| 2018 |
Chrm1 and Chrm3 double knockout in mice chronically diminishes REM sleep to an almost undetectable level, establishing that these two Gq-type muscarinic acetylcholine receptors are essential for REM sleep generation. |
Triple-target CRISPR knockout of acetylcholine receptor genes in mice with EEG/sleep phenotype readout |
Cell reports |
High |
30157420
|
| 2015 |
Autocrine CHRM3 activation by acetylcholine in prostate epithelial cells promotes cancer cell proliferation, migration, and castration resistance via calmodulin/calmodulin-dependent protein kinase kinase (CaM/CaMKK)-mediated phosphorylation of Akt. |
CHRM3 overexpression, shRNA knockdown, carbachol activation, darifenacin pharmacological blockade, Western blotting for phospho-Akt; in vitro and xenograft models |
Clinical cancer research |
Medium |
26071486
|
| 2020 |
Choline activates M3AChR (CHRM3) on vascular smooth muscle cells (VSMCs), promoting Nrf2 nuclear translocation and induction of HO-1 and NQO-1, thereby attenuating Ang II-induced ROS production, VSMC phenotypic switching, proliferation, and migration; downregulation of M3AChR or Nrf2 abolishes these protective effects. |
siRNA knockdown of M3AChR and Nrf2 in cultured VSMCs, Nrf2 nuclear translocation assay, Western blotting for HO-1/NQO-1/NOX, abdominal aortic constriction rat model |
Arteriosclerosis, thrombosis, and vascular biology |
Medium |
32938216
|
| 2021 |
Pancreas-specific activation of a chemogenetic mutant CHRM3 (responsive to CNO but not acetylcholine) in acinar cells is sufficient to induce acute pancreatitis in mice, and CHRM3 antagonist ameliorates cerulein-induced pancreatitis in wild-type mice; CHRM3 activation elicits intracellular calcium release and amylase secretion in acinar cells. |
Conditional transgenic expression of DREADD mutant M3R in pancreatic acinar cells, CNO-induced calcium/amylase assays, histopathology, M3R antagonist rescue |
JCI insight |
High |
34314386
|
| 2019 |
In L-arginine-induced severe acute pancreatitis, CHRM3 activation promotes acinar cell necrosis by suppressing miR-31-5p, which leads to upregulation of RIP3 via the MAPK-p38 pathway; CRISPR/Cas9-mediated Chrm3 knockout reduces pathological lesions and serum amylase activity. |
CRISPR/Cas9 Chrm3 knockout mice, PLV-EGFP-Chrm3 overexpression in acinar cells, miR-31-5p expression analysis, RIP3 Western blotting, MAPK-p38 pathway assessment |
Pancreas |
Medium |
33122522
|
| 2019 |
Chrm3 is expressed on pancreatic acinar cell membranes and stabilizes caspase-8 expression to suppress necrosis (but not apoptosis) in L-arginine-induced severe acute pancreatitis; Chrm3 reduction decreased pathological lesions and serum amylase. |
CRISPR/Cas9-mediated Chrm3 knockout, immunolocalization of Chrm3 in islets and acinar cell membranes, caspase-8 Western blotting, cell death assays |
Journal of cellular biochemistry |
Medium |
31692054
|
| 2019 |
CHRM3 is the direct molecular target for miR-15b-5p; miR-15b-5p binds CHRM3 mRNA directly and suppresses CHRM3-mediated YAP activation and castration-resistant prostate cancer cell growth; androgen deprivation suppresses miR-15b-5p and increases CHRM3 expression, establishing a miR-15b-5p/CHRM3/YAP signaling axis. |
Small RNA sequencing, in silico target prediction, luciferase reporter assay for direct miR-15b-5p binding to CHRM3, YAP activation assay, CRPC cell growth assay |
FEBS letters |
Medium |
36754848
|
| 2013 |
In bovine tracheal smooth muscle plasma membranes, cGMP/PKG-II-dependent phosphorylation of M3AChR (CHRM3) modulates its ligand-binding properties: cGMP increases [3H]QNB binding activity stimulated by ATP, causes disappearance of high-affinity agonist binding sites, and reveals an additional high-affinity antagonist (4-DAMP) binding site; 4-DAMP-mustard alkylation of CHRM3 blocks the cGMP effect, establishing CHRM3 as the main target of cGMP-dependent phosphorylation in airway smooth muscle. |
Radioligand binding assay with [3H]QNB, 4-DAMP-mustard alkylation, PKG-II inhibitor/activator pharmacology, Western blotting for PKG-II, phosphorylation assay |
Molecular membrane biology |
Medium |
24175711
|
| 2022 |
CHRM3 hypermethylation in human umbilical vein following IVF-ET leads to reduced CHRM3 expression, decreased PKCβ and CaV1.2 protein levels, reduced MLC20 phosphorylation, and attenuated acetylcholine-induced vasoconstriction, establishing a CHRM3-CaV1.2/PKC-MLC20 phosphorylation signaling pathway in umbilical vascular smooth muscle. |
Vascular tone measurement with selective receptor antagonists, Western blotting, quantitative real-time PCR, methylation analysis of CHRM3 promoter in human umbilical vein tissue |
Biology of reproduction |
Medium |
34935917
|
| 2022 |
The VP1 protein of porcine circovirus-like virus P1 physically interacts with CHRM3, and through this interaction activates the pancreatic secretion pathway (including Gq, PLC-β2, PRKCA, Rab3D, RhoA, Rac1, and amyA downstream components), promoting secretion of digestive enzymes. |
Co-IP, GST pull-down, confocal fluorescence microscopy; transfection of P1 plasmid and virus infection in PK-15 and 3D4 cells; RT-PCR and Western blotting for pathway components |
Veterinary microbiology |
Medium |
35810687
|
| 2022 |
In spinal cord ventral horn neurons, pilocarpine activates M3-AChR (blocked by selective antagonists 4-DAMP and J104129) and increases glycine-induced currents through the IP3/intracellular Ca2+/PKC signaling pathway; chelating intracellular Ca2+ (BAPTA) or blocking IP3 receptors (heparin, Xe-C) or PKC (Bis-IV) abolished the effect, and PKC activator (PMA) mimicked it. |
Patch-clamp electrophysiology of isolated spinal cord neurons, selective M3-AChR antagonists, IP3R antagonists, PKC inhibitor/activator pharmacology, Ca2+-free extracellular solution and BAPTA chelation |
Neuroscience letters |
Medium |
35598692
|
| 2021 |
Choline activates M3AChR to promote HSF1 nuclear translocation and Notch1 intracellular domain (NICD) expression in cardiomyocytes, attenuating Ang II-induced mitochondrial ROS production and apoptosis; knockdown of M3AChR or Notch1 blocks choline-mediated cardioprotection, establishing M3AChR→Notch1/HSF1 as the upstream signaling mechanism. |
siRNA knockdown of M3AChR and Notch1 in neonatal rat cardiomyocytes, HSF1 nuclear translocation assay, mtROS measurement, TUNEL assay, in vivo rat Ang II infusion model |
Oxidative medicine and cellular longevity |
Medium |
34504645
|
| 2025 |
The RING E3 ubiquitin ligase PJA2 physically interacts with CHRM3, ubiquitinates it, and targets it for proteasomal degradation, thereby suppressing downstream TGFβ-pSMAD3 signaling and gastric tumor cell progression; catalytically dead ΔRING PJA2 mutant cannot suppress CHRM3 signaling or PDO growth. |
Mass spectrometry and Co-IP to identify PJA2-CHRM3 interaction; ubiquitination assay; degradation assay; ΔRING mutant rescue; patient-derived organoid (PDO) viability; xenograft metastasis model |
British journal of cancer |
High |
40858831
|
| 2025 |
CHRM3 overexpression activates PI3K/AKT and MAPK signaling pathways and increases proliferation, migration, and invasion of lung cancer cells; CHRM3 knockdown reduces these pathways and phenotypes. |
Stable CHRM3 overexpression and knockdown via lentiviral infection in lung cancer lines, Western blotting for PI3K/AKT and MAPK pathway proteins, proliferation/migration/invasion assays, xenograft tumor model |
Frontiers in oncology |
Medium |
40718823
|
| 2025 |
In human umbilical vein from GDM pregnancies, CHRM3 is upregulated due to promoter hypomethylation, leading to enhanced ACh-induced vasoconstriction; pretreatment with P-F-HHSiD (selective CHRM3 antagonist) abolishes the GDM-control difference in ACh-induced constriction, establishing CHRM3 as the primary mediator of the enhanced vascular response. |
Organ bath vascular tone measurements with selective muscarinic receptor subtype antagonists, methylation assay of CHRM3 promoter, Western blotting and RT-PCR for CHRM3 and CACNA1C in human umbilical vein tissue |
Scientific reports |
Medium |
40624113
|
| 2025 |
In human memory T helper cells, CHRM3 (M3R) is required for mAChR-stimulated increases in IFN-γ and IL-17A (pro-inflammatory cytokines) and NF-κB p65 activation; CHRM3 knockout (CRISPR-Cas9) or M3R-selective blockade (J104129) prevented increases in IFN-γ and IL-17A but continued to suppress IL-4. |
CRISPR-Cas9 CHRM3 knockout, M3R-selective antagonist J104129, RT-qPCR for CHRM1-5, Western blot for M3R and phospho-NF-κB p65, ELISA and intracellular cytokine staining for IFN-γ/IL-17A/IL-4 |
bioRxivpreprint |
Medium |
|
| 2025 |
In macaque prefrontal and anterior cingulate cortices, CHRM3 is the most enriched mAChR gene in neurons but is largely retained in the nucleus, in contrast to CHRM1 which traffics to the cytoplasm; CHRM3 and CHRM1 show strong co-expression and functional overlap in modulating excitatory:inhibitory synaptic ratio and spine morphology. |
Single-nucleus RNA sequencing, mRNA-protein histology, in vitro functional experiments measuring E:I synaptic ratio and spine morphology in ACC vs LPFC neurons |
bioRxivpreprint |
Medium |
|
| 2024 |
Pharmacological blockade of M1 and M3 muscarinic acetylcholine receptors (including CHRM3) abolishes activity-regulated DMG (diffuse midline glioma) proliferation in cholinergic neuron-glioma co-culture and in vivo, demonstrating that CHRM3 mediates cholinergic neuron-driven glioma proliferation. |
hiPSC-derived cholinergic neuron co-culture with DMG cells, pharmacological blockade of M1/M3 receptors, in vivo optogenetic stimulation of cholinergic midbrain nuclei, single-cell RNA sequencing for CHRM3 expression in DMG |
bioRxivpreprint |
Medium |
|