Abstract
AbstractA 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 turnāon response to norepinephrine. The sensor results in punctate staining of norepinephrineāenriched 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 liveācell assays.
🔬 Techniques
🔭 Microscopes
💻 Software
✨ Fluorophores
🏭 Microscope Brands
📷 Detectors
🔎 Objectives
💻 Software Details
🏛️ Research Organizations (ROR)
Affiliated research institutions:
📊 Figures
Figure 1.
Binding of two catecholamine sensors to norepinephrine
Figure 2.
(a) UV/vis titration and (b) fluorescence titration of the sensor NS510 (10 u00b5M in 25 mM HEPES, 50 mM Na 2 S 2 O 3 , pH = 7.4) with norepinephrine (10 mM in buffer, u03bb ex = 488 nm). Inset in (b)...
Figure 3.
Confocal image of norepinephrine-enriched cells incubated with NS510 (0.5 u00b5M) (u03bb ex = 488 nm). (Scale bar = 10 u00b5m)
Figure 4.
Combination of amperometry with TIRF imaging demonstrates that NS510 can resolve catecholamine release from an individual vesicle. Three recordings from three different bovine adrenal chromaffin cells...
Scheme 1.
Synthesis of NS510
Figure images are served from the NIH/NLM PubMed Central Open Access Subset or Europe PMC; copyright remains with the publishers and authors.
💬 Discussion
0 commentsNo comments yet. Be the first to start a discussion!
Leave a Comment