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Article: Analysis of mitochondria by capillary electrophoresis: Cardiolipin levels decrease in response to carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone

TitleAnalysis of mitochondria by capillary electrophoresis: Cardiolipin levels decrease in response to carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone
Authors
KeywordsCapillary electrophoresis
Cardiolipin
FCCP
Mitochondria
Issue Date2010
PublisherWiley - V C H Verlag GmbH & Co KGaA. The Journal's web site is located at http://www3.interscience.wiley.com/cgi-bin/jhome/69502350
Citation
European Journal Of Lipid Science And Technology, 2010, v. 112 n. 9, p. 1058-1066 How to Cite?
AbstractCardiolipin is an important phospholipid present in the inner membrane of mitochondria. It plays a critical role in adenosine triphosphate (ATP) synthesis mediated by oxidative phosphorylation. Exposure of HepG2 cells to carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP) caused the inhibition of ATP synthesis and the depolarization of mitochondria. Capillary electrophoresis with laser-induced fluorescence (CE-LIF) analysis of fluorescent mitochondrion-selective probe 10-N-nonyl acridine orange (NAO) labeled mitochondria was employed to in situ estimate the cardiolipin levels under FCCP-induced de-energization of mitochondria. NAO, stoichiometriclly bound to cardiolipin at a 1:1 or 2:1 molar ratio (NAO/cardiolipin), emitted green and red fluorescence, respectively. Green fluorescence was chosen for cardiolipin content analysis because it was more intense than red fluorescence. A significant decrease in the cardiolipin content, up to 11% of the control, was evident when the ATP content and mitochondrial membrane potential (MMP) correspondingly decreased. These related findings suggested that CE-LIF may provide a sensitive strategy to determine cardiolipin content in response to exposure to chemical uncouplers. This reinforces the hypothesis that alterations in ATP synthesis and MMP have a close association with cardiolipin content, which correlated tightly with mitochondrial membrane assembly and activity. © 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Persistent Identifierhttp://hdl.handle.net/10722/124494
ISSN
2015 Impact Factor: 1.953
2015 SCImago Journal Rankings: 0.654
ISI Accession Number ID
Funding AgencyGrant Number
Hong Kong Research Grants Council of the Hong Kong Special Administrative Region, ChinaHKU7023/07P
Hong Kong University Research and Conference Grants Committee
Funding Information:

We would like to thank Dr. Wilson Ching Yickpang for kindly providing HepG2 cells and the support from the Hong Kong Research Grants Council of the Hong Kong Special Administrative Region, China (HKU7023/07P) and the Hong Kong University Research and Conference Grants Committee.

References

 

DC FieldValueLanguage
dc.contributor.authorZhao, Wen_HK
dc.contributor.authorO, Wen_HK
dc.contributor.authorFung, Yen_HK
dc.contributor.authorCheung, MPen_HK
dc.date.accessioned2010-10-31T10:37:33Z-
dc.date.available2010-10-31T10:37:33Z-
dc.date.issued2010en_HK
dc.identifier.citationEuropean Journal Of Lipid Science And Technology, 2010, v. 112 n. 9, p. 1058-1066en_HK
dc.identifier.issn1438-7697en_HK
dc.identifier.urihttp://hdl.handle.net/10722/124494-
dc.description.abstractCardiolipin is an important phospholipid present in the inner membrane of mitochondria. It plays a critical role in adenosine triphosphate (ATP) synthesis mediated by oxidative phosphorylation. Exposure of HepG2 cells to carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP) caused the inhibition of ATP synthesis and the depolarization of mitochondria. Capillary electrophoresis with laser-induced fluorescence (CE-LIF) analysis of fluorescent mitochondrion-selective probe 10-N-nonyl acridine orange (NAO) labeled mitochondria was employed to in situ estimate the cardiolipin levels under FCCP-induced de-energization of mitochondria. NAO, stoichiometriclly bound to cardiolipin at a 1:1 or 2:1 molar ratio (NAO/cardiolipin), emitted green and red fluorescence, respectively. Green fluorescence was chosen for cardiolipin content analysis because it was more intense than red fluorescence. A significant decrease in the cardiolipin content, up to 11% of the control, was evident when the ATP content and mitochondrial membrane potential (MMP) correspondingly decreased. These related findings suggested that CE-LIF may provide a sensitive strategy to determine cardiolipin content in response to exposure to chemical uncouplers. This reinforces the hypothesis that alterations in ATP synthesis and MMP have a close association with cardiolipin content, which correlated tightly with mitochondrial membrane assembly and activity. © 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.en_HK
dc.languageengen_HK
dc.publisherWiley - V C H Verlag GmbH & Co KGaA. The Journal's web site is located at http://www3.interscience.wiley.com/cgi-bin/jhome/69502350en_HK
dc.relation.ispartofEuropean Journal of Lipid Science and Technologyen_HK
dc.subjectCapillary electrophoresisen_HK
dc.subjectCardiolipinen_HK
dc.subjectFCCPen_HK
dc.subjectMitochondriaen_HK
dc.titleAnalysis of mitochondria by capillary electrophoresis: Cardiolipin levels decrease in response to carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazoneen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=1438-7697&volume=112&spage=1058&epage=1066&date=2010&atitle=Analysis+of+mitochondria+by+capillary+electrophoresis:+Cardiolipin+levels+decrease+in+response+to+carbonyl+cyanide+4-(trifluoromethoxy)+phenylhydrazoneen_HK
dc.identifier.emailO, W:owaisum@hkucc.hku.hken_HK
dc.identifier.emailFung, Y:ysfung@hku.hken_HK
dc.identifier.authorityO, W=rp00315en_HK
dc.identifier.authorityFung, Y=rp00697en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1002/ejlt.201000018en_HK
dc.identifier.scopuseid_2-s2.0-77956905976en_HK
dc.identifier.hkuros182875en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-77956905976&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume112en_HK
dc.identifier.issue9en_HK
dc.identifier.spage1058en_HK
dc.identifier.epage1066en_HK
dc.identifier.isiWOS:000282662800016-
dc.publisher.placeGermanyen_HK
dc.identifier.scopusauthoridZhao, W=34871080300en_HK
dc.identifier.scopusauthoridO, W=6701729369en_HK
dc.identifier.scopusauthoridFung, Y=13309754700en_HK
dc.identifier.scopusauthoridCheung, MP=16749051200en_HK

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