Synthesis and Characterization of a Novel Concentration-Independent Fluorescent Chloride Indicator, ABP-Dextran, Optimized for Extracellular Chloride Measurement
GABA, the primary inhibitory neurotransmitter, stimulates GABAA receptors (GABAARs) to increase the chloride conductance of the cytosolic membrane. The driving forces for membrane chloride currents are determined by the local differences between intracellular and extracellular chloride concentration...
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| Format: | Article |
| Language: | English |
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MDPI AG
2024-01-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/14/1/77 |
| _version_ | 1797344567390896128 |
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| author | Kieran P. Normoyle Kyle P. Lillis Kevin J. Staley |
| author_facet | Kieran P. Normoyle Kyle P. Lillis Kevin J. Staley |
| author_sort | Kieran P. Normoyle |
| collection | DOAJ |
| description | GABA, the primary inhibitory neurotransmitter, stimulates GABAA receptors (GABAARs) to increase the chloride conductance of the cytosolic membrane. The driving forces for membrane chloride currents are determined by the local differences between intracellular and extracellular chloride concentrations (Cli and Clo, respectively). While several strategies exist for the measurement of Cli, the field lacks tools for non-invasive measurement of Clo. We present the design and development of a <b>f</b>luorescent <b>l</b>ifetime <b>im</b>aging (FLIM)-compatible small molecule, N(4-<b>a</b>mino<b>b</b>utyl)<b>p</b>henanthridiunium (ABP) with the brightness, spectral features, sensitivity to chloride, and selectivity versus other anions to serve as a useful probe of Clo. ABP can be conjugated to dextran to ensure extracellular compartmentalization, and a second chloride-insensitive counter-label can be added for ratiometric imaging. We validate the utility of this novel sensor series in two sensor concentration-independent modes: FLIM or ratiometric intensity-based imaging. |
| first_indexed | 2024-03-08T11:04:35Z |
| format | Article |
| id | doaj.art-a68879ae5f58441d972bfa05f88743dc |
| institution | Directory Open Access Journal |
| issn | 2218-273X |
| language | English |
| last_indexed | 2024-03-08T11:04:35Z |
| publishDate | 2024-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomolecules |
| spelling | doaj.art-a68879ae5f58441d972bfa05f88743dc2024-01-26T15:19:28ZengMDPI AGBiomolecules2218-273X2024-01-011417710.3390/biom14010077Synthesis and Characterization of a Novel Concentration-Independent Fluorescent Chloride Indicator, ABP-Dextran, Optimized for Extracellular Chloride MeasurementKieran P. Normoyle0Kyle P. Lillis1Kevin J. Staley2Massachusetts General Hospital, Department of Neurology, 55 Fruit Street, Boston, MA 02114, USAMassachusetts General Hospital, Department of Neurology, 55 Fruit Street, Boston, MA 02114, USAMassachusetts General Hospital, Department of Neurology, 55 Fruit Street, Boston, MA 02114, USAGABA, the primary inhibitory neurotransmitter, stimulates GABAA receptors (GABAARs) to increase the chloride conductance of the cytosolic membrane. The driving forces for membrane chloride currents are determined by the local differences between intracellular and extracellular chloride concentrations (Cli and Clo, respectively). While several strategies exist for the measurement of Cli, the field lacks tools for non-invasive measurement of Clo. We present the design and development of a <b>f</b>luorescent <b>l</b>ifetime <b>im</b>aging (FLIM)-compatible small molecule, N(4-<b>a</b>mino<b>b</b>utyl)<b>p</b>henanthridiunium (ABP) with the brightness, spectral features, sensitivity to chloride, and selectivity versus other anions to serve as a useful probe of Clo. ABP can be conjugated to dextran to ensure extracellular compartmentalization, and a second chloride-insensitive counter-label can be added for ratiometric imaging. We validate the utility of this novel sensor series in two sensor concentration-independent modes: FLIM or ratiometric intensity-based imaging.https://www.mdpi.com/2218-273X/14/1/77extracellular chlorideextracellular matrixfluorescent chloride sensorfluorescence lifetime imagingfluorescent dextran |
| spellingShingle | Kieran P. Normoyle Kyle P. Lillis Kevin J. Staley Synthesis and Characterization of a Novel Concentration-Independent Fluorescent Chloride Indicator, ABP-Dextran, Optimized for Extracellular Chloride Measurement Biomolecules extracellular chloride extracellular matrix fluorescent chloride sensor fluorescence lifetime imaging fluorescent dextran |
| title | Synthesis and Characterization of a Novel Concentration-Independent Fluorescent Chloride Indicator, ABP-Dextran, Optimized for Extracellular Chloride Measurement |
| title_full | Synthesis and Characterization of a Novel Concentration-Independent Fluorescent Chloride Indicator, ABP-Dextran, Optimized for Extracellular Chloride Measurement |
| title_fullStr | Synthesis and Characterization of a Novel Concentration-Independent Fluorescent Chloride Indicator, ABP-Dextran, Optimized for Extracellular Chloride Measurement |
| title_full_unstemmed | Synthesis and Characterization of a Novel Concentration-Independent Fluorescent Chloride Indicator, ABP-Dextran, Optimized for Extracellular Chloride Measurement |
| title_short | Synthesis and Characterization of a Novel Concentration-Independent Fluorescent Chloride Indicator, ABP-Dextran, Optimized for Extracellular Chloride Measurement |
| title_sort | synthesis and characterization of a novel concentration independent fluorescent chloride indicator abp dextran optimized for extracellular chloride measurement |
| topic | extracellular chloride extracellular matrix fluorescent chloride sensor fluorescence lifetime imaging fluorescent dextran |
| url | https://www.mdpi.com/2218-273X/14/1/77 |
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