A quantitative investigation into the dependence of Ca2+ mobilisation on changes in inositol 1,4,5‐trisphosphate levels in the stimulated neutrophil

1 The coupling of N‐formyl‐methionyl‐leucyl‐phenylalanine (fMet‐Leu‐Phe) receptor stimulation to Ca2+ mobilisation has been investigated in the human neutrophil by measuring the concentration‐effect curves for inositol 1,4,5‐trisphosphate (IP3) formation and Ca2+ mobilisation. 2 fMet‐Leu‐Phe‐depende...

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Veröffentlicht in:British journal of pharmacology 1991-06, Vol.103 (2), p.1592-1596
Hauptverfasser: Thompson, Neil T., Bonser, Robert W., Tateson, James E., Spacey, Graham D., Randall, Roger W., Hodson, Harold F., Garland, Lawrence G.
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container_end_page 1596
container_issue 2
container_start_page 1592
container_title British journal of pharmacology
container_volume 103
creator Thompson, Neil T.
Bonser, Robert W.
Tateson, James E.
Spacey, Graham D.
Randall, Roger W.
Hodson, Harold F.
Garland, Lawrence G.
description 1 The coupling of N‐formyl‐methionyl‐leucyl‐phenylalanine (fMet‐Leu‐Phe) receptor stimulation to Ca2+ mobilisation has been investigated in the human neutrophil by measuring the concentration‐effect curves for inositol 1,4,5‐trisphosphate (IP3) formation and Ca2+ mobilisation. 2 fMet‐Leu‐Phe‐dependent mobilisation of intracellular Ca2+ has been monitored in fluo‐3‐loaded human neutrophils by measuring increases in the cytoplasmic free Ca2+ concentration ([Ca2+]i) in the presence of extracellular EGTA. Fluo‐3 was used in preference to fura‐2 because it was found to be more sensitive to the high Ca2+ levels seen in stimulated neutrophils. 3 fMet‐Leu‐Phe induced a rapid mobilisation of intracellular Ca2+ (EC50 = 2.9 ± 0.1 nm) and increased [Ca2+]i to a maximum of 1286 ± 184 nm. 4 The amount of IP3 in fMet‐Leu‐Phe‐stimulated neutrophils was determined by competition with [3H]‐IP3 for a specific IP3 binding protein isolated from bovine adrenocortical microsomes. Basal IP3 levels of 13.3 ± 2.0 pmol per 107 cells were increased nearly 4 fold by maximally effective concentrations of fMet‐Leu‐Phe. 5 The EC50 for the IP3 response (95 ± 18 nm) was much higher than that for mobilisation of intracellular Ca2+, such that only a doubling in the concentration of IP3 was required to fully mobilise intracellular Ca2+. 6 As a result of this relationship IP3 production was more sensitive than Ca2+ mobilisation to inhibition by demethoxyviridin, an inhibitor of phospholipase activation.
doi_str_mv 10.1111/j.1476-5381.1991.tb09832.x
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Fluo‐3 was used in preference to fura‐2 because it was found to be more sensitive to the high Ca2+ levels seen in stimulated neutrophils. 3 fMet‐Leu‐Phe induced a rapid mobilisation of intracellular Ca2+ (EC50 = 2.9 ± 0.1 nm) and increased [Ca2+]i to a maximum of 1286 ± 184 nm. 4 The amount of IP3 in fMet‐Leu‐Phe‐stimulated neutrophils was determined by competition with [3H]‐IP3 for a specific IP3 binding protein isolated from bovine adrenocortical microsomes. Basal IP3 levels of 13.3 ± 2.0 pmol per 107 cells were increased nearly 4 fold by maximally effective concentrations of fMet‐Leu‐Phe. 5 The EC50 for the IP3 response (95 ± 18 nm) was much higher than that for mobilisation of intracellular Ca2+, such that only a doubling in the concentration of IP3 was required to fully mobilise intracellular Ca2+. 6 As a result of this relationship IP3 production was more sensitive than Ca2+ mobilisation to inhibition by demethoxyviridin, an inhibitor of phospholipase activation.</description><identifier>ISSN: 0007-1188</identifier><identifier>EISSN: 1476-5381</identifier><identifier>DOI: 10.1111/j.1476-5381.1991.tb09832.x</identifier><identifier>PMID: 1884113</identifier><identifier>CODEN: BJPCBM</identifier><language>eng</language><publisher>Oxford, UK: Blackwell Publishing Ltd</publisher><subject>Aniline Compounds ; Biological and medical sciences ; Calcium - metabolism ; demethoxyvirdin ; Fluorescence ; Fundamental and applied biological sciences. Psychology ; Fundamental immunology ; Fura-2 ; Humans ; Immunobiology ; In Vitro Techniques ; Inositol 1,4,5-Trisphosphate - metabolism ; Myeloid cells: ontogeny, maturation, markers, receptors ; N-Formylmethionine Leucyl-Phenylalanine - pharmacology ; Neutrophils - drug effects ; Neutrophils - metabolism ; Phosphodiesterase Inhibitors - pharmacology ; phospholipase C ; phospholipase D ; phospholipase inhibitor ; Polynuclears ; Receptors, Formyl Peptide ; Receptors, Immunologic - drug effects ; Receptors, Immunologic - metabolism ; Signal transduction ; Signal Transduction - drug effects ; Xanthenes</subject><ispartof>British journal of pharmacology, 1991-06, Vol.103 (2), p.1592-1596</ispartof><rights>1991 British Pharmacological Society</rights><rights>1991 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC1908335/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC1908335/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,727,780,784,885,27922,27923,53789,53791</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&amp;idt=19669631$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1884113$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Thompson, Neil T.</creatorcontrib><creatorcontrib>Bonser, Robert W.</creatorcontrib><creatorcontrib>Tateson, James E.</creatorcontrib><creatorcontrib>Spacey, Graham D.</creatorcontrib><creatorcontrib>Randall, Roger W.</creatorcontrib><creatorcontrib>Hodson, Harold F.</creatorcontrib><creatorcontrib>Garland, Lawrence G.</creatorcontrib><title>A quantitative investigation into the dependence of Ca2+ mobilisation on changes in inositol 1,4,5‐trisphosphate levels in the stimulated neutrophil</title><title>British journal of pharmacology</title><addtitle>Br J Pharmacol</addtitle><description>1 The coupling of N‐formyl‐methionyl‐leucyl‐phenylalanine (fMet‐Leu‐Phe) receptor stimulation to Ca2+ mobilisation has been investigated in the human neutrophil by measuring the concentration‐effect curves for inositol 1,4,5‐trisphosphate (IP3) formation and Ca2+ mobilisation. 2 fMet‐Leu‐Phe‐dependent mobilisation of intracellular Ca2+ has been monitored in fluo‐3‐loaded human neutrophils by measuring increases in the cytoplasmic free Ca2+ concentration ([Ca2+]i) in the presence of extracellular EGTA. Fluo‐3 was used in preference to fura‐2 because it was found to be more sensitive to the high Ca2+ levels seen in stimulated neutrophils. 3 fMet‐Leu‐Phe induced a rapid mobilisation of intracellular Ca2+ (EC50 = 2.9 ± 0.1 nm) and increased [Ca2+]i to a maximum of 1286 ± 184 nm. 4 The amount of IP3 in fMet‐Leu‐Phe‐stimulated neutrophils was determined by competition with [3H]‐IP3 for a specific IP3 binding protein isolated from bovine adrenocortical microsomes. Basal IP3 levels of 13.3 ± 2.0 pmol per 107 cells were increased nearly 4 fold by maximally effective concentrations of fMet‐Leu‐Phe. 5 The EC50 for the IP3 response (95 ± 18 nm) was much higher than that for mobilisation of intracellular Ca2+, such that only a doubling in the concentration of IP3 was required to fully mobilise intracellular Ca2+. 6 As a result of this relationship IP3 production was more sensitive than Ca2+ mobilisation to inhibition by demethoxyviridin, an inhibitor of phospholipase activation.</description><subject>Aniline Compounds</subject><subject>Biological and medical sciences</subject><subject>Calcium - metabolism</subject><subject>demethoxyvirdin</subject><subject>Fluorescence</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Fundamental immunology</subject><subject>Fura-2</subject><subject>Humans</subject><subject>Immunobiology</subject><subject>In Vitro Techniques</subject><subject>Inositol 1,4,5-Trisphosphate - metabolism</subject><subject>Myeloid cells: ontogeny, maturation, markers, receptors</subject><subject>N-Formylmethionine Leucyl-Phenylalanine - pharmacology</subject><subject>Neutrophils - drug effects</subject><subject>Neutrophils - metabolism</subject><subject>Phosphodiesterase Inhibitors - pharmacology</subject><subject>phospholipase C</subject><subject>phospholipase D</subject><subject>phospholipase inhibitor</subject><subject>Polynuclears</subject><subject>Receptors, Formyl Peptide</subject><subject>Receptors, Immunologic - drug effects</subject><subject>Receptors, Immunologic - metabolism</subject><subject>Signal transduction</subject><subject>Signal Transduction - drug effects</subject><subject>Xanthenes</subject><issn>0007-1188</issn><issn>1476-5381</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1991</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpVUsFu1DAQtRCoLIVPQLKQ4EITPHEcxxdQWQFFqgQHOFvexLvxyrHT2FnaG5_AiQ_kS3C6UQvWWPbMe35jewahF0BySOPNPoeSVxmjNeQgBORxQ0RNi_z6AVrdQQ_RihDCM4C6foyehLAnJIGcnaCTFCoB6Ar9PsdXk3LRRBXNQWPjDjpEs0ued8mLHsdO41YP2rXaNRr7LV6r4jXu_cZYE47EZE2n3E6HdCaZDyZ6i-GsPGN_fv6KowlD59NUUWOrD9reEmfplK2fbIq32Okpjn7ojH2KHm2VDfrZsp6i7x8_fFtfZJdfPn1en19mA6VQZCCA1aIoNkrUqmhI2xIBhSJQM1Y3XKf3tgkrm4pTYNAKUbKKc8IrUacPA3qK3h51h2nT67bRLo7KymE0vRpvpFdG_o8408mdP0gQpKaUJYFXi8Dor6b0dbI3odHWKqf9FCQvCGes4In4_N9MdymWSiT85YKr0Ci7HZVrTLiniaoS1e2N3x15P4zVN_c4kXNryL2c6y_n-su5NeTSGvJavv96Me_oX-T8sJY</recordid><startdate>199106</startdate><enddate>199106</enddate><creator>Thompson, Neil T.</creator><creator>Bonser, Robert W.</creator><creator>Tateson, James E.</creator><creator>Spacey, Graham D.</creator><creator>Randall, Roger W.</creator><creator>Hodson, Harold F.</creator><creator>Garland, Lawrence G.</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>7X8</scope><scope>5PM</scope></search><sort><creationdate>199106</creationdate><title>A quantitative investigation into the dependence of Ca2+ mobilisation on changes in inositol 1,4,5‐trisphosphate levels in the stimulated neutrophil</title><author>Thompson, Neil T. ; Bonser, Robert W. ; Tateson, James E. ; Spacey, Graham D. ; Randall, Roger W. ; Hodson, Harold F. ; Garland, Lawrence G.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-p3312-19158922ba98a2c0dd0912a018558c7e014dba94c673151d99456770769898313</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1991</creationdate><topic>Aniline Compounds</topic><topic>Biological and medical sciences</topic><topic>Calcium - metabolism</topic><topic>demethoxyvirdin</topic><topic>Fluorescence</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Fundamental immunology</topic><topic>Fura-2</topic><topic>Humans</topic><topic>Immunobiology</topic><topic>In Vitro Techniques</topic><topic>Inositol 1,4,5-Trisphosphate - metabolism</topic><topic>Myeloid cells: ontogeny, maturation, markers, receptors</topic><topic>N-Formylmethionine Leucyl-Phenylalanine - pharmacology</topic><topic>Neutrophils - drug effects</topic><topic>Neutrophils - metabolism</topic><topic>Phosphodiesterase Inhibitors - pharmacology</topic><topic>phospholipase C</topic><topic>phospholipase D</topic><topic>phospholipase inhibitor</topic><topic>Polynuclears</topic><topic>Receptors, Formyl Peptide</topic><topic>Receptors, Immunologic - drug effects</topic><topic>Receptors, Immunologic - metabolism</topic><topic>Signal transduction</topic><topic>Signal Transduction - drug effects</topic><topic>Xanthenes</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Thompson, Neil T.</creatorcontrib><creatorcontrib>Bonser, Robert W.</creatorcontrib><creatorcontrib>Tateson, James E.</creatorcontrib><creatorcontrib>Spacey, Graham D.</creatorcontrib><creatorcontrib>Randall, Roger W.</creatorcontrib><creatorcontrib>Hodson, Harold F.</creatorcontrib><creatorcontrib>Garland, Lawrence G.</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>MEDLINE - Academic</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>Thompson, Neil T.</au><au>Bonser, Robert W.</au><au>Tateson, James E.</au><au>Spacey, Graham D.</au><au>Randall, Roger W.</au><au>Hodson, Harold F.</au><au>Garland, Lawrence G.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A quantitative investigation into the dependence of Ca2+ mobilisation on changes in inositol 1,4,5‐trisphosphate levels in the stimulated neutrophil</atitle><jtitle>British journal of pharmacology</jtitle><addtitle>Br J Pharmacol</addtitle><date>1991-06</date><risdate>1991</risdate><volume>103</volume><issue>2</issue><spage>1592</spage><epage>1596</epage><pages>1592-1596</pages><issn>0007-1188</issn><eissn>1476-5381</eissn><coden>BJPCBM</coden><abstract>1 The coupling of N‐formyl‐methionyl‐leucyl‐phenylalanine (fMet‐Leu‐Phe) receptor stimulation to Ca2+ mobilisation has been investigated in the human neutrophil by measuring the concentration‐effect curves for inositol 1,4,5‐trisphosphate (IP3) formation and Ca2+ mobilisation. 2 fMet‐Leu‐Phe‐dependent mobilisation of intracellular Ca2+ has been monitored in fluo‐3‐loaded human neutrophils by measuring increases in the cytoplasmic free Ca2+ concentration ([Ca2+]i) in the presence of extracellular EGTA. Fluo‐3 was used in preference to fura‐2 because it was found to be more sensitive to the high Ca2+ levels seen in stimulated neutrophils. 3 fMet‐Leu‐Phe induced a rapid mobilisation of intracellular Ca2+ (EC50 = 2.9 ± 0.1 nm) and increased [Ca2+]i to a maximum of 1286 ± 184 nm. 4 The amount of IP3 in fMet‐Leu‐Phe‐stimulated neutrophils was determined by competition with [3H]‐IP3 for a specific IP3 binding protein isolated from bovine adrenocortical microsomes. Basal IP3 levels of 13.3 ± 2.0 pmol per 107 cells were increased nearly 4 fold by maximally effective concentrations of fMet‐Leu‐Phe. 5 The EC50 for the IP3 response (95 ± 18 nm) was much higher than that for mobilisation of intracellular Ca2+, such that only a doubling in the concentration of IP3 was required to fully mobilise intracellular Ca2+. 6 As a result of this relationship IP3 production was more sensitive than Ca2+ mobilisation to inhibition by demethoxyviridin, an inhibitor of phospholipase activation.</abstract><cop>Oxford, UK</cop><pub>Blackwell Publishing Ltd</pub><pmid>1884113</pmid><doi>10.1111/j.1476-5381.1991.tb09832.x</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record>
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source MEDLINE; EZB-FREE-00999 freely available EZB journals; PubMed Central; Alma/SFX Local Collection
subjects Aniline Compounds
Biological and medical sciences
Calcium - metabolism
demethoxyvirdin
Fluorescence
Fundamental and applied biological sciences. Psychology
Fundamental immunology
Fura-2
Humans
Immunobiology
In Vitro Techniques
Inositol 1,4,5-Trisphosphate - metabolism
Myeloid cells: ontogeny, maturation, markers, receptors
N-Formylmethionine Leucyl-Phenylalanine - pharmacology
Neutrophils - drug effects
Neutrophils - metabolism
Phosphodiesterase Inhibitors - pharmacology
phospholipase C
phospholipase D
phospholipase inhibitor
Polynuclears
Receptors, Formyl Peptide
Receptors, Immunologic - drug effects
Receptors, Immunologic - metabolism
Signal transduction
Signal Transduction - drug effects
Xanthenes
title A quantitative investigation into the dependence of Ca2+ mobilisation on changes in inositol 1,4,5‐trisphosphate levels in the stimulated neutrophil
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