A Fluorescence-Based Method for Measuring Nitric Oxide in Extracts of Skeletal Muscle
We describe here a fluorescence assay for nitric oxide synthase activity in skeletal muscle based on a new indicator, 4,5-diaminofluorescein (DAF-2). The rapid and irreversible binding of DAF-2 to oxidized NO allows real-time measurement of NO production. The method is safer and more convenient than...
Gespeichert in:
| Veröffentlicht in: | Nitric oxide 2001-10, Vol.5 (5), p.475-481 |
|---|---|
| Hauptverfasser: | , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 481 |
|---|---|
| container_issue | 5 |
| container_start_page | 475 |
| container_title | Nitric oxide |
| container_volume | 5 |
| creator | Sutherland, Hazel Khundkar, Roba Zolle, Olga McArdle, Anne Simpson, Alec W.M. Jarvis, Jonathan C. Salmons, Stanley |
| description | We describe here a fluorescence assay for nitric oxide synthase activity in skeletal muscle based on a new indicator, 4,5-diaminofluorescein (DAF-2). The rapid and irreversible binding of DAF-2 to oxidized NO allows real-time measurement of NO production. The method is safer and more convenient than the usual citrulline radioassay and can be used with crude muscle extracts. Rabbit fast tibialis anterior (TA) muscle had a nitric oxide synthase (NOS) activity of 44.3 ± 3.5 pmol/min/mg muscle. Addition of NOS blocker NG-allyl-l-arginine reduced this activity by 43%. Slow soleus muscle displayed NOS activity of 7.3 ± 2.5 pmol/min/mg muscle, 16% that of the TA muscle. Continuous stimulation of TA muscle at 10 Hz for 3 weeks reduced NOS activity by 47% to an intermediate value consistent with the associated conversion of the muscle phenotype from fast to slow. |
| doi_str_mv | 10.1006/niox.2001.0374 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_72178938</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S108986030190374X</els_id><sourcerecordid>72178938</sourcerecordid><originalsourceid>FETCH-LOGICAL-c406t-d4aacd5794c8acdb8a3df6a4c11af5e8042459f7057388d556cf19a6c388067a3</originalsourceid><addsrcrecordid>eNp1kDFPwzAQRi0EoqWwMiJPbAl24zjOWKoWkAodoLPl2hcwpHGxE1T-PY5awcR030nvPukeQpeUpJQQftNYt0vHhNCUZAU7QkNKRJkITunxbybZAJ2F8E4IYZngp2hAaS6KnI-HaDXB87pzHoKGRkNyqwIY_AjtmzO4cj5GFTpvm1f8ZFtvNV7urAFsGzzbtV7pNmBX4ecPqKFVNX7sgq7hHJ1Uqg5wcZgjtJrPXqb3yWJ59zCdLBLNCG8Tw5TSJi9KpkUMa6EyU3HFNKWqykEQNmZ5WRUkLzIhTJ5zXdFScR03wguVjdD1vnfr3WcHoZUbGx-pa9WA64IsxrQQZSYimO5B7V0IHiq59Xaj_LekRPYiZS9S9iJlLzIeXB2au_UGzB9-MBcBsQcg_vdlwcugba_QWA-6lcbZ_7p_APMJgfg</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>72178938</pqid></control><display><type>article</type><title>A Fluorescence-Based Method for Measuring Nitric Oxide in Extracts of Skeletal Muscle</title><source>MEDLINE</source><source>Access via ScienceDirect (Elsevier)</source><creator>Sutherland, Hazel ; Khundkar, Roba ; Zolle, Olga ; McArdle, Anne ; Simpson, Alec W.M. ; Jarvis, Jonathan C. ; Salmons, Stanley</creator><creatorcontrib>Sutherland, Hazel ; Khundkar, Roba ; Zolle, Olga ; McArdle, Anne ; Simpson, Alec W.M. ; Jarvis, Jonathan C. ; Salmons, Stanley</creatorcontrib><description>We describe here a fluorescence assay for nitric oxide synthase activity in skeletal muscle based on a new indicator, 4,5-diaminofluorescein (DAF-2). The rapid and irreversible binding of DAF-2 to oxidized NO allows real-time measurement of NO production. The method is safer and more convenient than the usual citrulline radioassay and can be used with crude muscle extracts. Rabbit fast tibialis anterior (TA) muscle had a nitric oxide synthase (NOS) activity of 44.3 ± 3.5 pmol/min/mg muscle. Addition of NOS blocker NG-allyl-l-arginine reduced this activity by 43%. Slow soleus muscle displayed NOS activity of 7.3 ± 2.5 pmol/min/mg muscle, 16% that of the TA muscle. Continuous stimulation of TA muscle at 10 Hz for 3 weeks reduced NOS activity by 47% to an intermediate value consistent with the associated conversion of the muscle phenotype from fast to slow.</description><identifier>ISSN: 1089-8603</identifier><identifier>EISSN: 1089-8611</identifier><identifier>DOI: 10.1006/niox.2001.0374</identifier><identifier>PMID: 11587562</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>[formula omitted]-NMMA ; Animals ; DAF-2 ; Fluorescein ; Indicators and Reagents ; Mice ; Mice, Inbred C57BL ; Mice, Inbred mdx ; Muscle, Skeletal - chemistry ; NG-allyl-[formula omitted]-arginine ; nitric oxide (NO) ; Nitric Oxide - analysis ; nitric oxide synthase (NOS) ; Nitric Oxide Synthase - antagonists & inhibitors ; Rabbits ; Sensitivity and Specificity ; skeletal muscle ; Spectrometry, Fluorescence - methods ; transformation</subject><ispartof>Nitric oxide, 2001-10, Vol.5 (5), p.475-481</ispartof><rights>2001 Academic Press</rights><rights>Copyright 2001 Academic Press.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c406t-d4aacd5794c8acdb8a3df6a4c11af5e8042459f7057388d556cf19a6c388067a3</citedby><cites>FETCH-LOGICAL-c406t-d4aacd5794c8acdb8a3df6a4c11af5e8042459f7057388d556cf19a6c388067a3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1006/niox.2001.0374$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>315,781,785,3551,27929,27930,46000</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11587562$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Sutherland, Hazel</creatorcontrib><creatorcontrib>Khundkar, Roba</creatorcontrib><creatorcontrib>Zolle, Olga</creatorcontrib><creatorcontrib>McArdle, Anne</creatorcontrib><creatorcontrib>Simpson, Alec W.M.</creatorcontrib><creatorcontrib>Jarvis, Jonathan C.</creatorcontrib><creatorcontrib>Salmons, Stanley</creatorcontrib><title>A Fluorescence-Based Method for Measuring Nitric Oxide in Extracts of Skeletal Muscle</title><title>Nitric oxide</title><addtitle>Nitric Oxide</addtitle><description>We describe here a fluorescence assay for nitric oxide synthase activity in skeletal muscle based on a new indicator, 4,5-diaminofluorescein (DAF-2). The rapid and irreversible binding of DAF-2 to oxidized NO allows real-time measurement of NO production. The method is safer and more convenient than the usual citrulline radioassay and can be used with crude muscle extracts. Rabbit fast tibialis anterior (TA) muscle had a nitric oxide synthase (NOS) activity of 44.3 ± 3.5 pmol/min/mg muscle. Addition of NOS blocker NG-allyl-l-arginine reduced this activity by 43%. Slow soleus muscle displayed NOS activity of 7.3 ± 2.5 pmol/min/mg muscle, 16% that of the TA muscle. Continuous stimulation of TA muscle at 10 Hz for 3 weeks reduced NOS activity by 47% to an intermediate value consistent with the associated conversion of the muscle phenotype from fast to slow.</description><subject>[formula omitted]-NMMA</subject><subject>Animals</subject><subject>DAF-2</subject><subject>Fluorescein</subject><subject>Indicators and Reagents</subject><subject>Mice</subject><subject>Mice, Inbred C57BL</subject><subject>Mice, Inbred mdx</subject><subject>Muscle, Skeletal - chemistry</subject><subject>NG-allyl-[formula omitted]-arginine</subject><subject>nitric oxide (NO)</subject><subject>Nitric Oxide - analysis</subject><subject>nitric oxide synthase (NOS)</subject><subject>Nitric Oxide Synthase - antagonists & inhibitors</subject><subject>Rabbits</subject><subject>Sensitivity and Specificity</subject><subject>skeletal muscle</subject><subject>Spectrometry, Fluorescence - methods</subject><subject>transformation</subject><issn>1089-8603</issn><issn>1089-8611</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2001</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kDFPwzAQRi0EoqWwMiJPbAl24zjOWKoWkAodoLPl2hcwpHGxE1T-PY5awcR030nvPukeQpeUpJQQftNYt0vHhNCUZAU7QkNKRJkITunxbybZAJ2F8E4IYZngp2hAaS6KnI-HaDXB87pzHoKGRkNyqwIY_AjtmzO4cj5GFTpvm1f8ZFtvNV7urAFsGzzbtV7pNmBX4ecPqKFVNX7sgq7hHJ1Uqg5wcZgjtJrPXqb3yWJ59zCdLBLNCG8Tw5TSJi9KpkUMa6EyU3HFNKWqykEQNmZ5WRUkLzIhTJ5zXdFScR03wguVjdD1vnfr3WcHoZUbGx-pa9WA64IsxrQQZSYimO5B7V0IHiq59Xaj_LekRPYiZS9S9iJlLzIeXB2au_UGzB9-MBcBsQcg_vdlwcugba_QWA-6lcbZ_7p_APMJgfg</recordid><startdate>20011001</startdate><enddate>20011001</enddate><creator>Sutherland, Hazel</creator><creator>Khundkar, Roba</creator><creator>Zolle, Olga</creator><creator>McArdle, Anne</creator><creator>Simpson, Alec W.M.</creator><creator>Jarvis, Jonathan C.</creator><creator>Salmons, Stanley</creator><general>Elsevier Inc</general><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></search><sort><creationdate>20011001</creationdate><title>A Fluorescence-Based Method for Measuring Nitric Oxide in Extracts of Skeletal Muscle</title><author>Sutherland, Hazel ; Khundkar, Roba ; Zolle, Olga ; McArdle, Anne ; Simpson, Alec W.M. ; Jarvis, Jonathan C. ; Salmons, Stanley</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c406t-d4aacd5794c8acdb8a3df6a4c11af5e8042459f7057388d556cf19a6c388067a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2001</creationdate><topic>[formula omitted]-NMMA</topic><topic>Animals</topic><topic>DAF-2</topic><topic>Fluorescein</topic><topic>Indicators and Reagents</topic><topic>Mice</topic><topic>Mice, Inbred C57BL</topic><topic>Mice, Inbred mdx</topic><topic>Muscle, Skeletal - chemistry</topic><topic>NG-allyl-[formula omitted]-arginine</topic><topic>nitric oxide (NO)</topic><topic>Nitric Oxide - analysis</topic><topic>nitric oxide synthase (NOS)</topic><topic>Nitric Oxide Synthase - antagonists & inhibitors</topic><topic>Rabbits</topic><topic>Sensitivity and Specificity</topic><topic>skeletal muscle</topic><topic>Spectrometry, Fluorescence - methods</topic><topic>transformation</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Sutherland, Hazel</creatorcontrib><creatorcontrib>Khundkar, Roba</creatorcontrib><creatorcontrib>Zolle, Olga</creatorcontrib><creatorcontrib>McArdle, Anne</creatorcontrib><creatorcontrib>Simpson, Alec W.M.</creatorcontrib><creatorcontrib>Jarvis, Jonathan C.</creatorcontrib><creatorcontrib>Salmons, Stanley</creatorcontrib><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><jtitle>Nitric oxide</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Sutherland, Hazel</au><au>Khundkar, Roba</au><au>Zolle, Olga</au><au>McArdle, Anne</au><au>Simpson, Alec W.M.</au><au>Jarvis, Jonathan C.</au><au>Salmons, Stanley</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A Fluorescence-Based Method for Measuring Nitric Oxide in Extracts of Skeletal Muscle</atitle><jtitle>Nitric oxide</jtitle><addtitle>Nitric Oxide</addtitle><date>2001-10-01</date><risdate>2001</risdate><volume>5</volume><issue>5</issue><spage>475</spage><epage>481</epage><pages>475-481</pages><issn>1089-8603</issn><eissn>1089-8611</eissn><abstract>We describe here a fluorescence assay for nitric oxide synthase activity in skeletal muscle based on a new indicator, 4,5-diaminofluorescein (DAF-2). The rapid and irreversible binding of DAF-2 to oxidized NO allows real-time measurement of NO production. The method is safer and more convenient than the usual citrulline radioassay and can be used with crude muscle extracts. Rabbit fast tibialis anterior (TA) muscle had a nitric oxide synthase (NOS) activity of 44.3 ± 3.5 pmol/min/mg muscle. Addition of NOS blocker NG-allyl-l-arginine reduced this activity by 43%. Slow soleus muscle displayed NOS activity of 7.3 ± 2.5 pmol/min/mg muscle, 16% that of the TA muscle. Continuous stimulation of TA muscle at 10 Hz for 3 weeks reduced NOS activity by 47% to an intermediate value consistent with the associated conversion of the muscle phenotype from fast to slow.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>11587562</pmid><doi>10.1006/niox.2001.0374</doi><tpages>7</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1089-8603 |
| ispartof | Nitric oxide, 2001-10, Vol.5 (5), p.475-481 |
| issn | 1089-8603 1089-8611 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_72178938 |
| source | MEDLINE; Access via ScienceDirect (Elsevier) |
| subjects | [formula omitted]-NMMA Animals DAF-2 Fluorescein Indicators and Reagents Mice Mice, Inbred C57BL Mice, Inbred mdx Muscle, Skeletal - chemistry NG-allyl-[formula omitted]-arginine nitric oxide (NO) Nitric Oxide - analysis nitric oxide synthase (NOS) Nitric Oxide Synthase - antagonists & inhibitors Rabbits Sensitivity and Specificity skeletal muscle Spectrometry, Fluorescence - methods transformation |
| title | A Fluorescence-Based Method for Measuring Nitric Oxide in Extracts of Skeletal Muscle |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-12T11%3A41%3A15IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=A%20Fluorescence-Based%20Method%20for%20Measuring%20Nitric%20Oxide%20in%20Extracts%20of%20Skeletal%20Muscle&rft.jtitle=Nitric%20oxide&rft.au=Sutherland,%20Hazel&rft.date=2001-10-01&rft.volume=5&rft.issue=5&rft.spage=475&rft.epage=481&rft.pages=475-481&rft.issn=1089-8603&rft.eissn=1089-8611&rft_id=info:doi/10.1006/niox.2001.0374&rft_dat=%3Cproquest_cross%3E72178938%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=72178938&rft_id=info:pmid/11587562&rft_els_id=S108986030190374X&rfr_iscdi=true |