The use of CalciumOrange-5N as a specific marker of mitochondrial Ca2+ in mouse skeletal muscle fibers
We report the use of the fluorescent dye CalciumOrange-5N (CaOr-5N) as a specific mitochondria Ca 2+ marker in enzymatically dissociated mouse FBD muscle fibers. Using laser scanning confocal microscopy and the dyes Mitotracker Green (MTG), di-8-ANEPPS and endoplasmic reticulum tracker green (ERTG),...
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| Veröffentlicht in: | Pflügers Archiv 2008-01, Vol.455 (4), p.721-731 |
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| creator | Bolaños, Pura Guillen, Alis Rojas, Héctor Boncompagni, Simona Caputo, Carlo |
| description | We report the use of the fluorescent dye CalciumOrange-5N (CaOr-5N) as a specific mitochondria Ca
2+
marker in enzymatically dissociated mouse FBD muscle fibers. Using laser scanning confocal microscopy and the dyes Mitotracker Green (MTG), di-8-ANEPPS and endoplasmic reticulum tracker green (ERTG), we determined the relative position of mitochondria, transverse tubules and sarcoplasmic reticulum in the sarcomere. Comparison with electron micrographies showed that mitochondria are mostly present at both sides of Z lines and near the triads located at the A-I band border. CaOr-5N fluorescence was mainly distributed in mitochondria, highly co-localised with MTG and basically excluded from the A band space. ERTG localised mostly between the two t-tubules present in each sarcomere. We studied the effect of the protonophore FCCP using CaOr-5N to measure mitochondrial Ca
2+
and JC-1 dye to measure mitochondria inner membrane potential (Δ
Ψ
m
). After FCCP treatment, the CaOr-5N fluorescence diminished by about 33% in 80 s, while JC-1 fluorescence diminished by 36% in 200 s. Our results show the loss of Ca
2+
from mitochondria when ΔΨ
m
is depolarised and demonstrate the usefulness of CaOr-5N to mark mitochondrial [Ca
2+
]
m
. |
| doi_str_mv | 10.1007/s00424-007-0312-5 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_70102257</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>1998439261</sourcerecordid><originalsourceid>FETCH-LOGICAL-c365t-4b79a75b0c8af4dee41cd3135ee1ec107eeb2aca7eda88cbf3dc479a5cda77ad3</originalsourceid><addsrcrecordid>eNp1kU1LAzEQhoMotn78AC8SPHiR6CSbbLZHKX6B2Es9h2x21kb3oybdg__eLC0UBE8ZJs_7zvAOIRccbjmAvosAUkiWSgYZF0wdkCmXmWACeHZIppC6LNd5MSEnMX4CgJCFOCYTrjUokPmU1MsV0iEi7Ws6t43zQ7sItvtApt6ojdTSuEbna-9oa8MXhhFs_aZ3q76rgrdNkokb6jva9qNP_MIGN6ndDtE1SGtfYohn5Ki2TcTz3XtK3h8flvNn9rp4epnfvzKX5WrDZKlnVqsSXGFrWSFK7qqMZwqRo-OgEUthndVY2aJwZZ1VTiaJcpXV2lbZKbne-q5D_z1g3JjWR4dNYztM6xkNHIRQOoFXf8DPfghd2s2IFBvMcg4J4lvIhT7GgLVZB59i-DEczHgBs72AGcvxAkYlzeXOeChbrPaKXeQJEFsgpq8UdNhP_t_1F6_jkTc</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>200309610</pqid></control><display><type>article</type><title>The use of CalciumOrange-5N as a specific marker of mitochondrial Ca2+ in mouse skeletal muscle fibers</title><source>MEDLINE</source><source>Springer Nature - Complete Springer Journals</source><creator>Bolaños, Pura ; Guillen, Alis ; Rojas, Héctor ; Boncompagni, Simona ; Caputo, Carlo</creator><creatorcontrib>Bolaños, Pura ; Guillen, Alis ; Rojas, Héctor ; Boncompagni, Simona ; Caputo, Carlo</creatorcontrib><description>We report the use of the fluorescent dye CalciumOrange-5N (CaOr-5N) as a specific mitochondria Ca
2+
marker in enzymatically dissociated mouse FBD muscle fibers. Using laser scanning confocal microscopy and the dyes Mitotracker Green (MTG), di-8-ANEPPS and endoplasmic reticulum tracker green (ERTG), we determined the relative position of mitochondria, transverse tubules and sarcoplasmic reticulum in the sarcomere. Comparison with electron micrographies showed that mitochondria are mostly present at both sides of Z lines and near the triads located at the A-I band border. CaOr-5N fluorescence was mainly distributed in mitochondria, highly co-localised with MTG and basically excluded from the A band space. ERTG localised mostly between the two t-tubules present in each sarcomere. We studied the effect of the protonophore FCCP using CaOr-5N to measure mitochondrial Ca
2+
and JC-1 dye to measure mitochondria inner membrane potential (Δ
Ψ
m
). After FCCP treatment, the CaOr-5N fluorescence diminished by about 33% in 80 s, while JC-1 fluorescence diminished by 36% in 200 s. Our results show the loss of Ca
2+
from mitochondria when ΔΨ
m
is depolarised and demonstrate the usefulness of CaOr-5N to mark mitochondrial [Ca
2+
]
m
.</description><identifier>ISSN: 0031-6768</identifier><identifier>EISSN: 1432-2013</identifier><identifier>DOI: 10.1007/s00424-007-0312-5</identifier><identifier>PMID: 17705046</identifier><language>eng</language><publisher>Berlin/Heidelberg: Springer-Verlag</publisher><subject>Aldehydes ; Animals ; Benzimidazoles ; Biomedical and Life Sciences ; Biomedicine ; Calcium ; Calcium - metabolism ; Carbocyanines ; Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone - pharmacology ; Cell Biology ; Dyes ; Fluorescent Dyes ; Human Physiology ; Membrane Potential, Mitochondrial - drug effects ; Mice ; Microscopy, Confocal ; Mitochondria ; Mitochondria, Muscle - metabolism ; Mitochondria, Muscle - ultrastructure ; Molecular Medicine ; Muscle Fibers, Skeletal - metabolism ; Muscle Fibers, Skeletal - ultrastructure ; Muscle, Skeletal - metabolism ; Muscle, Skeletal - ultrastructure ; Muscular system ; Neurosciences ; Organic Chemicals ; Proteins ; Pyridinium Compounds ; Receptors ; Sarcomeres - metabolism ; Sarcoplasmic Reticulum - metabolism ; Skeletal Muscle ; Staining and Labeling - methods ; Time Factors ; Uncoupling Agents - pharmacology</subject><ispartof>Pflügers Archiv, 2008-01, Vol.455 (4), p.721-731</ispartof><rights>Springer-Verlag 2007</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c365t-4b79a75b0c8af4dee41cd3135ee1ec107eeb2aca7eda88cbf3dc479a5cda77ad3</citedby><cites>FETCH-LOGICAL-c365t-4b79a75b0c8af4dee41cd3135ee1ec107eeb2aca7eda88cbf3dc479a5cda77ad3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s00424-007-0312-5$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s00424-007-0312-5$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,778,782,27907,27908,41471,42540,51302</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/17705046$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Bolaños, Pura</creatorcontrib><creatorcontrib>Guillen, Alis</creatorcontrib><creatorcontrib>Rojas, Héctor</creatorcontrib><creatorcontrib>Boncompagni, Simona</creatorcontrib><creatorcontrib>Caputo, Carlo</creatorcontrib><title>The use of CalciumOrange-5N as a specific marker of mitochondrial Ca2+ in mouse skeletal muscle fibers</title><title>Pflügers Archiv</title><addtitle>Pflugers Arch - Eur J Physiol</addtitle><addtitle>Pflugers Arch</addtitle><description>We report the use of the fluorescent dye CalciumOrange-5N (CaOr-5N) as a specific mitochondria Ca
2+
marker in enzymatically dissociated mouse FBD muscle fibers. Using laser scanning confocal microscopy and the dyes Mitotracker Green (MTG), di-8-ANEPPS and endoplasmic reticulum tracker green (ERTG), we determined the relative position of mitochondria, transverse tubules and sarcoplasmic reticulum in the sarcomere. Comparison with electron micrographies showed that mitochondria are mostly present at both sides of Z lines and near the triads located at the A-I band border. CaOr-5N fluorescence was mainly distributed in mitochondria, highly co-localised with MTG and basically excluded from the A band space. ERTG localised mostly between the two t-tubules present in each sarcomere. We studied the effect of the protonophore FCCP using CaOr-5N to measure mitochondrial Ca
2+
and JC-1 dye to measure mitochondria inner membrane potential (Δ
Ψ
m
). After FCCP treatment, the CaOr-5N fluorescence diminished by about 33% in 80 s, while JC-1 fluorescence diminished by 36% in 200 s. Our results show the loss of Ca
2+
from mitochondria when ΔΨ
m
is depolarised and demonstrate the usefulness of CaOr-5N to mark mitochondrial [Ca
2+
]
m
.</description><subject>Aldehydes</subject><subject>Animals</subject><subject>Benzimidazoles</subject><subject>Biomedical and Life Sciences</subject><subject>Biomedicine</subject><subject>Calcium</subject><subject>Calcium - metabolism</subject><subject>Carbocyanines</subject><subject>Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone - pharmacology</subject><subject>Cell Biology</subject><subject>Dyes</subject><subject>Fluorescent Dyes</subject><subject>Human Physiology</subject><subject>Membrane Potential, Mitochondrial - drug effects</subject><subject>Mice</subject><subject>Microscopy, Confocal</subject><subject>Mitochondria</subject><subject>Mitochondria, Muscle - metabolism</subject><subject>Mitochondria, Muscle - ultrastructure</subject><subject>Molecular Medicine</subject><subject>Muscle Fibers, Skeletal - metabolism</subject><subject>Muscle Fibers, Skeletal - ultrastructure</subject><subject>Muscle, Skeletal - metabolism</subject><subject>Muscle, Skeletal - ultrastructure</subject><subject>Muscular system</subject><subject>Neurosciences</subject><subject>Organic Chemicals</subject><subject>Proteins</subject><subject>Pyridinium Compounds</subject><subject>Receptors</subject><subject>Sarcomeres - metabolism</subject><subject>Sarcoplasmic Reticulum - metabolism</subject><subject>Skeletal Muscle</subject><subject>Staining and Labeling - methods</subject><subject>Time Factors</subject><subject>Uncoupling Agents - pharmacology</subject><issn>0031-6768</issn><issn>1432-2013</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2008</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>eNp1kU1LAzEQhoMotn78AC8SPHiR6CSbbLZHKX6B2Es9h2x21kb3oybdg__eLC0UBE8ZJs_7zvAOIRccbjmAvosAUkiWSgYZF0wdkCmXmWACeHZIppC6LNd5MSEnMX4CgJCFOCYTrjUokPmU1MsV0iEi7Ws6t43zQ7sItvtApt6ojdTSuEbna-9oa8MXhhFs_aZ3q76rgrdNkokb6jva9qNP_MIGN6ndDtE1SGtfYohn5Ki2TcTz3XtK3h8flvNn9rp4epnfvzKX5WrDZKlnVqsSXGFrWSFK7qqMZwqRo-OgEUthndVY2aJwZZ1VTiaJcpXV2lbZKbne-q5D_z1g3JjWR4dNYztM6xkNHIRQOoFXf8DPfghd2s2IFBvMcg4J4lvIhT7GgLVZB59i-DEczHgBs72AGcvxAkYlzeXOeChbrPaKXeQJEFsgpq8UdNhP_t_1F6_jkTc</recordid><startdate>20080101</startdate><enddate>20080101</enddate><creator>Bolaños, Pura</creator><creator>Guillen, Alis</creator><creator>Rojas, Héctor</creator><creator>Boncompagni, Simona</creator><creator>Caputo, Carlo</creator><general>Springer-Verlag</general><general>Springer Nature B.V</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>3V.</scope><scope>7QP</scope><scope>7TK</scope><scope>7TS</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</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>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>7X8</scope></search><sort><creationdate>20080101</creationdate><title>The use of CalciumOrange-5N as a specific marker of mitochondrial Ca2+ in mouse skeletal muscle fibers</title><author>Bolaños, Pura ; Guillen, Alis ; Rojas, Héctor ; Boncompagni, Simona ; Caputo, Carlo</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c365t-4b79a75b0c8af4dee41cd3135ee1ec107eeb2aca7eda88cbf3dc479a5cda77ad3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2008</creationdate><topic>Aldehydes</topic><topic>Animals</topic><topic>Benzimidazoles</topic><topic>Biomedical and Life Sciences</topic><topic>Biomedicine</topic><topic>Calcium</topic><topic>Calcium - metabolism</topic><topic>Carbocyanines</topic><topic>Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone - pharmacology</topic><topic>Cell Biology</topic><topic>Dyes</topic><topic>Fluorescent Dyes</topic><topic>Human Physiology</topic><topic>Membrane Potential, Mitochondrial - drug effects</topic><topic>Mice</topic><topic>Microscopy, Confocal</topic><topic>Mitochondria</topic><topic>Mitochondria, Muscle - metabolism</topic><topic>Mitochondria, Muscle - ultrastructure</topic><topic>Molecular Medicine</topic><topic>Muscle Fibers, Skeletal - metabolism</topic><topic>Muscle Fibers, Skeletal - ultrastructure</topic><topic>Muscle, Skeletal - metabolism</topic><topic>Muscle, Skeletal - ultrastructure</topic><topic>Muscular system</topic><topic>Neurosciences</topic><topic>Organic Chemicals</topic><topic>Proteins</topic><topic>Pyridinium Compounds</topic><topic>Receptors</topic><topic>Sarcomeres - metabolism</topic><topic>Sarcoplasmic Reticulum - metabolism</topic><topic>Skeletal Muscle</topic><topic>Staining and Labeling - methods</topic><topic>Time Factors</topic><topic>Uncoupling Agents - pharmacology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Bolaños, Pura</creatorcontrib><creatorcontrib>Guillen, Alis</creatorcontrib><creatorcontrib>Rojas, Héctor</creatorcontrib><creatorcontrib>Boncompagni, Simona</creatorcontrib><creatorcontrib>Caputo, Carlo</creatorcontrib><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>Neurosciences Abstracts</collection><collection>Physical Education Index</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</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>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>MEDLINE - Academic</collection><jtitle>Pflügers Archiv</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Bolaños, Pura</au><au>Guillen, Alis</au><au>Rojas, Héctor</au><au>Boncompagni, Simona</au><au>Caputo, Carlo</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>The use of CalciumOrange-5N as a specific marker of mitochondrial Ca2+ in mouse skeletal muscle fibers</atitle><jtitle>Pflügers Archiv</jtitle><stitle>Pflugers Arch - Eur J Physiol</stitle><addtitle>Pflugers Arch</addtitle><date>2008-01-01</date><risdate>2008</risdate><volume>455</volume><issue>4</issue><spage>721</spage><epage>731</epage><pages>721-731</pages><issn>0031-6768</issn><eissn>1432-2013</eissn><abstract>We report the use of the fluorescent dye CalciumOrange-5N (CaOr-5N) as a specific mitochondria Ca
2+
marker in enzymatically dissociated mouse FBD muscle fibers. Using laser scanning confocal microscopy and the dyes Mitotracker Green (MTG), di-8-ANEPPS and endoplasmic reticulum tracker green (ERTG), we determined the relative position of mitochondria, transverse tubules and sarcoplasmic reticulum in the sarcomere. Comparison with electron micrographies showed that mitochondria are mostly present at both sides of Z lines and near the triads located at the A-I band border. CaOr-5N fluorescence was mainly distributed in mitochondria, highly co-localised with MTG and basically excluded from the A band space. ERTG localised mostly between the two t-tubules present in each sarcomere. We studied the effect of the protonophore FCCP using CaOr-5N to measure mitochondrial Ca
2+
and JC-1 dye to measure mitochondria inner membrane potential (Δ
Ψ
m
). After FCCP treatment, the CaOr-5N fluorescence diminished by about 33% in 80 s, while JC-1 fluorescence diminished by 36% in 200 s. Our results show the loss of Ca
2+
from mitochondria when ΔΨ
m
is depolarised and demonstrate the usefulness of CaOr-5N to mark mitochondrial [Ca
2+
]
m
.</abstract><cop>Berlin/Heidelberg</cop><pub>Springer-Verlag</pub><pmid>17705046</pmid><doi>10.1007/s00424-007-0312-5</doi><tpages>11</tpages></addata></record> |
| fulltext | fulltext |
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| ispartof | Pflügers Archiv, 2008-01, Vol.455 (4), p.721-731 |
| issn | 0031-6768 1432-2013 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_70102257 |
| source | MEDLINE; Springer Nature - Complete Springer Journals |
| subjects | Aldehydes Animals Benzimidazoles Biomedical and Life Sciences Biomedicine Calcium Calcium - metabolism Carbocyanines Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone - pharmacology Cell Biology Dyes Fluorescent Dyes Human Physiology Membrane Potential, Mitochondrial - drug effects Mice Microscopy, Confocal Mitochondria Mitochondria, Muscle - metabolism Mitochondria, Muscle - ultrastructure Molecular Medicine Muscle Fibers, Skeletal - metabolism Muscle Fibers, Skeletal - ultrastructure Muscle, Skeletal - metabolism Muscle, Skeletal - ultrastructure Muscular system Neurosciences Organic Chemicals Proteins Pyridinium Compounds Receptors Sarcomeres - metabolism Sarcoplasmic Reticulum - metabolism Skeletal Muscle Staining and Labeling - methods Time Factors Uncoupling Agents - pharmacology |
| title | The use of CalciumOrange-5N as a specific marker of mitochondrial Ca2+ in mouse skeletal muscle fibers |
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