A Genetically Encoded Ratiometric Indicator for Chloride: Capturing Chloride Transients in Cultured Hippocampal Neurons
We constructed a novel optical indicator for chloride ions by fusing the chloride-sensitive yellow fluorescent protein with the chloride-insensitive cyan fluorescent protein. The ratio of FRET-dependent emission of these fluorophores varied in proportion to the concentration of Cl − and was used to...
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| Veröffentlicht in: | Neuron (Cambridge, Mass.) Mass.), 2000-09, Vol.27 (3), p.447-459 |
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| creator | Kuner, Thomas Augustine, George J. |
| description | We constructed a novel optical indicator for chloride ions by fusing the chloride-sensitive yellow fluorescent protein with the chloride-insensitive cyan fluorescent protein. The ratio of FRET-dependent emission of these fluorophores varied in proportion to the concentration of Cl
− and was used to measure intracellular chloride concentration ([Cl
−]
i) in cultured hippocampal neurons. [Cl
−]
i decreased during neuronal development, consistent with the shift from excitation to inhibition during maturation of GABAergic synapses. Focal activation of GABA
A receptors caused large changes in [Cl
−]
i that could underlie use-dependent depression of GABA-dependent synaptic transmission. GABA-induced changes in somatic [Cl
−]
i spread into dendrites, suggesting that [Cl
−]
i can signal the location of synaptic activity. This genetically encoded indicator will permit new approaches ranging from high-throughput drug screening to direct recordings of synaptic Cl
− signals in vivo. |
| doi_str_mv | 10.1016/S0896-6273(00)00056-8 |
| format | Article |
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− and was used to measure intracellular chloride concentration ([Cl
−]
i) in cultured hippocampal neurons. [Cl
−]
i decreased during neuronal development, consistent with the shift from excitation to inhibition during maturation of GABAergic synapses. Focal activation of GABA
A receptors caused large changes in [Cl
−]
i that could underlie use-dependent depression of GABA-dependent synaptic transmission. GABA-induced changes in somatic [Cl
−]
i spread into dendrites, suggesting that [Cl
−]
i can signal the location of synaptic activity. This genetically encoded indicator will permit new approaches ranging from high-throughput drug screening to direct recordings of synaptic Cl
− signals in vivo.</description><identifier>ISSN: 0896-6273</identifier><identifier>EISSN: 1097-4199</identifier><identifier>DOI: 10.1016/S0896-6273(00)00056-8</identifier><identifier>PMID: 11055428</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Animals ; Bacterial Proteins - genetics ; Bacterial Proteins - metabolism ; Cells, Cultured ; Chlorides - analysis ; Chlorides - metabolism ; Dendrites - metabolism ; Energy Transfer ; Fluorescent Dyes ; gamma-Aminobutyric Acid - metabolism ; Hippocampus - cytology ; Hippocampus - metabolism ; Hydrogen-Ion Concentration ; Intracellular Fluid - chemistry ; Intracellular Fluid - metabolism ; Luminescent Proteins - genetics ; Luminescent Proteins - metabolism ; Neurons - cytology ; Neurons - metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, GABA-A - metabolism ; Recombinant Fusion Proteins - genetics ; Recombinant Fusion Proteins - metabolism ; Signal Transduction - physiology ; Spectrometry, Fluorescence ; Synapses - metabolism ; Synaptic Transmission - physiology ; Transfection</subject><ispartof>Neuron (Cambridge, Mass.), 2000-09, Vol.27 (3), p.447-459</ispartof><rights>2000 Cell Press</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c346t-2a0117b9c85fd9dacf992d81ad5d3c45f4bf9807683255d16de20cce8ec5ade13</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/S0896-6273(00)00056-8$$EHTML$$P50$$Gelsevier$$Hfree_for_read</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11055428$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kuner, Thomas</creatorcontrib><creatorcontrib>Augustine, George J.</creatorcontrib><title>A Genetically Encoded Ratiometric Indicator for Chloride: Capturing Chloride Transients in Cultured Hippocampal Neurons</title><title>Neuron (Cambridge, Mass.)</title><addtitle>Neuron</addtitle><description>We constructed a novel optical indicator for chloride ions by fusing the chloride-sensitive yellow fluorescent protein with the chloride-insensitive cyan fluorescent protein. The ratio of FRET-dependent emission of these fluorophores varied in proportion to the concentration of Cl
− and was used to measure intracellular chloride concentration ([Cl
−]
i) in cultured hippocampal neurons. [Cl
−]
i decreased during neuronal development, consistent with the shift from excitation to inhibition during maturation of GABAergic synapses. Focal activation of GABA
A receptors caused large changes in [Cl
−]
i that could underlie use-dependent depression of GABA-dependent synaptic transmission. GABA-induced changes in somatic [Cl
−]
i spread into dendrites, suggesting that [Cl
−]
i can signal the location of synaptic activity. This genetically encoded indicator will permit new approaches ranging from high-throughput drug screening to direct recordings of synaptic Cl
− signals in vivo.</description><subject>Animals</subject><subject>Bacterial Proteins - genetics</subject><subject>Bacterial Proteins - metabolism</subject><subject>Cells, Cultured</subject><subject>Chlorides - analysis</subject><subject>Chlorides - metabolism</subject><subject>Dendrites - metabolism</subject><subject>Energy Transfer</subject><subject>Fluorescent Dyes</subject><subject>gamma-Aminobutyric Acid - metabolism</subject><subject>Hippocampus - cytology</subject><subject>Hippocampus - metabolism</subject><subject>Hydrogen-Ion Concentration</subject><subject>Intracellular Fluid - chemistry</subject><subject>Intracellular Fluid - metabolism</subject><subject>Luminescent Proteins - genetics</subject><subject>Luminescent Proteins - metabolism</subject><subject>Neurons - cytology</subject><subject>Neurons - metabolism</subject><subject>Rats</subject><subject>Rats, Sprague-Dawley</subject><subject>Receptors, GABA-A - metabolism</subject><subject>Recombinant Fusion Proteins - genetics</subject><subject>Recombinant Fusion Proteins - metabolism</subject><subject>Signal Transduction - physiology</subject><subject>Spectrometry, Fluorescence</subject><subject>Synapses - metabolism</subject><subject>Synaptic Transmission - physiology</subject><subject>Transfection</subject><issn>0896-6273</issn><issn>1097-4199</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkUFv1DAQha0K1C6lP6HIJwSHwDiJHZtLtYpKW6kCCcrZ8tqT1iixg-2A-u_JtqVXDqORZr550rxHyCmDDwyY-PgdpBKVqLvmHcB7AOCikgdkw0B1VcuUekE2z8gReZXzTwDWcsUOyRFjwHlbyw35s6UXGLB4a8bxnp4HGx06-s0UHycsyVt6Fdy6LTHRYa3-bozJO_xEezOXJflw-zyjN8mE7DGUTH2g_TKuwKp26ec5WjPNZqRfcEkx5Nfk5WDGjCdP_Zj8-Hx-019W118vrvrtdWWbVpSqNsBYt1NW8sEpZ-ygVO0kM467xrZ8aHeDktAJ2dScOyYc1mAtSrTcOGTNMXn7qDun-GvBXPTks8VxNAHjknVXN_tb9V-QdaKVAGIF3zyBy25Cp-fkJ5Pu9T9PV-DsEcD1r98ek8529cSi8wlt0S56zUDvU9QPKep9RBpAP6SoZfMXQ0WP2Q</recordid><startdate>20000901</startdate><enddate>20000901</enddate><creator>Kuner, Thomas</creator><creator>Augustine, George J.</creator><general>Elsevier Inc</general><scope>6I.</scope><scope>AAFTH</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7TK</scope><scope>7X8</scope></search><sort><creationdate>20000901</creationdate><title>A Genetically Encoded Ratiometric Indicator for Chloride: Capturing Chloride Transients in Cultured Hippocampal Neurons</title><author>Kuner, Thomas ; Augustine, George J.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c346t-2a0117b9c85fd9dacf992d81ad5d3c45f4bf9807683255d16de20cce8ec5ade13</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2000</creationdate><topic>Animals</topic><topic>Bacterial Proteins - genetics</topic><topic>Bacterial Proteins - metabolism</topic><topic>Cells, Cultured</topic><topic>Chlorides - analysis</topic><topic>Chlorides - metabolism</topic><topic>Dendrites - metabolism</topic><topic>Energy Transfer</topic><topic>Fluorescent Dyes</topic><topic>gamma-Aminobutyric Acid - metabolism</topic><topic>Hippocampus - cytology</topic><topic>Hippocampus - metabolism</topic><topic>Hydrogen-Ion Concentration</topic><topic>Intracellular Fluid - chemistry</topic><topic>Intracellular Fluid - metabolism</topic><topic>Luminescent Proteins - genetics</topic><topic>Luminescent Proteins - metabolism</topic><topic>Neurons - cytology</topic><topic>Neurons - metabolism</topic><topic>Rats</topic><topic>Rats, Sprague-Dawley</topic><topic>Receptors, GABA-A - metabolism</topic><topic>Recombinant Fusion Proteins - genetics</topic><topic>Recombinant Fusion Proteins - metabolism</topic><topic>Signal Transduction - physiology</topic><topic>Spectrometry, Fluorescence</topic><topic>Synapses - metabolism</topic><topic>Synaptic Transmission - physiology</topic><topic>Transfection</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kuner, Thomas</creatorcontrib><creatorcontrib>Augustine, George J.</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>Neurosciences Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Neuron (Cambridge, Mass.)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kuner, Thomas</au><au>Augustine, George J.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A Genetically Encoded Ratiometric Indicator for Chloride: Capturing Chloride Transients in Cultured Hippocampal Neurons</atitle><jtitle>Neuron (Cambridge, Mass.)</jtitle><addtitle>Neuron</addtitle><date>2000-09-01</date><risdate>2000</risdate><volume>27</volume><issue>3</issue><spage>447</spage><epage>459</epage><pages>447-459</pages><issn>0896-6273</issn><eissn>1097-4199</eissn><abstract>We constructed a novel optical indicator for chloride ions by fusing the chloride-sensitive yellow fluorescent protein with the chloride-insensitive cyan fluorescent protein. The ratio of FRET-dependent emission of these fluorophores varied in proportion to the concentration of Cl
− and was used to measure intracellular chloride concentration ([Cl
−]
i) in cultured hippocampal neurons. [Cl
−]
i decreased during neuronal development, consistent with the shift from excitation to inhibition during maturation of GABAergic synapses. Focal activation of GABA
A receptors caused large changes in [Cl
−]
i that could underlie use-dependent depression of GABA-dependent synaptic transmission. GABA-induced changes in somatic [Cl
−]
i spread into dendrites, suggesting that [Cl
−]
i can signal the location of synaptic activity. This genetically encoded indicator will permit new approaches ranging from high-throughput drug screening to direct recordings of synaptic Cl
− signals in vivo.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>11055428</pmid><doi>10.1016/S0896-6273(00)00056-8</doi><tpages>13</tpages><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; Cell Press Free Archives; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; ScienceDirect Journals (5 years ago - present) |
| subjects | Animals Bacterial Proteins - genetics Bacterial Proteins - metabolism Cells, Cultured Chlorides - analysis Chlorides - metabolism Dendrites - metabolism Energy Transfer Fluorescent Dyes gamma-Aminobutyric Acid - metabolism Hippocampus - cytology Hippocampus - metabolism Hydrogen-Ion Concentration Intracellular Fluid - chemistry Intracellular Fluid - metabolism Luminescent Proteins - genetics Luminescent Proteins - metabolism Neurons - cytology Neurons - metabolism Rats Rats, Sprague-Dawley Receptors, GABA-A - metabolism Recombinant Fusion Proteins - genetics Recombinant Fusion Proteins - metabolism Signal Transduction - physiology Spectrometry, Fluorescence Synapses - metabolism Synaptic Transmission - physiology Transfection |
| title | A Genetically Encoded Ratiometric Indicator for Chloride: Capturing Chloride Transients in Cultured Hippocampal Neurons |
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