channel in smooth muscle cells of rat mesenteric artery</atitle><jtitle>British journal of pharmacology</jtitle><addtitle>Br J Pharmacol</addtitle><date>1995-02</date><risdate>1995</risdate><volume>114</volume><issue>3</issue><spage>662</spage><epage>672</epage><pages>662-672</pages><issn>0007-1188</issn><eissn>1476-5381</eissn><coden>BJPCBM</coden><abstract>1 Single‐channel recordings were made from cell‐attached and isolated patches, and whole‐cell currents zwere recorded under voltage clamp from single smooth muscle cells obtained by enzymic digestion of a small branch of the rat mesenteric artery. 2 In single voltage‐clamped cells 1 mm uridine diphosphate (UDP) or guanidine diphosphate (GDP) added to the pipette solution, or pinacidil (100 μm) a K‐channel opener (KCO) applied in the bathing solution, evoked an outward current of up to 100pA which was blocked by glibenclamide (10 μm). In single cells from which recordings were made by the ‘perforated patch’ (nystatin pipette) technique, metabolic inhibition by 1 mm NaCN and 10 mm 2‐deoxy‐glucose also evoked a similar glibenclamide‐sensitive current. 3 Single K‐channel activity was observed in cell‐attached patches only infrequently unless the metabolism of the cell was inhibited, whereupon channel activity blocked by glibenclamide was seen; pinacidil applied to the cell evoked similar glibenclamide‐sensitive channel activity. If the patch was pulled off the cell to form an isolated inside‐out patch, similar glibenclamide‐sensitive single‐channel currents were observed in the presence of UDP and/or pinacidil to those seen in cell‐attached mode; channel conductance was 20 pS (60:130 K‐gradient) and openings showed no voltage‐dependence and noisy inward currents, typical of the nucleoside diphosphate (NDP) activated K‐channel (KNDP) seen previously in rabbit portal vein. 4 Formation of an isolated inside‐out patch into an ATP‐free solution did not increase the probability of channel opening which declined with time even when some single‐channel activity had occurred in the cell‐attached mode before detachment. However, application of 1 mm UDP or GDP, but not ATP, to inside‐out patches evoked single‐channel activity. Application of ATP‐free solution to isolated patches, previously exposed to ATP and in which channel activity had been seen, did not evoke channel activity. 5 It is concluded that small conductance K‐channels (KNDP) open in smooth muscle cells from this small artery in response to UDP or GDP acting from the inside, or pinacidil acting from the outside; the same channels open during inhibition of metabolism presumably mainly due to the rise in nucleoside diphosphates, but a fall in the ATP concentration on the inside of the channel did not by itself evoke channel activity. Failure to respond to a fall in ATP concentration upon formation of an inside‐out patch could not be due to dephosphorylation of the channel because sometimes it had been active previously during cell‐attached recording. NDPs, instead of ATP, are more important regulators of KNDP channels. It is suggested that the KNDP is the main target K‐channel for KCOs.</abstract><cop>Oxford, UK</cop><pub>Blackwell Publishing Ltd</pub><pmid>7735693</pmid><doi>10.1111/j.1476-5381.1995.tb17190.x</doi><tpages>11</tpages><oa>free_for_read</oa></addata></record>
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source MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central; Alma/SFX Local Collection
subjects 2‐deoxyglucose
Animals
Biological and medical sciences
cyanide
Deoxyglucose - pharmacology
Electrophysiology
GDP
Glyburide - pharmacology
Guanidines - pharmacology
Guanosine Diphosphate - metabolism
KNDP
Male
Medical sciences
Mesenteric Arteries
Metabolic inhibition
Muscle
Muscle Contraction - drug effects
Muscle, Smooth, Vascular - drug effects
Patch-Clamp Techniques
Pharmacology. Drug treatments
Pinacidil
Potassium Channels - drug effects
Rabbits
Rats
Rats, Wistar
Sodium Cyanide - pharmacology
uridine diphosphate
Uridine Diphosphate - metabolism
Vasodilator Agents - pharmacology
title Activation by intracellular GDP, metabolic inhibition and pinacidil of a glibenclamide‐sensitive K‐channel in smooth muscle cells of rat mesenteric artery
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