Informace o publikaci

Toward Serotonin Fluorescent False Neurotransmitters: Development of Fluorescent Dual Serotonin and Vesicular Monoamine Transporter Substrates for Visualizing Serotonin Neurons

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HENKE Adam KOVALYOVA Yekaterina DUNN Matthew DREIER Dominik GUBERNATOR Niko G. DINCHEVA Iva HWU Christopher ŠEBEJ Peter ANSORGE Mark S. SULZER David SAMEŠ Dalibor

Druh Článek v odborném periodiku
Časopis / Zdroj ACS CHEMICAL NEUROSCIENCE
Citace
www https://pubs.acs.org/doi/abs/10.1021/acschemneuro.7b00320
Doi http://dx.doi.org/10.1021/acschemneuro.7b00320
Klíčová slova FFN246; fluorescent false neurotransmitters; serotonin; 5-HT; monoaminergic neurons; imaging probe
Popis Ongoing efforts in our laboratories focus on design of optical reporters known as fluorescent false neurotransmitters (FFNs) that enable the visualization of uptake into, packaging within, and release from individual monoaminergic neurons and presynaptic sites in the brain. Here, we introduce the molecular probe FFN246 as an expansion of the FFN platform to the serotonergic system. Combining the acridone fluorophore with the ethylamine recognition element of serotonin, we identified FFN54 and FFN246 as substrates for both the serotonin transporter and the vesicular monoamine transporter 2 (VMAT2). A systematic structure-activity study revealed the basic structural chemotype of aminoalkyl acridones required for serotonin transporter (SERT) activity and enabled lowering the background labeling of these probes while maintaining SERT activity, which proved essential for obtaining sufficient signal in the brain tissue (FFN246). We demonstrate the utility of FFN246 for direct examination of SERT activity and SERT inhibitors in 96-well cell culture assays, as well as specific labeling of serotonergic neurons of the dorsal raphe nucleus in the living tissue of acute mouse brain slices. While we found only minor FFN246 accumulation in serotonergic axons in murine brain tissue, FFN246 effectively traces serotonin uptake and packaging in the soma of serotonergic neurons with improved photophysical properties and loading parameters compared to known serotonin-based fluorescent tracers.