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Free, ionic zinc (Zn2+) modulates neurotransmitter dynamics in the brain. However, the sub-s effects of transient concentration changes of Zn2+ on neurotransmitter release and uptake are not well understood. To address this lack of knowledge, we have combined the photolysis of the novel caged Zn2+ compound [Zn(DPAdeCageOMe)]+ with fast scan cyclic voltammetry (FSCV) at carbon fiber microelectrodes in live, whole brain preparations from zebrafish (Danio rerio). After treating the brain with [Zn(DPAdeCageOMe)]+, Zn2+ was released by application of light that was gated through a computer-controlled shutter synchronized with the FSCV measurements and delivered through a 1 mm fiber optic cable. We systematically optimized the photocage concentration and light application parameters, including the total duration and light-to-electrical stimulation delay time. While sub-s Zn2+ application with this method inhibited DA reuptake, assessed by the first-order rate constant (k) and half-life (t1/2), it had no effect on the electrically stimulated DA overflow ([DA]STIM). Increasing the photocage concentration and light duration progressively inhibited uptake, with maximal effects occurring at 100 μM and 800 ms, respectively. Furthermore, uptake was inhibited 200 ms after Zn2+ photorelease, but no measurable effect occurred after 800 ms. We expect that application of this method to the zebrafish whole brain and other preparations will help expand the current knowledge of how Zn2+ affects neurotransmitter release/uptake in select neurological disease states. 

Abstract MorphDeCage (2‐(4‐methoxy‐3‐nitrophenyl)‐2‐morpholinoacetic acid) and PyrDeCage (2‐(4‐methoxy‐3‐nitrophenyl)‐2‐(methyl(pyridin‐2‐ylmethyl)amino)acetic) are Zn²⁺photocages that utilize photodecarboxylation of the methoxy derivative ofmeta‐nitrophenylacetic acid as the release mechanism. Isothermal titration calorimetry (ITC) was used an alternative to usual approaches to measure the Zn²⁺binding affinities of these new compounds owing to unsuccessful measurement by competitive titration with 4‐(2‐pyridylazo)resorcinol (PAR). MorphDeCage forms a 1 : 1 ligand‐metal complex with a 106 μM K_dvalue. PyrDeCage forms both a 1 : 1 and 1 : 2 metal: ligand complexes with 3.2 and 21.7 μM K_dvalues respectively. To further demonstrate the efficacy of the ITC methodology and provide a comparison to direct UV‐vis titrations data, two photocages based on Sanger's reagent (SRPs) were prepared. The K_dvalues of the SRPs measured by UV‐vis titration and ITC were internally consistent and support the retraction of the original report (J. Am. Chem. Soc.2020,142, 3806–3813), which was withdrawn due to errors in binding affinity measurements.