Patulin-induced cellular toxicity: A vital fluorescence study
The mechanisms of patulin-induced cellular toxicity in an immortalized rat granulosa cell line were examined using several vital fluorescence bioassays. Monochlorobimane and 5-chloromethylfluorescein diacetate were used to monitor cellular glutathione (GSH) levels and revealed dose- and time-depende...
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| Veröffentlicht in: | Toxicology and applied pharmacology 1992-02, Vol.112 (2), p.235-244 |
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| creator | Burghardt, Robert C. Barhoumi, Rola Lewis, Erik H. Bailey, R.Hartford Pyle, Kristen A. Clement, Beverly A. Phillips, Timothy D. |
| description | The mechanisms of patulin-induced cellular toxicity in an immortalized rat granulosa cell line were examined using several vital fluorescence bioassays. Monochlorobimane and 5-chloromethylfluorescein diacetate were used to monitor cellular glutathione (GSH) levels and revealed dose- and time-dependent depletion of GSH by patulin. A significant reduction in the fluorescence of the monochlorobimane-GSH conjugate by 0.1 μ
m patulin was observed between 1 and 2 hr. Similar GSH depletion by the mycotoxin was also observed in parallel studies on a liver (Clone 9) and a renal (LLC-PK
1) cell line, although reduction of fluorescence occurred within 1 hr at the same dosage. Analysis of the electrical potential-dependent partitioning of rhodamine 123 into mitochondria also revealed significant effects of patulin within 1 hr at 0.1 μ
m. An initial dose-dependent reduction in mitochondrial fluorescence was followed by loss of selective partitioning of the fluorophore into mitochondria at higher doses and/or a longer exposure of cells to patulin. The reduction in mitochondrial fluorescence was paralleled by a dose-dependent decrease in intracellular pH detected with 2′,7′-bis-(2-carboxyethyl)-5(6)-carboxyfluorescein. Analysis of [Ca
2+]
i with indo-1 and fluo-3 revealed a significant dose-dependent influx of Ca
2+ at 10 μ
m and an alteration of the pattern of ionomycin-induced Ca
2+ influx at 1.0 μ
m following patulin treatment. A carboxy-fluorescein fluorescence photobleaching assay was used to examine the effects of patulin on gap junction-mediated intercellular communication. Dose-dependent reduction in intercellular communication was observed within 2 hr with 1.0 μ
m patulin. These observations indicate that the fluorescence assays used in this study provide a sensitive index of toxicity caused by exposure to patulin. Further, the toxic effects of patulin may involve direct effects on cellular glutathione levels and mitochondrial function in addition to direct effects on the plasma membrane. |
| doi_str_mv | 10.1016/0041-008X(92)90193-V |
| format | Article |
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m patulin was observed between 1 and 2 hr. Similar GSH depletion by the mycotoxin was also observed in parallel studies on a liver (Clone 9) and a renal (LLC-PK
1) cell line, although reduction of fluorescence occurred within 1 hr at the same dosage. Analysis of the electrical potential-dependent partitioning of rhodamine 123 into mitochondria also revealed significant effects of patulin within 1 hr at 0.1 μ
m. An initial dose-dependent reduction in mitochondrial fluorescence was followed by loss of selective partitioning of the fluorophore into mitochondria at higher doses and/or a longer exposure of cells to patulin. The reduction in mitochondrial fluorescence was paralleled by a dose-dependent decrease in intracellular pH detected with 2′,7′-bis-(2-carboxyethyl)-5(6)-carboxyfluorescein. Analysis of [Ca
2+]
i with indo-1 and fluo-3 revealed a significant dose-dependent influx of Ca
2+ at 10 μ
m and an alteration of the pattern of ionomycin-induced Ca
2+ influx at 1.0 μ
m following patulin treatment. A carboxy-fluorescein fluorescence photobleaching assay was used to examine the effects of patulin on gap junction-mediated intercellular communication. Dose-dependent reduction in intercellular communication was observed within 2 hr with 1.0 μ
m patulin. These observations indicate that the fluorescence assays used in this study provide a sensitive index of toxicity caused by exposure to patulin. Further, the toxic effects of patulin may involve direct effects on cellular glutathione levels and mitochondrial function in addition to direct effects on the plasma membrane.</description><identifier>ISSN: 0041-008X</identifier><identifier>EISSN: 1096-0333</identifier><identifier>DOI: 10.1016/0041-008X(92)90193-V</identifier><identifier>PMID: 1539161</identifier><identifier>CODEN: TXAPA9</identifier><language>eng</language><publisher>San Diego, CA: Elsevier Inc</publisher><subject>Animals ; Biological and medical sciences ; Calcium - metabolism ; Cell Communication - drug effects ; CULTIVO DE CELULAS ; CULTURE DE CELLULE ; DOSAGE BIOLOGIQUE ; ENSAYO BIOLOGICO ; Female ; Fluorescence ; Glutathione - metabolism ; Granulosa Cell Tumor - drug therapy ; Granulosa Cell Tumor - metabolism ; Granulosa Cell Tumor - pathology ; Homeostasis - drug effects ; Hydrogen-Ion Concentration ; Intercellular Junctions - drug effects ; Intracellular Membranes - drug effects ; Kinetics ; Medical sciences ; Membrane Potentials - drug effects ; Mitochondria - drug effects ; Mitochondria - physiology ; Ovarian Neoplasms - drug therapy ; Ovarian Neoplasms - metabolism ; Ovarian Neoplasms - pathology ; Patulin - toxicity ; PATULINA ; PATULINE ; Plant poisons toxicology ; PROPIEDADES OPTICAS ; PROPRIETE OPTIQUE ; Rats ; TOXICIDAD ; TOXICIDAD POR INGESTION ; TOXICITE ; TOXICITE PAR INGESTION ; Toxicology ; Tumor Cells, Cultured - drug effects</subject><ispartof>Toxicology and applied pharmacology, 1992-02, Vol.112 (2), p.235-244</ispartof><rights>1992</rights><rights>1992 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c436t-5e20671712b01191798ca9ad72b36a54d276d14ea7ab87312ea62c339b190bab3</citedby><cites>FETCH-LOGICAL-c436t-5e20671712b01191798ca9ad72b36a54d276d14ea7ab87312ea62c339b190bab3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/0041-008X(92)90193-V$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,778,782,3539,27907,27908,45978</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=5517939$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1539161$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Burghardt, Robert C.</creatorcontrib><creatorcontrib>Barhoumi, Rola</creatorcontrib><creatorcontrib>Lewis, Erik H.</creatorcontrib><creatorcontrib>Bailey, R.Hartford</creatorcontrib><creatorcontrib>Pyle, Kristen A.</creatorcontrib><creatorcontrib>Clement, Beverly A.</creatorcontrib><creatorcontrib>Phillips, Timothy D.</creatorcontrib><title>Patulin-induced cellular toxicity: A vital fluorescence study</title><title>Toxicology and applied pharmacology</title><addtitle>Toxicol Appl Pharmacol</addtitle><description>The mechanisms of patulin-induced cellular toxicity in an immortalized rat granulosa cell line were examined using several vital fluorescence bioassays. Monochlorobimane and 5-chloromethylfluorescein diacetate were used to monitor cellular glutathione (GSH) levels and revealed dose- and time-dependent depletion of GSH by patulin. A significant reduction in the fluorescence of the monochlorobimane-GSH conjugate by 0.1 μ
m patulin was observed between 1 and 2 hr. Similar GSH depletion by the mycotoxin was also observed in parallel studies on a liver (Clone 9) and a renal (LLC-PK
1) cell line, although reduction of fluorescence occurred within 1 hr at the same dosage. Analysis of the electrical potential-dependent partitioning of rhodamine 123 into mitochondria also revealed significant effects of patulin within 1 hr at 0.1 μ
m. An initial dose-dependent reduction in mitochondrial fluorescence was followed by loss of selective partitioning of the fluorophore into mitochondria at higher doses and/or a longer exposure of cells to patulin. The reduction in mitochondrial fluorescence was paralleled by a dose-dependent decrease in intracellular pH detected with 2′,7′-bis-(2-carboxyethyl)-5(6)-carboxyfluorescein. Analysis of [Ca
2+]
i with indo-1 and fluo-3 revealed a significant dose-dependent influx of Ca
2+ at 10 μ
m and an alteration of the pattern of ionomycin-induced Ca
2+ influx at 1.0 μ
m following patulin treatment. A carboxy-fluorescein fluorescence photobleaching assay was used to examine the effects of patulin on gap junction-mediated intercellular communication. Dose-dependent reduction in intercellular communication was observed within 2 hr with 1.0 μ
m patulin. These observations indicate that the fluorescence assays used in this study provide a sensitive index of toxicity caused by exposure to patulin. Further, the toxic effects of patulin may involve direct effects on cellular glutathione levels and mitochondrial function in addition to direct effects on the plasma membrane.</description><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Calcium - metabolism</subject><subject>Cell Communication - drug effects</subject><subject>CULTIVO DE CELULAS</subject><subject>CULTURE DE CELLULE</subject><subject>DOSAGE BIOLOGIQUE</subject><subject>ENSAYO BIOLOGICO</subject><subject>Female</subject><subject>Fluorescence</subject><subject>Glutathione - metabolism</subject><subject>Granulosa Cell Tumor - drug therapy</subject><subject>Granulosa Cell Tumor - metabolism</subject><subject>Granulosa Cell Tumor - pathology</subject><subject>Homeostasis - drug effects</subject><subject>Hydrogen-Ion Concentration</subject><subject>Intercellular Junctions - drug effects</subject><subject>Intracellular Membranes - drug effects</subject><subject>Kinetics</subject><subject>Medical sciences</subject><subject>Membrane Potentials - drug effects</subject><subject>Mitochondria - drug effects</subject><subject>Mitochondria - physiology</subject><subject>Ovarian Neoplasms - drug therapy</subject><subject>Ovarian Neoplasms - metabolism</subject><subject>Ovarian Neoplasms - pathology</subject><subject>Patulin - toxicity</subject><subject>PATULINA</subject><subject>PATULINE</subject><subject>Plant poisons toxicology</subject><subject>PROPIEDADES OPTICAS</subject><subject>PROPRIETE OPTIQUE</subject><subject>Rats</subject><subject>TOXICIDAD</subject><subject>TOXICIDAD POR INGESTION</subject><subject>TOXICITE</subject><subject>TOXICITE PAR INGESTION</subject><subject>Toxicology</subject><subject>Tumor Cells, Cultured - drug effects</subject><issn>0041-008X</issn><issn>1096-0333</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1992</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kNFK3UAQhpei6FH7AqVCLorUi9SZ3WSTLbQgUrUgKLRK75bJ7kZWcpLjbiKetzdpDnrn1Vz83z_MfIx9RviGgPIEIMMUoPz3VfFjBahEeveBLRCUTEEIscUWr8gu24vxAQBUluEO28FcKJS4YD9uqB8a36a-tYNxNjGuaYaGQtJ3z974fv09OU2efE9NUjdDF1w0rjUuif1g1wdsu6Ymuo-buc9uz3_9PbtMr64vfp-dXqUmE7JPc8dBFlggrwBRYaFKQ4pswSshKc8sL6TFzFFBVVkI5I4kN0KoChVUVIl9djTvXYXucXCx10sfp0updd0QNUqOwKEcwWwGTehiDK7Wq-CXFNYaQU_W9KRET0q04vq_NX031g43-4dq6exbadY05l82OUVDTR2oNT6-Ynk-viTUiH2asZo6TfdhRG7_qAyhVGIMf86hG0U9eRd0NH5SaX1wpte28-8f-QKq_4_r</recordid><startdate>19920201</startdate><enddate>19920201</enddate><creator>Burghardt, Robert C.</creator><creator>Barhoumi, Rola</creator><creator>Lewis, Erik H.</creator><creator>Bailey, R.Hartford</creator><creator>Pyle, Kristen A.</creator><creator>Clement, Beverly A.</creator><creator>Phillips, Timothy D.</creator><general>Elsevier Inc</general><general>Elsevier</general><scope>FBQ</scope><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>7U7</scope><scope>C1K</scope><scope>M7N</scope></search><sort><creationdate>19920201</creationdate><title>Patulin-induced cellular toxicity: A vital fluorescence study</title><author>Burghardt, Robert C. ; Barhoumi, Rola ; Lewis, Erik H. ; Bailey, R.Hartford ; Pyle, Kristen A. ; Clement, Beverly A. ; Phillips, Timothy D.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c436t-5e20671712b01191798ca9ad72b36a54d276d14ea7ab87312ea62c339b190bab3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1992</creationdate><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>Calcium - metabolism</topic><topic>Cell Communication - drug effects</topic><topic>CULTIVO DE CELULAS</topic><topic>CULTURE DE CELLULE</topic><topic>DOSAGE BIOLOGIQUE</topic><topic>ENSAYO BIOLOGICO</topic><topic>Female</topic><topic>Fluorescence</topic><topic>Glutathione - metabolism</topic><topic>Granulosa Cell Tumor - drug therapy</topic><topic>Granulosa Cell Tumor - metabolism</topic><topic>Granulosa Cell Tumor - pathology</topic><topic>Homeostasis - drug effects</topic><topic>Hydrogen-Ion Concentration</topic><topic>Intercellular Junctions - drug effects</topic><topic>Intracellular Membranes - drug effects</topic><topic>Kinetics</topic><topic>Medical sciences</topic><topic>Membrane Potentials - drug effects</topic><topic>Mitochondria - drug effects</topic><topic>Mitochondria - physiology</topic><topic>Ovarian Neoplasms - drug therapy</topic><topic>Ovarian Neoplasms - metabolism</topic><topic>Ovarian Neoplasms - pathology</topic><topic>Patulin - toxicity</topic><topic>PATULINA</topic><topic>PATULINE</topic><topic>Plant poisons toxicology</topic><topic>PROPIEDADES OPTICAS</topic><topic>PROPRIETE OPTIQUE</topic><topic>Rats</topic><topic>TOXICIDAD</topic><topic>TOXICIDAD POR INGESTION</topic><topic>TOXICITE</topic><topic>TOXICITE PAR INGESTION</topic><topic>Toxicology</topic><topic>Tumor Cells, Cultured - drug effects</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Burghardt, Robert C.</creatorcontrib><creatorcontrib>Barhoumi, Rola</creatorcontrib><creatorcontrib>Lewis, Erik H.</creatorcontrib><creatorcontrib>Bailey, R.Hartford</creatorcontrib><creatorcontrib>Pyle, Kristen A.</creatorcontrib><creatorcontrib>Clement, Beverly A.</creatorcontrib><creatorcontrib>Phillips, Timothy D.</creatorcontrib><collection>AGRIS</collection><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>Toxicology Abstracts</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><jtitle>Toxicology and applied pharmacology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Burghardt, Robert C.</au><au>Barhoumi, Rola</au><au>Lewis, Erik H.</au><au>Bailey, R.Hartford</au><au>Pyle, Kristen A.</au><au>Clement, Beverly A.</au><au>Phillips, Timothy D.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Patulin-induced cellular toxicity: A vital fluorescence study</atitle><jtitle>Toxicology and applied pharmacology</jtitle><addtitle>Toxicol Appl Pharmacol</addtitle><date>1992-02-01</date><risdate>1992</risdate><volume>112</volume><issue>2</issue><spage>235</spage><epage>244</epage><pages>235-244</pages><issn>0041-008X</issn><eissn>1096-0333</eissn><coden>TXAPA9</coden><abstract>The mechanisms of patulin-induced cellular toxicity in an immortalized rat granulosa cell line were examined using several vital fluorescence bioassays. Monochlorobimane and 5-chloromethylfluorescein diacetate were used to monitor cellular glutathione (GSH) levels and revealed dose- and time-dependent depletion of GSH by patulin. A significant reduction in the fluorescence of the monochlorobimane-GSH conjugate by 0.1 μ
m patulin was observed between 1 and 2 hr. Similar GSH depletion by the mycotoxin was also observed in parallel studies on a liver (Clone 9) and a renal (LLC-PK
1) cell line, although reduction of fluorescence occurred within 1 hr at the same dosage. Analysis of the electrical potential-dependent partitioning of rhodamine 123 into mitochondria also revealed significant effects of patulin within 1 hr at 0.1 μ
m. An initial dose-dependent reduction in mitochondrial fluorescence was followed by loss of selective partitioning of the fluorophore into mitochondria at higher doses and/or a longer exposure of cells to patulin. The reduction in mitochondrial fluorescence was paralleled by a dose-dependent decrease in intracellular pH detected with 2′,7′-bis-(2-carboxyethyl)-5(6)-carboxyfluorescein. Analysis of [Ca
2+]
i with indo-1 and fluo-3 revealed a significant dose-dependent influx of Ca
2+ at 10 μ
m and an alteration of the pattern of ionomycin-induced Ca
2+ influx at 1.0 μ
m following patulin treatment. A carboxy-fluorescein fluorescence photobleaching assay was used to examine the effects of patulin on gap junction-mediated intercellular communication. Dose-dependent reduction in intercellular communication was observed within 2 hr with 1.0 μ
m patulin. These observations indicate that the fluorescence assays used in this study provide a sensitive index of toxicity caused by exposure to patulin. Further, the toxic effects of patulin may involve direct effects on cellular glutathione levels and mitochondrial function in addition to direct effects on the plasma membrane.</abstract><cop>San Diego, CA</cop><pub>Elsevier Inc</pub><pmid>1539161</pmid><doi>10.1016/0041-008X(92)90193-V</doi><tpages>10</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0041-008X |
| ispartof | Toxicology and applied pharmacology, 1992-02, Vol.112 (2), p.235-244 |
| issn | 0041-008X 1096-0333 |
| language | eng |
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| source | MEDLINE; Elsevier ScienceDirect Journals |
| subjects | Animals Biological and medical sciences Calcium - metabolism Cell Communication - drug effects CULTIVO DE CELULAS CULTURE DE CELLULE DOSAGE BIOLOGIQUE ENSAYO BIOLOGICO Female Fluorescence Glutathione - metabolism Granulosa Cell Tumor - drug therapy Granulosa Cell Tumor - metabolism Granulosa Cell Tumor - pathology Homeostasis - drug effects Hydrogen-Ion Concentration Intercellular Junctions - drug effects Intracellular Membranes - drug effects Kinetics Medical sciences Membrane Potentials - drug effects Mitochondria - drug effects Mitochondria - physiology Ovarian Neoplasms - drug therapy Ovarian Neoplasms - metabolism Ovarian Neoplasms - pathology Patulin - toxicity PATULINA PATULINE Plant poisons toxicology PROPIEDADES OPTICAS PROPRIETE OPTIQUE Rats TOXICIDAD TOXICIDAD POR INGESTION TOXICITE TOXICITE PAR INGESTION Toxicology Tumor Cells, Cultured - drug effects |
| title | Patulin-induced cellular toxicity: A vital fluorescence study |
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