Shedding light on the D1-like receptors: a fluorescence-based toolbox for visualization of the D1 and D5 receptors

Niklas Rosier, Denise Mönnich, Martin Nagl, Hannes Schihada, Alexei Sirbu, Nergis Konar, Irene Reyes-Resina, Gemma Navarro, Rafael Franco, Peter Kolb, Paolo Annibale, Steffen Pockes*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

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Abstract

Dopamine D1-like receptors are the most abundant type of dopamine receptors in the central nervous system and, even after decades of discovery, still highly interesting for the study of neurological diseases. We herein describe the synthesis of a new set of fluorescent ligands, structurally derived from D1R antagonist SCH-23390 and labeled with two different fluorescent dyes, as tool compounds for the visualization of D1-like receptors. Pharmacological characterization in radioligand binding studies identified UR-NR435 (25) as a high-affinity ligand for D1-like receptors (pKi (D1R) = 8.34, pKi (D5R) = 7.62) with excellent selectivity towards D2-like receptors. Compound 25 proved to be a neutral antagonist at the D1R and D5R in a Gs heterotrimer dissociation assay, an important feature to avoid receptor internalization and degradation when working with whole cells. The neutral antagonist 25 displayed rapid association and complete dissociation to the D1R in kinetic binding studies using confocal microscopy verifying its applicability for fluorescence microscopy. Moreover, molecular brightness studies determined a single-digit nanomolar binding affinity of the ligand, which was in good agreement with radioligand binding data. For this reason, this fluorescent ligand is a useful tool for a sophisticated characterization of native D1 receptors in a variety of experimental setups.
Original languageEnglish
Article numbere202300658
JournalChemBioChem
VolumeEarly View
Early online date20 Nov 2023
DOIs
Publication statusE-pub ahead of print - 20 Nov 2023

Keywords

  • Confocal microscopy
  • D1-like receptors
  • Dopamine receptors
  • Fluorescent ligands
  • Molecular brightness

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