All muscarinic acetylcholine receptors (M1-M5) are expressed in murine brain microvascular endothelium

Abstract

Clinical and experimental studies indicate that muscarinic acetylcholine receptors are potential pharmacological targets for the treatment of neurological diseases. Although these receptors have been described in human, bovine and rat cerebral microvascular tissue, a subtype functional characterization in mouse brain endothelium is lacking. Here, we show that all muscarinic acetylcholine receptors (M₁-M₅) are expressed in mouse brain microvascular endothelial cells. The mRNA expression of M₂, M₃, and M₅ correlates with their respective protein abundance, but a mismatch exists for M₁ and M₄ mRNA versus protein levels. Acetylcholine activates calcium transients in brain endothelium via muscarinic, but not nicotinic, receptors. Moreover, although M₁ and M₃ are the most abundant receptors, only a small fraction of M₁ is present in the plasma membrane and functions in ACh-induced Ca²⁺ signaling. Bioinformatic analyses performed on eukaryotic muscarinic receptors demonstrate a high degree of conservation of the orthosteric binding site and a great variability of the allosteric site. In line with previous studies, this result indicates muscarinic acetylcholine receptors as potential pharmacological targets in future translational studies. We argue that research on drug development should especially focus on the allosteric binding sites of the M₁ and M₃ receptors.