Measurement of the internal pH of mast cell granules using microvolumetric fluorescence and isotopic techniques
The intragranular pH of isolated mast cell granules was measured. Because of the minute amounts of isolated granules available, two techniques were developed by modifying aminoacridine fluorescence and [ 14C]methylamine accumulation techniques to permit measurements with microliter sample volumes. G...
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Veröffentlicht in: | Arch. Biochem. Biophys.; (United States) 1987-04, Vol.254 (1), p.222-233 |
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description | The intragranular pH of isolated mast cell granules was measured. Because of the minute amounts of isolated granules available, two techniques were developed by modifying aminoacridine fluorescence and [
14C]methylamine accumulation techniques to permit measurements with microliter sample volumes. Granule purity was demonstrated by electron microscopy, ruthenium red exclusion, and biochemical (histamine, mast cell granule protease) analysis. The internal pH was determined to be 5.55 ± 0.06, indicating that the pH environment within mast cell granules is not significantly different from that of previously studied granule types (i.e., chromaffin, platelet, pancreatic islet, and pituitary granules). Collapse of the pH gradient by NH
4
+ was demonstrated with both techniques. No evidence of
Cl
−
OH
−
or specific cation/H
+ transport was found, and major chloride permeability could not be unequivocably demonstrated. Ca
2+ and Cl
− at concentrations normally present extracellularly destabilized granules in the presence of NH
4
+, but this phenomenon does not necessarily indicate a role for these ions in the exocytotic release of granule contents from intact cells. The pH measurement techniques developed for investigating the properties of granules in mast cells may be useful for studying other granules that can be obtained only in limited quantities. |
doi_str_mv | 10.1016/0003-9861(87)90098-1 |
format | Article |
fullrecord | <record><control><sourceid>proquest_osti_</sourceid><recordid>TN_cdi_proquest_miscellaneous_77513968</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>0003986187900981</els_id><sourcerecordid>77513968</sourcerecordid><originalsourceid>FETCH-LOGICAL-c479t-da4607f5032031f364315fad65ac9f0e77b0e0f5fc32aed4ac06a4c2696db653</originalsourceid><addsrcrecordid>eNp9kUFrFTEUhYMo9dn6DxSCiOhi7M1kJplsClJaK7S46T7kZW7ayEzyTDIF_30nfY-3dBXI-e7lnHMJ-cDgOwMmzgGAN2oQ7OsgvykANTTsFdkwUKIBPnSvyeaIvCXvcv4DwFgn2hNywnupWqU2JN6hyUvCGUOh0dHyiNSHgimYie5u6tdscqEWp4k-JBOWCTNdsg8PdPY2xac4LTOW5C110xITZovBIjVhpD7HEnerUtA-Bv93wXxG3jgzZXx_eE_J_fXV_eVNc_v756_LH7eN7aQqzWg6AdL1wFvgzHHRcdY7M4reWOUApdwCguud5a3BsTMWhOlsK5QYt6Lnp-TTfm3MxetsfXVgYwhoixYdqFYMK_RlD-1SrN6Knn2uQU3AuGQtZc-4egG7PbjmzTmh07vkZ5P-aQa63kLXonUtWg9Sv9xCs3Xs42H_sp1xPA4dyl_1zwfdZGsmt7ZrfT5ishfQ9nzFLvYYrn09eUw1T-149KnGGaP_v49n5UKm2Q</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>77513968</pqid></control><display><type>article</type><title>Measurement of the internal pH of mast cell granules using microvolumetric fluorescence and isotopic techniques</title><source>MEDLINE</source><source>Elsevier ScienceDirect Journals</source><creator>De Young, Mary Beth ; Nemeth, Edward F. ; Scarpa, Antonio</creator><creatorcontrib>De Young, Mary Beth ; Nemeth, Edward F. ; Scarpa, Antonio ; Case Western Reserve Univ. School of Medicine, Cleveland, OH</creatorcontrib><description>The intragranular pH of isolated mast cell granules was measured. Because of the minute amounts of isolated granules available, two techniques were developed by modifying aminoacridine fluorescence and [
14C]methylamine accumulation techniques to permit measurements with microliter sample volumes. Granule purity was demonstrated by electron microscopy, ruthenium red exclusion, and biochemical (histamine, mast cell granule protease) analysis. The internal pH was determined to be 5.55 ± 0.06, indicating that the pH environment within mast cell granules is not significantly different from that of previously studied granule types (i.e., chromaffin, platelet, pancreatic islet, and pituitary granules). Collapse of the pH gradient by NH
4
+ was demonstrated with both techniques. No evidence of
Cl
−
OH
−
or specific cation/H
+ transport was found, and major chloride permeability could not be unequivocably demonstrated. Ca
2+ and Cl
− at concentrations normally present extracellularly destabilized granules in the presence of NH
4
+, but this phenomenon does not necessarily indicate a role for these ions in the exocytotic release of granule contents from intact cells. The pH measurement techniques developed for investigating the properties of granules in mast cells may be useful for studying other granules that can be obtained only in limited quantities.</description><identifier>ISSN: 0003-9861</identifier><identifier>EISSN: 1096-0384</identifier><identifier>DOI: 10.1016/0003-9861(87)90098-1</identifier><identifier>PMID: 3579299</identifier><identifier>CODEN: ABBIA4</identifier><language>eng</language><publisher>San Diego, CA: Elsevier Inc</publisher><subject>550201 - Biochemistry- Tracer Techniques ; AMINES ; Aminoacridines ; ANIMAL CELLS ; ANIMALS ; Applied sciences ; BASIC BIOLOGICAL SCIENCES ; CARBON 14 COMPOUNDS ; Carbon Radioisotopes ; CONNECTIVE TISSUE CELLS ; Cytoplasmic Granules - metabolism ; Exact sciences and technology ; FLUORESCENCE ; Hydrogen-Ion Concentration ; Ions ; ISOTOPE APPLICATIONS ; LABELLED COMPOUNDS ; LUMINESCENCE ; Male ; MAMMALS ; MAST CELLS ; Mast Cells - metabolism ; MEASURING METHODS ; METHYLAMINE ; Methylamines - metabolism ; ORGANIC COMPOUNDS ; Other techniques and industries ; PH VALUE ; RATS ; RESPONSE MODIFYING FACTORS ; RODENTS ; SOMATIC CELLS ; Spectrometry, Fluorescence ; TRACER TECHNIQUES ; VERTEBRATES</subject><ispartof>Arch. Biochem. Biophys.; (United States), 1987-04, Vol.254 (1), p.222-233</ispartof><rights>1987</rights><rights>1988 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c479t-da4607f5032031f364315fad65ac9f0e77b0e0f5fc32aed4ac06a4c2696db653</citedby><cites>FETCH-LOGICAL-c479t-da4607f5032031f364315fad65ac9f0e77b0e0f5fc32aed4ac06a4c2696db653</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/0003986187900981$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,881,3537,27903,27904,65309</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=7560253$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/3579299$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://www.osti.gov/biblio/6409268$$D View this record in Osti.gov$$Hfree_for_read</backlink></links><search><creatorcontrib>De Young, Mary Beth</creatorcontrib><creatorcontrib>Nemeth, Edward F.</creatorcontrib><creatorcontrib>Scarpa, Antonio</creatorcontrib><creatorcontrib>Case Western Reserve Univ. School of Medicine, Cleveland, OH</creatorcontrib><title>Measurement of the internal pH of mast cell granules using microvolumetric fluorescence and isotopic techniques</title><title>Arch. Biochem. Biophys.; (United States)</title><addtitle>Arch Biochem Biophys</addtitle><description>The intragranular pH of isolated mast cell granules was measured. Because of the minute amounts of isolated granules available, two techniques were developed by modifying aminoacridine fluorescence and [
14C]methylamine accumulation techniques to permit measurements with microliter sample volumes. Granule purity was demonstrated by electron microscopy, ruthenium red exclusion, and biochemical (histamine, mast cell granule protease) analysis. The internal pH was determined to be 5.55 ± 0.06, indicating that the pH environment within mast cell granules is not significantly different from that of previously studied granule types (i.e., chromaffin, platelet, pancreatic islet, and pituitary granules). Collapse of the pH gradient by NH
4
+ was demonstrated with both techniques. No evidence of
Cl
−
OH
−
or specific cation/H
+ transport was found, and major chloride permeability could not be unequivocably demonstrated. Ca
2+ and Cl
− at concentrations normally present extracellularly destabilized granules in the presence of NH
4
+, but this phenomenon does not necessarily indicate a role for these ions in the exocytotic release of granule contents from intact cells. The pH measurement techniques developed for investigating the properties of granules in mast cells may be useful for studying other granules that can be obtained only in limited quantities.</description><subject>550201 - Biochemistry- Tracer Techniques</subject><subject>AMINES</subject><subject>Aminoacridines</subject><subject>ANIMAL CELLS</subject><subject>ANIMALS</subject><subject>Applied sciences</subject><subject>BASIC BIOLOGICAL SCIENCES</subject><subject>CARBON 14 COMPOUNDS</subject><subject>Carbon Radioisotopes</subject><subject>CONNECTIVE TISSUE CELLS</subject><subject>Cytoplasmic Granules - metabolism</subject><subject>Exact sciences and technology</subject><subject>FLUORESCENCE</subject><subject>Hydrogen-Ion Concentration</subject><subject>Ions</subject><subject>ISOTOPE APPLICATIONS</subject><subject>LABELLED COMPOUNDS</subject><subject>LUMINESCENCE</subject><subject>Male</subject><subject>MAMMALS</subject><subject>MAST CELLS</subject><subject>Mast Cells - metabolism</subject><subject>MEASURING METHODS</subject><subject>METHYLAMINE</subject><subject>Methylamines - metabolism</subject><subject>ORGANIC COMPOUNDS</subject><subject>Other techniques and industries</subject><subject>PH VALUE</subject><subject>RATS</subject><subject>RESPONSE MODIFYING FACTORS</subject><subject>RODENTS</subject><subject>SOMATIC CELLS</subject><subject>Spectrometry, Fluorescence</subject><subject>TRACER TECHNIQUES</subject><subject>VERTEBRATES</subject><issn>0003-9861</issn><issn>1096-0384</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1987</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kUFrFTEUhYMo9dn6DxSCiOhi7M1kJplsClJaK7S46T7kZW7ayEzyTDIF_30nfY-3dBXI-e7lnHMJ-cDgOwMmzgGAN2oQ7OsgvykANTTsFdkwUKIBPnSvyeaIvCXvcv4DwFgn2hNywnupWqU2JN6hyUvCGUOh0dHyiNSHgimYie5u6tdscqEWp4k-JBOWCTNdsg8PdPY2xac4LTOW5C110xITZovBIjVhpD7HEnerUtA-Bv93wXxG3jgzZXx_eE_J_fXV_eVNc_v756_LH7eN7aQqzWg6AdL1wFvgzHHRcdY7M4reWOUApdwCguud5a3BsTMWhOlsK5QYt6Lnp-TTfm3MxetsfXVgYwhoixYdqFYMK_RlD-1SrN6Knn2uQU3AuGQtZc-4egG7PbjmzTmh07vkZ5P-aQa63kLXonUtWg9Sv9xCs3Xs42H_sp1xPA4dyl_1zwfdZGsmt7ZrfT5ishfQ9nzFLvYYrn09eUw1T-149KnGGaP_v49n5UKm2Q</recordid><startdate>19870401</startdate><enddate>19870401</enddate><creator>De Young, Mary Beth</creator><creator>Nemeth, Edward F.</creator><creator>Scarpa, Antonio</creator><general>Elsevier Inc</general><general>Elsevier</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>7X8</scope><scope>OTOTI</scope></search><sort><creationdate>19870401</creationdate><title>Measurement of the internal pH of mast cell granules using microvolumetric fluorescence and isotopic techniques</title><author>De Young, Mary Beth ; Nemeth, Edward F. ; Scarpa, Antonio</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c479t-da4607f5032031f364315fad65ac9f0e77b0e0f5fc32aed4ac06a4c2696db653</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1987</creationdate><topic>550201 - Biochemistry- Tracer Techniques</topic><topic>AMINES</topic><topic>Aminoacridines</topic><topic>ANIMAL CELLS</topic><topic>ANIMALS</topic><topic>Applied sciences</topic><topic>BASIC BIOLOGICAL SCIENCES</topic><topic>CARBON 14 COMPOUNDS</topic><topic>Carbon Radioisotopes</topic><topic>CONNECTIVE TISSUE CELLS</topic><topic>Cytoplasmic Granules - metabolism</topic><topic>Exact sciences and technology</topic><topic>FLUORESCENCE</topic><topic>Hydrogen-Ion Concentration</topic><topic>Ions</topic><topic>ISOTOPE APPLICATIONS</topic><topic>LABELLED COMPOUNDS</topic><topic>LUMINESCENCE</topic><topic>Male</topic><topic>MAMMALS</topic><topic>MAST CELLS</topic><topic>Mast Cells - metabolism</topic><topic>MEASURING METHODS</topic><topic>METHYLAMINE</topic><topic>Methylamines - metabolism</topic><topic>ORGANIC COMPOUNDS</topic><topic>Other techniques and industries</topic><topic>PH VALUE</topic><topic>RATS</topic><topic>RESPONSE MODIFYING FACTORS</topic><topic>RODENTS</topic><topic>SOMATIC CELLS</topic><topic>Spectrometry, Fluorescence</topic><topic>TRACER TECHNIQUES</topic><topic>VERTEBRATES</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>De Young, Mary Beth</creatorcontrib><creatorcontrib>Nemeth, Edward F.</creatorcontrib><creatorcontrib>Scarpa, Antonio</creatorcontrib><creatorcontrib>Case Western Reserve Univ. School of Medicine, Cleveland, OH</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>MEDLINE - Academic</collection><collection>OSTI.GOV</collection><jtitle>Arch. Biochem. Biophys.; (United States)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>De Young, Mary Beth</au><au>Nemeth, Edward F.</au><au>Scarpa, Antonio</au><aucorp>Case Western Reserve Univ. School of Medicine, Cleveland, OH</aucorp><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Measurement of the internal pH of mast cell granules using microvolumetric fluorescence and isotopic techniques</atitle><jtitle>Arch. Biochem. Biophys.; (United States)</jtitle><addtitle>Arch Biochem Biophys</addtitle><date>1987-04-01</date><risdate>1987</risdate><volume>254</volume><issue>1</issue><spage>222</spage><epage>233</epage><pages>222-233</pages><issn>0003-9861</issn><eissn>1096-0384</eissn><coden>ABBIA4</coden><abstract>The intragranular pH of isolated mast cell granules was measured. Because of the minute amounts of isolated granules available, two techniques were developed by modifying aminoacridine fluorescence and [
14C]methylamine accumulation techniques to permit measurements with microliter sample volumes. Granule purity was demonstrated by electron microscopy, ruthenium red exclusion, and biochemical (histamine, mast cell granule protease) analysis. The internal pH was determined to be 5.55 ± 0.06, indicating that the pH environment within mast cell granules is not significantly different from that of previously studied granule types (i.e., chromaffin, platelet, pancreatic islet, and pituitary granules). Collapse of the pH gradient by NH
4
+ was demonstrated with both techniques. No evidence of
Cl
−
OH
−
or specific cation/H
+ transport was found, and major chloride permeability could not be unequivocably demonstrated. Ca
2+ and Cl
− at concentrations normally present extracellularly destabilized granules in the presence of NH
4
+, but this phenomenon does not necessarily indicate a role for these ions in the exocytotic release of granule contents from intact cells. The pH measurement techniques developed for investigating the properties of granules in mast cells may be useful for studying other granules that can be obtained only in limited quantities.</abstract><cop>San Diego, CA</cop><pub>Elsevier Inc</pub><pmid>3579299</pmid><doi>10.1016/0003-9861(87)90098-1</doi><tpages>12</tpages></addata></record> |
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source | MEDLINE; Elsevier ScienceDirect Journals |
subjects | 550201 - Biochemistry- Tracer Techniques AMINES Aminoacridines ANIMAL CELLS ANIMALS Applied sciences BASIC BIOLOGICAL SCIENCES CARBON 14 COMPOUNDS Carbon Radioisotopes CONNECTIVE TISSUE CELLS Cytoplasmic Granules - metabolism Exact sciences and technology FLUORESCENCE Hydrogen-Ion Concentration Ions ISOTOPE APPLICATIONS LABELLED COMPOUNDS LUMINESCENCE Male MAMMALS MAST CELLS Mast Cells - metabolism MEASURING METHODS METHYLAMINE Methylamines - metabolism ORGANIC COMPOUNDS Other techniques and industries PH VALUE RATS RESPONSE MODIFYING FACTORS RODENTS SOMATIC CELLS Spectrometry, Fluorescence TRACER TECHNIQUES VERTEBRATES |
title | Measurement of the internal pH of mast cell granules using microvolumetric fluorescence and isotopic techniques |
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