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Substrate induced currents through the monoamine transporters and single cell uptake of the fluorescent substrate APP+

Citation

Sandtner, Walter (2021), Substrate induced currents through the monoamine transporters and single cell uptake of the fluorescent substrate APP+, Dryad, Dataset, https://doi.org/10.5061/dryad.6q573n5z8

Abstract

The concentrative power of the transporters for dopamine (DAT), norepinephrine (NET) and serotonin (SERT) is thought to be fueled by the transmembrane Na+ gradient, but it is conceivable that they can also tap other energy sources, e.g. membrane voltage and/or the transmembrane K+ gradient. We address this by recording uptake of endogenous substrates or the fluorescent substrate APP+ ((4-(4-dimethylamino)phenyl-1-methylpyridinium) under voltage control in cells expressing DAT, NET or SERT. We show that DAT and NET differ from SERT in intracellular handling of K+. In DAT and NET, substrate uptake was voltage-dependent due to the transient nature of intracellular K+ binding, which precluded K+ antiport. SERT, however, antiports K+ and achieves voltage-independent transport. Thus, there is a trade-off between maintaining constant uptake and harvesting membrane potential for concentrative power, which we conclude to occur due to subtle differences in the kinetics of co-substrate ion binding in closely related transporters.

Methods

Collected via Clampex (Axon-Instruments) . It has not been processed. The data sets are raw files.

Usage Notes

Can be visualized and analyzed with Clampfit (Axon-Instruments).

Funding

Austrian Science Fund, Award: P 31813