A High-Affinity Fluorescent Sensor for Catecholamine: Application to Monitoring Norepinephrine Exocytosis.

A fluorescent sensor for catecholamines, NS510, is presented. The sensor is based on a quinolone fluorophore incorporating a boronic acid recognition element that gives it high affinity for catecholamines and a turn-on response to norepinephrine. The sensor results in punctate staining of norepinephrine-enriched chromaffin cells visualized using confocal microscopy indicating that it stains the norepinephrine in secretory vesicles. Amperometry in conjunction with total internal reflection fluorescence (TIRF) microscopy demonstrates that the sensor can be used to observe destaining of individual chromaffin granules upon exocytosis. NS510 is the highest affinity fluorescent norepinephrine sensor currently available and can be used for measuring catecholamines in live-cell assays.

[1]  Sylvie Maurin,et al.  A Dual Functional Electroactive and Fluorescent Probe for Coupled Measurements of Vesicular Exocytosis with High Spatial and Temporal Resolution. , 2017, Angewandte Chemie.

[2]  D. Sulzer,et al.  Fluorescent false neurotransmitter reveals functionally silent dopamine vesicle clusters in the striatum , 2016, Nature Neuroscience.

[3]  Kenneth S Hettie,et al.  Turn-On Near-Infrared Fluorescent Sensor for Selectively Imaging Serotonin. , 2016, ACS chemical neuroscience.

[4]  G. Shi,et al.  Development of Au Disk Nanoelectrode Down to 3 nm in Radius for Detection of Dopamine Release from a Single Cell. , 2015, Analytical chemistry.

[5]  Kenneth S Hettie,et al.  Coumarin-3-aldehyde as a scaffold for the design of tunable PET-modulated fluorescent sensors for neurotransmitters. , 2014, Chemistry.

[6]  K. Gillis,et al.  Selective catecholamine recognition with NeuroSensor 521: a fluorescent sensor for the visualization of norepinephrine in fixed and live cells. , 2013, ACS chemical neuroscience.

[7]  C. Barnard,et al.  Fluorescent dopamine tracer resolves individual dopaminergic synapses and their activity in the brain , 2012, Proceedings of the National Academy of Sciences.

[8]  D. V. van Essen,et al.  Challenges and Opportunities in Mining Neuroscience Data , 2011, Science.

[9]  K. Olson,et al.  Neurotransmitters excreted in the urine as biomarkers of nervous system activity: Validity and clinical applicability , 2011, Neuroscience & Biobehavioral Reviews.

[10]  D. Sulzer,et al.  Development of pH-responsive fluorescent false neurotransmitters. , 2010, Journal of the American Chemical Society.

[11]  Christy L Haynes,et al.  Bioanalytical tools for single-cell study of exocytosis , 2010, Analytical and bioanalytical chemistry.

[12]  P. Vallotton,et al.  Exocytotic Vesicle Behaviour Assessed by Total Internal Reflection Fluorescence Microscopy , 2010, Traffic.

[13]  Andrew G Ewing,et al.  Only a Fraction of Quantal Content is Released During Exocytosis as Revealed by Electrochemical Cytometry of Secretory Vesicles. , 2010, ACS chemical neuroscience.

[14]  D. Sulzer,et al.  Fluorescent False Neurotransmitters Visualize Dopamine Release from Individual Presynaptic Terminals , 2009, Science.

[15]  T. Wolff,et al.  Lewis‐acid‐catalyzed Photodimerization of Coumarins and N‐methyl‐2‐quinolone , 2008, Photochemistry and photobiology.

[16]  Hong Wang,et al.  Determination of amino acid neurotransmitters in human cerebrospinal fluid and saliva by capillary electrophoresis with laser-induced fluorescence detection. , 2008, Journal of separation science.

[17]  V. Tran,et al.  Computer‐Based De Novo Design, Synthesis, and Evaluation of Boronic Acid‐Based Artificial Receptors for Selective Recognition of Dopamine , 2008, Chembiochem : a European journal of chemical biology.

[18]  K. Gillis,et al.  Phosphomimetic Mutation of Ser-187 of SNAP-25 Increases both Syntaxin Binding and Highly Ca2+-sensitive Exocytosis , 2007, The Journal of general physiology.

[19]  T. Glass,et al.  Fluorescent sensors for diamines , 2005 .

[20]  T. Schrader,et al.  A color sensor for catecholamines. , 2005, Angewandte Chemie.

[21]  T. Glass,et al.  Selective amine recognition: development of a chemosensor for dopamine and norepinephrine. , 2004, Organic letters.

[22]  R. Kennedy,et al.  Trace-level amino acid analysis by capillary liquid chromatography and application to in vivo microdialysis sampling with 10-s temporal resolution. , 2000, Analytical chemistry.

[23]  R N Zare,et al.  Patch-Clamp Detection of Neurotransmitters in Capillary Electrophoresis , 1996, Science.

[24]  J. A. Jankowski,et al.  Temporally resolved catecholamine spikes correspond to single vesicle release from individual chromaffin cells. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[25]  M. A. Moro,et al.  Separation and culture of living adrenaline- and noradrenaline-containing cells from bovine adrenal medullae. , 1990, Analytical biochemistry.

[26]  K. Gillis,et al.  Electrochemical measurement of quantal exocytosis using microchips , 2017, Pflügers Archiv - European Journal of Physiology.

[27]  D. O'Connor,et al.  Human dopamine β-hydroxylase promoter variant alters transcription in chromaffin cells, enzyme secretion, and blood pressure. , 2011, American journal of hypertension.

[28]  Juyoung Yoon,et al.  Fluorescence Sensing of Dopamine , 2005 .