RT Journal Article
SR Electronic
T1 Pharmacological characterization of the zebrafish (<em>Danio rerio</em>) histamine H<sub>1</sub> receptor reveals the involvement of the second extracellular loop in the binding of histamine
JF Molecular Pharmacology
JO Mol Pharmacol
FD American Society for Pharmacology and Experimental Therapeutics
SP MOLPHARM-AR-2023-000741
DO 10.1124/molpharm.123.000741
A1 Daniel A McNaught-Flores
A1 Albert J Kooistra
A1 Yu-Chia Chen
A1 Jose-Antonio Arias-Montano
A1 Pertti Panula
A1 Rob Leurs
YR 2023
UL http://molpharm.aspetjournals.org/content/early/2023/11/17/molpharm.123.000741.abstract
AB The zebrafish (Danio rerio) histamine H1 receptor gene (zfH1R) was cloned in 2007 and reported to be involved in fish locomotion. Yet, no detailed characterization of its pharmacology and signaling properties have so far been reported. In this study, we pharmacologically characterized the zfH1R expressed in HEK-293T cells by means of [3H]-mepyramine binding and G protein signaling assays. The zfH1R (KD 0.7 nM) displayed similar affinity for the antagonist [3H]-mepyramine as the hH1R (KD 1.5 nM), whereas the affinity for histamine is 100-fold higher than for the human H1R. The zfH1R couples to Gaq/11 proteins and activates several reporter genes, i.e. NFAT, NFkB, CRE, VEGF, COX-2, SRE and AP-1, and zfH1R-mediated signaling is prevented by the Gaq/11 inhibitor YM-254890 and the antagonist mepyramine. Molecular modeling of the zfH1R and human H1R shows that the binding pockets are identical, implying that variations along the ligand binding pathway could underly the differences in histamine affinity instead. Targeting differentially charged residues in extracellular loop 2 (ECL2) using site-directed mutagenesis revealed that Arg21045x55 is most likely involved in the binding process of histamine in zfH1R. This study aids the understanding of the pharmacological differences between H1R orthologs and the role of ECL2 in histamine binding and provides fundamental information for the understanding of the histaminergic system in the zebrafish. Significance Statement The use of the zebrafish as in vivo model in neuroscience is growing exponentially, which asks for detailed characterization of the aminergic neurotransmitter systems in this model. This study is the first to pharmacologically characterize the zebrafish histamine H1 receptor after expression in HEK-293T cells. Our results show a high pharmacological and functional resemblance with the human ortholog but also reveal interesting structural differences and unveils an important role of the second extracellular loop in histamine binding.