A Dual Functional Electroactive and Fluorescent Probe for Coupled Measurements of Vesicular Exocytosis with High Spatial and Temporal Resolution. Issue 9 (24th January 2017)

• Record Type: Journal Article
• Authors: Liu, Xiaoqing; Savy, Alexandra; Maurin, Sylvie; Grimaud, Laurence; Darchen, François; Quinton, Damien; Labbé, Eric; Buriez, Olivier; Delacotte, Jérôme; Lemaître, Frédéric; Guille‐Collignon, Manon
• Is Part Of: Angewandte Chemie international edition. Volume 56:Issue 9(2017)
• Page Start: 2366
• Page End: 2370
• Publication Date: 2017-01-24
• Subjects: electrochemistry -- exocytosis -- indium tin oxides -- neurotransmitters; Chemistry -- Periodicals; 540
• DOI: 10.1002/anie.201611145
• Languages: English
• ISSNs: 1433-7851

Abstract

Abstract: In this work, Fluorescent False Neurotransmitter 102 (FFN102), a synthesized analogue of biogenic neurotransmitters, was demonstrated to show both pH‐dependent fluorescence and electroactivity. To study secretory behaviors at the single‐vesicle level, FFN102 was employed as a new fluorescent/electroactive dual probe in a coupled technique (amperometry and total internal reflection fluorescence microscopy (TIRFM)). We used N13 cells, a stable clone of BON cells, to specifically accumulate FFN102 into their secretory vesicles, and then optical and electrochemical measurements of vesicular exocytosis were experimentally achieved by using indium tin oxide (ITO) transparent electrodes. Upon stimulation, FFN102 started to diffuse out from the acidic intravesicular microenvironment to the neutral extracellular space, leading to fluorescent emissions and to the electrochemical oxidation signals that were simultaneously collected from the ITO electrode surface. The correlation of fluorescence and amperometric signals resulting from the FFN102 probe allows real‐time monitoring of single exocytotic events with both high spatial and temporal resolution. This work opens new possibilities in the investigation of exocytotic mechanisms. Abstract : The synthetic neurotransmitter FFN102 is employed to monitor coupled amperometric and fluorescence microscopy measurements of individual exocytotic events. This method capitalizes on the strengths of both individual techniques, and opens the way for new approaches to study the kinetics and mechanisms of vesicular secretion.