Bactericidal antibiotics increase hydroxyphenyl fluorescein signal by altering cell morphology
It was recently proposed that for bactericidal antibiotics a common killing mechanism contributes to lethality involving indirect stimulation of hydroxyl radical (OH•) formation. Flow cytometric detection of OH• by hydroxyphenyl fluorescein (HPF) probe oxidation was used to support this hypothesis....
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| Veröffentlicht in: | PloS one 2014-03, Vol.9 (3), p.e92231-e92231 |
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| creator | Paulander, Wilhelm Wang, Ying Folkesson, Anders Charbon, Godefroid Løbner-Olesen, Anders Ingmer, Hanne |
| description | It was recently proposed that for bactericidal antibiotics a common killing mechanism contributes to lethality involving indirect stimulation of hydroxyl radical (OH•) formation. Flow cytometric detection of OH• by hydroxyphenyl fluorescein (HPF) probe oxidation was used to support this hypothesis. Here we show that increased HPF signals in antibiotics-exposed bacterial cells are explained by fluorescence associated with increased cell size, and do not reflect reactive oxygen species (ROS) concentration. Independently of antibiotics, increased fluorescence was seen for elongated cells expressing the oxidative insensitive green fluorescent protein (GFP). Although our data question the role of ROS in lethality of antibiotics other research approaches point to important interplays between basic bacterial metabolism and antibiotic susceptibility. To underpin such relationships, methods for detecting bacterial metabolites at a cellular level are needed. |
| doi_str_mv | 10.1371/journal.pone.0092231 |
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Flow cytometric detection of OH• by hydroxyphenyl fluorescein (HPF) probe oxidation was used to support this hypothesis. Here we show that increased HPF signals in antibiotics-exposed bacterial cells are explained by fluorescence associated with increased cell size, and do not reflect reactive oxygen species (ROS) concentration. Independently of antibiotics, increased fluorescence was seen for elongated cells expressing the oxidative insensitive green fluorescent protein (GFP). Although our data question the role of ROS in lethality of antibiotics other research approaches point to important interplays between basic bacterial metabolism and antibiotic susceptibility. To underpin such relationships, methods for detecting bacterial metabolites at a cellular level are needed.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0092231</identifier><identifier>PMID: 24647480</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Anti-Bacterial Agents - pharmacology ; Antibiotics ; Apoptosis ; Bacteria ; Biology ; Biology and Life Sciences ; Cell division ; Cell morphology ; Cell size ; Cytology ; E coli ; Elongation ; Escherichia coli ; Escherichia coli - cytology ; Escherichia coli - drug effects ; Flow Cytometry ; Fluorescein ; Fluorescein - metabolism ; Fluoresceins - metabolism ; Fluorescence ; Green fluorescent protein ; Green Fluorescent Proteins - metabolism ; Hydroxyl radicals ; Hypotheses ; Killing ; Lethality ; Metabolism ; Metabolites ; Microscopy ; Microscopy, Interference ; Morphology ; Oxidation ; Oxygen ; Physiological aspects ; Proteins ; Reactive oxygen species ; Research and Analysis Methods ; Science</subject><ispartof>PloS one, 2014-03, Vol.9 (3), p.e92231-e92231</ispartof><rights>COPYRIGHT 2014 Public Library of Science</rights><rights>2014 Paulander et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2014 Paulander et al 2014 Paulander et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c692t-545db9e47e503788657d850ee98db25b4650874557d666d223b4ea67e55805ad3</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3960231/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3960231/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,860,881,2095,2914,23846,27903,27904,53769,53771,79346,79347</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24647480$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Marinus, Martin G.</contributor><creatorcontrib>Paulander, Wilhelm</creatorcontrib><creatorcontrib>Wang, Ying</creatorcontrib><creatorcontrib>Folkesson, Anders</creatorcontrib><creatorcontrib>Charbon, Godefroid</creatorcontrib><creatorcontrib>Løbner-Olesen, Anders</creatorcontrib><creatorcontrib>Ingmer, Hanne</creatorcontrib><title>Bactericidal antibiotics increase hydroxyphenyl fluorescein signal by altering cell morphology</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>It was recently proposed that for bactericidal antibiotics a common killing mechanism contributes to lethality involving indirect stimulation of hydroxyl radical (OH•) formation. Flow cytometric detection of OH• by hydroxyphenyl fluorescein (HPF) probe oxidation was used to support this hypothesis. Here we show that increased HPF signals in antibiotics-exposed bacterial cells are explained by fluorescence associated with increased cell size, and do not reflect reactive oxygen species (ROS) concentration. Independently of antibiotics, increased fluorescence was seen for elongated cells expressing the oxidative insensitive green fluorescent protein (GFP). Although our data question the role of ROS in lethality of antibiotics other research approaches point to important interplays between basic bacterial metabolism and antibiotic susceptibility. 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Flow cytometric detection of OH• by hydroxyphenyl fluorescein (HPF) probe oxidation was used to support this hypothesis. Here we show that increased HPF signals in antibiotics-exposed bacterial cells are explained by fluorescence associated with increased cell size, and do not reflect reactive oxygen species (ROS) concentration. Independently of antibiotics, increased fluorescence was seen for elongated cells expressing the oxidative insensitive green fluorescent protein (GFP). Although our data question the role of ROS in lethality of antibiotics other research approaches point to important interplays between basic bacterial metabolism and antibiotic susceptibility. To underpin such relationships, methods for detecting bacterial metabolites at a cellular level are needed.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>24647480</pmid><doi>10.1371/journal.pone.0092231</doi><tpages>e92231</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Anti-Bacterial Agents - pharmacology Antibiotics Apoptosis Bacteria Biology Biology and Life Sciences Cell division Cell morphology Cell size Cytology E coli Elongation Escherichia coli Escherichia coli - cytology Escherichia coli - drug effects Flow Cytometry Fluorescein Fluorescein - metabolism Fluoresceins - metabolism Fluorescence Green fluorescent protein Green Fluorescent Proteins - metabolism Hydroxyl radicals Hypotheses Killing Lethality Metabolism Metabolites Microscopy Microscopy, Interference Morphology Oxidation Oxygen Physiological aspects Proteins Reactive oxygen species Research and Analysis Methods Science |
| title | Bactericidal antibiotics increase hydroxyphenyl fluorescein signal by altering cell morphology |
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