Estimation of the electric plasma membrane potential difference in yeast with fluorescent dyes: comparative study of methods
Different methods to estimate the plasma membrane potential difference (PMP) of yeast cells with fluorescent monitors were compared. The validity of the methods was tested by the fluorescence difference with or without glucose, and its decrease by the addition of 10 mM KCl. Low CaCl₂ concentrations...
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| Veröffentlicht in: | Journal of bioenergetics and biomembranes 2010-10, Vol.42 (5), p.419-432 |
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| creator | Peña, Antonio Sánchez, Norma Silvia Calahorra, Martha |
| description | Different methods to estimate the plasma membrane potential difference (PMP) of yeast cells with fluorescent monitors were compared. The validity of the methods was tested by the fluorescence difference with or without glucose, and its decrease by the addition of 10 mM KCl. Low CaCl₂ concentrations avoid binding of the dye to the cell surface, and low CCCP concentrations avoid its accumulation by mitochondria. Lower concentrations of Ba²⁺ produce a similar effect as Ca²⁺, without producing the fluorescence changes derived from its transport. Fluorescence changes without considering binding of the dyes to the cells and accumulation by mitochondria are overshadowed by their distribution between this organelle and the cytoplasm. Other factors, such as yeast starvation, dye used, parameters of the fluorescence changes, as well as buffers and incubation times were analyzed. An additional approach to measure the actual or relative values of PMP, determining the accumulation of the dye, is presented. |
| doi_str_mv | 10.1007/s10863-010-9311-x |
| format | Article |
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The validity of the methods was tested by the fluorescence difference with or without glucose, and its decrease by the addition of 10 mM KCl. Low CaCl₂ concentrations avoid binding of the dye to the cell surface, and low CCCP concentrations avoid its accumulation by mitochondria. Lower concentrations of Ba²⁺ produce a similar effect as Ca²⁺, without producing the fluorescence changes derived from its transport. Fluorescence changes without considering binding of the dyes to the cells and accumulation by mitochondria are overshadowed by their distribution between this organelle and the cytoplasm. Other factors, such as yeast starvation, dye used, parameters of the fluorescence changes, as well as buffers and incubation times were analyzed. An additional approach to measure the actual or relative values of PMP, determining the accumulation of the dye, is presented.</description><identifier>ISSN: 0145-479X</identifier><identifier>EISSN: 1573-6881</identifier><identifier>DOI: 10.1007/s10863-010-9311-x</identifier><identifier>PMID: 21063758</identifier><language>eng</language><publisher>Boston: Boston : Springer US</publisher><subject>Accumulation ; Animal Anatomy ; Animal Biochemistry ; Biochemistry ; Biological Transport, Active - physiology ; Bioorganic Chemistry ; Calcium - metabolism ; Calcium Chloride - metabolism ; Carbonyl Cyanide m-Chlorophenyl Hydrazone - metabolism ; Cell Membrane - physiology ; Cell Respiration - physiology ; Chemistry ; Chemistry and Materials Science ; Comparative studies ; Dyes ; Fluorescence ; Fluorescent Dyes ; Histology ; ion transport ; Membrane potential indicators ; Membrane Potentials - physiology ; Membranes ; Microscopy, Confocal ; Morphology ; Organic Chemistry ; Plasma ; Potassium - metabolism ; Potassium Chloride - metabolism ; Saccharomyces cerevisiae - physiology ; Yeast ; Yeasts</subject><ispartof>Journal of bioenergetics and biomembranes, 2010-10, Vol.42 (5), p.419-432</ispartof><rights>Springer Science+Business Media, LLC 2010</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c426t-7a94fff3c88c41b6997cedd382a35f0b76a1cc416c66ad34ac51b52bca5525453</citedby><cites>FETCH-LOGICAL-c426t-7a94fff3c88c41b6997cedd382a35f0b76a1cc416c66ad34ac51b52bca5525453</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/s10863-010-9311-x$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s10863-010-9311-x$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>315,781,785,27929,27930,41493,42562,51324</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21063758$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Peña, Antonio</creatorcontrib><creatorcontrib>Sánchez, Norma Silvia</creatorcontrib><creatorcontrib>Calahorra, Martha</creatorcontrib><title>Estimation of the electric plasma membrane potential difference in yeast with fluorescent dyes: comparative study of methods</title><title>Journal of bioenergetics and biomembranes</title><addtitle>J Bioenerg Biomembr</addtitle><addtitle>J Bioenerg Biomembr</addtitle><description>Different methods to estimate the plasma membrane potential difference (PMP) of yeast cells with fluorescent monitors were compared. The validity of the methods was tested by the fluorescence difference with or without glucose, and its decrease by the addition of 10 mM KCl. Low CaCl₂ concentrations avoid binding of the dye to the cell surface, and low CCCP concentrations avoid its accumulation by mitochondria. Lower concentrations of Ba²⁺ produce a similar effect as Ca²⁺, without producing the fluorescence changes derived from its transport. Fluorescence changes without considering binding of the dyes to the cells and accumulation by mitochondria are overshadowed by their distribution between this organelle and the cytoplasm. Other factors, such as yeast starvation, dye used, parameters of the fluorescence changes, as well as buffers and incubation times were analyzed. An additional approach to measure the actual or relative values of PMP, determining the accumulation of the dye, is presented.</description><subject>Accumulation</subject><subject>Animal Anatomy</subject><subject>Animal Biochemistry</subject><subject>Biochemistry</subject><subject>Biological Transport, Active - physiology</subject><subject>Bioorganic Chemistry</subject><subject>Calcium - metabolism</subject><subject>Calcium Chloride - metabolism</subject><subject>Carbonyl Cyanide m-Chlorophenyl Hydrazone - metabolism</subject><subject>Cell Membrane - physiology</subject><subject>Cell Respiration - physiology</subject><subject>Chemistry</subject><subject>Chemistry and Materials Science</subject><subject>Comparative studies</subject><subject>Dyes</subject><subject>Fluorescence</subject><subject>Fluorescent Dyes</subject><subject>Histology</subject><subject>ion transport</subject><subject>Membrane potential indicators</subject><subject>Membrane Potentials - physiology</subject><subject>Membranes</subject><subject>Microscopy, Confocal</subject><subject>Morphology</subject><subject>Organic Chemistry</subject><subject>Plasma</subject><subject>Potassium - metabolism</subject><subject>Potassium Chloride - metabolism</subject><subject>Saccharomyces cerevisiae - physiology</subject><subject>Yeast</subject><subject>Yeasts</subject><issn>0145-479X</issn><issn>1573-6881</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</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>eNqFkUtv1TAQhSMEopfCD2ADFhtWgZn4EZtdVZWHVIkFVGJnOY7dmyqJg-1Ar8SPx1cpILGAlRfznXM8c6rqKcIrBGhfJwQpaA0ItaKI9e29aoe8pbWQEu9XO0DGa9aqLyfVo5RuAEACh4fVSYMgaMvlrvpxkfIwmTyEmQRP8t4RNzqb42DJMpo0GTK5qYtmdmQJ2c15MCPpB-9ddLN1ZJjJwZmUyfch74kf1xBdsoUj_cGlN8SGaTGxBHxzJOW1PxxjJpf3oU-PqwfejMk9uXtPq6u3F5_P39eXH999OD-7rC1rRK5bo5j3nlopLcNOKNVa1_dUNoZyD10rDNoyEVYI01NmLMeON501nDeccXpavdx8lxi-ri5lPQ3lj-NYtgpr0golMCo4_JeUyJhoFGeFfPEXeRPWOJc1tASFTAHKAuEG2RhSis7rJZZrx4NG0McK9VahLhXqY4X6tmie3Rmv3eT634pfnRWg2YBURvO1i3-S_-X6fBN5E7S5jkPSV58aQAqokDKq6E8DFrI2</recordid><startdate>20101001</startdate><enddate>20101001</enddate><creator>Peña, Antonio</creator><creator>Sánchez, Norma Silvia</creator><creator>Calahorra, Martha</creator><general>Boston : Springer US</general><general>Springer US</general><general>Springer Nature B.V</general><scope>FBQ</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>3V.</scope><scope>7QO</scope><scope>7QP</scope><scope>7TK</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>88I</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7P</scope><scope>P64</scope><scope>PDBOC</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>7X8</scope><scope>M7N</scope></search><sort><creationdate>20101001</creationdate><title>Estimation of the electric plasma membrane potential difference in yeast with fluorescent dyes: comparative study of methods</title><author>Peña, Antonio ; Sánchez, Norma Silvia ; Calahorra, Martha</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c426t-7a94fff3c88c41b6997cedd382a35f0b76a1cc416c66ad34ac51b52bca5525453</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>Accumulation</topic><topic>Animal Anatomy</topic><topic>Animal Biochemistry</topic><topic>Biochemistry</topic><topic>Biological Transport, Active - physiology</topic><topic>Bioorganic Chemistry</topic><topic>Calcium - metabolism</topic><topic>Calcium Chloride - metabolism</topic><topic>Carbonyl Cyanide m-Chlorophenyl Hydrazone - metabolism</topic><topic>Cell Membrane - physiology</topic><topic>Cell Respiration - physiology</topic><topic>Chemistry</topic><topic>Chemistry and Materials Science</topic><topic>Comparative studies</topic><topic>Dyes</topic><topic>Fluorescence</topic><topic>Fluorescent Dyes</topic><topic>Histology</topic><topic>ion transport</topic><topic>Membrane potential indicators</topic><topic>Membrane Potentials - physiology</topic><topic>Membranes</topic><topic>Microscopy, Confocal</topic><topic>Morphology</topic><topic>Organic Chemistry</topic><topic>Plasma</topic><topic>Potassium - metabolism</topic><topic>Potassium Chloride - metabolism</topic><topic>Saccharomyces cerevisiae - physiology</topic><topic>Yeast</topic><topic>Yeasts</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Peña, Antonio</creatorcontrib><creatorcontrib>Sánchez, Norma Silvia</creatorcontrib><creatorcontrib>Calahorra, Martha</creatorcontrib><collection>AGRIS</collection><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>Biotechnology Research Abstracts</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Neurosciences Abstracts</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>Science Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology 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>Materials Science & Engineering Collection</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>Technology Collection (ProQuest)</collection><collection>Natural Science Collection (ProQuest)</collection><collection>ProQuest One Community College</collection><collection>ProQuest Materials Science Collection</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</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>Materials Science Database</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Science Database (ProQuest)</collection><collection>Biological Science Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Materials Science Collection</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central Basic</collection><collection>MEDLINE - Academic</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><jtitle>Journal of bioenergetics and biomembranes</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Peña, Antonio</au><au>Sánchez, Norma Silvia</au><au>Calahorra, Martha</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Estimation of the electric plasma membrane potential difference in yeast with fluorescent dyes: comparative study of methods</atitle><jtitle>Journal of bioenergetics and biomembranes</jtitle><stitle>J Bioenerg Biomembr</stitle><addtitle>J Bioenerg Biomembr</addtitle><date>2010-10-01</date><risdate>2010</risdate><volume>42</volume><issue>5</issue><spage>419</spage><epage>432</epage><pages>419-432</pages><issn>0145-479X</issn><eissn>1573-6881</eissn><abstract>Different methods to estimate the plasma membrane potential difference (PMP) of yeast cells with fluorescent monitors were compared. The validity of the methods was tested by the fluorescence difference with or without glucose, and its decrease by the addition of 10 mM KCl. Low CaCl₂ concentrations avoid binding of the dye to the cell surface, and low CCCP concentrations avoid its accumulation by mitochondria. Lower concentrations of Ba²⁺ produce a similar effect as Ca²⁺, without producing the fluorescence changes derived from its transport. Fluorescence changes without considering binding of the dyes to the cells and accumulation by mitochondria are overshadowed by their distribution between this organelle and the cytoplasm. Other factors, such as yeast starvation, dye used, parameters of the fluorescence changes, as well as buffers and incubation times were analyzed. An additional approach to measure the actual or relative values of PMP, determining the accumulation of the dye, is presented.</abstract><cop>Boston</cop><pub>Boston : Springer US</pub><pmid>21063758</pmid><doi>10.1007/s10863-010-9311-x</doi><tpages>14</tpages></addata></record> |
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| subjects | Accumulation Animal Anatomy Animal Biochemistry Biochemistry Biological Transport, Active - physiology Bioorganic Chemistry Calcium - metabolism Calcium Chloride - metabolism Carbonyl Cyanide m-Chlorophenyl Hydrazone - metabolism Cell Membrane - physiology Cell Respiration - physiology Chemistry Chemistry and Materials Science Comparative studies Dyes Fluorescence Fluorescent Dyes Histology ion transport Membrane potential indicators Membrane Potentials - physiology Membranes Microscopy, Confocal Morphology Organic Chemistry Plasma Potassium - metabolism Potassium Chloride - metabolism Saccharomyces cerevisiae - physiology Yeast Yeasts |
| title | Estimation of the electric plasma membrane potential difference in yeast with fluorescent dyes: comparative study of methods |
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