Muscarinic Receptor Trafficking

Reiner, Cindy; Nathanson, Neil M.

doi:10.1007/978-3-642-23274-9_4

Cited by 10 publications

(10 citation statements)

References 105 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…All of the subtypes are expressed in the central nervous system but, among them, M1 is the major subtype in cortical and hippocampal neurons (Levey et al, 1995;Levey et al, 1991). Trafficking of mAChRs is regulated in a subtypeand/or cell-type-specific manner (Nathanson, 2008;Reiner and Nathanson, 2012). Interestingly, recent immunohistochemical and pharmacological studies have shown stable intracellular localization of M1-mAChR, especially in neuronal cells (Anisuzzaman et al, 2013;Wang et al, 1994;Yamasaki et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

Intracellular localization of M1 muscarinic acetylcholine receptor through clathrin-dependent constitutive internalization via a C-terminal tryptophan-based motif

Uwada¹,

Yoshiki²,

Masuoka³

et al. 2014

Journal of Cell Science

View full text Add to dashboard Cite

The M1 muscarinic acetylcholine receptor (M1-mAChR, encoded by CHRM1) is a G-protein-coupled membrane receptor that is activated by extracellular cholinergic stimuli. Recent investigations have revealed the intracellular localization of M1-mAChR. In this study, we observed constitutive internalization of M1-mAChR in mouse neuroblastoma N1E-115 cells without agonist stimulation. Constitutive internalization depended on dynamin, clathrin and the adaptor protein-2 (AP-2) complex. A WxxI motif in the M1-mAChR C-terminus is essential for its constitutive internalization, given that replacement of W 442 or I 445 with alanine residues abolished constitutive internalization. This WxxI motif resembles YxxW, which is the canonical binding motif for the m2 subunit of the AP-2 complex. The M1-mAChR C-terminal WxxI motif interacted with AP-2 m2. W442A and I445A mutants of the M1-mAChR C-terminal sequence lost AP-2-m2-binding activity, whereas the W442Y mutant bound more effectively than wild type. Consistent with these results, W442A and I445A M1-mAChR mutants selectively localized to the cell surface. By contrast, the W442Y receptor mutant was found only at intracellular sites. Our data indicate that the cellular distribution of M1-mAChR is governed by the C-terminal tryptophanbased motif, which mediates constitutive internalization.

show abstract

Section: Introductionmentioning

confidence: 99%

Intracellular localization of M1 muscarinic acetylcholine receptor through clathrin-dependent constitutive internalization via a C-terminal tryptophan-based motif

Uwada¹,

Yoshiki²,

Masuoka³

et al. 2014

Journal of Cell Science

View full text Add to dashboard Cite

show abstract

“…An accumulation of evidence has revealed the molecular details and regulation of clathrin-endocytic GPCRs, while the molecular mechanisms of nonclathrin-endocytic GPCRs are less clear 3 35 . In this study, we investigated agonist-stimulated internalization and intracellular transport of two structurally and functionally homologous mAChRs, M2 and M4.…”

Section: Discussionmentioning

confidence: 99%

“…The endocytic pathway strongly controls the activity of G-protein-coupled receptors (GPCRs) 1 2 3 . Many GPCRs are subject to the classical clathrin-endocytic pathway, which involves dynamin, β-arrestin, and clathrin-coated pits 2 4 .…”

mentioning

confidence: 99%

“…After internalization, molecular sorting of GPCRs through divergent membrane pathways can result in highly different functional consequences 11 . Elucidating the mechanisms that determine the specificity of GPCR membrane traffic in the endocytic pathway will offer new insights into the regulation of GPCR functions 3 .…”

mentioning

confidence: 99%

“…After agonist induction, the M2 mAChR is internalized via a clathrin-independent endocytosis (CIE) pathway, suggested to be regulated by Arf6 9 and then targeted to lysosomes for degradation 20 . In contrast, the M4 mAChR is internalized through the clathrin-dependent endocytosis (CDE) pathway 3 and then recycled back to the plasma membrane, a process that involves Myosin Vb and Rab11 21 . The M2 and M4 mAChRs show 58% sequence homology; however, alignment of the amino acid sequences within the i3 loop regions of each receptor shows that their composition is divergent, with a shared identity of only 24%.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Unraveling a molecular determinant for clathrin-independent internalization of the M2 muscarinic acetylcholine receptor

Wan

Zhang

Tian

et al. 2015

Sci Rep

View full text Add to dashboard Cite

Endocytosis and postendocytic sorting of G-protein-coupled receptors (GPCRs) is important for the regulation of both their cell surface density and signaling profile. Unlike the mechanisms of clathrin-dependent endocytosis (CDE), the mechanisms underlying the control of GPCR signaling by clathrin-independent endocytosis (CIE) remain largely unknown. Among the muscarinic acetylcholine receptors (mAChRs), the M4 mAChR undergoes CDE and recycling, whereas the M2 mAChR is internalized through CIE and targeted to lysosomes. Here we investigated the endocytosis and postendocytic trafficking of M2 mAChR based on a comparative analysis of the third cytoplasmic domain in M2 and M4 mAChRs. For the first time, we identified that the sequence 374KKKPPPS380 servers as a sorting signal for the clathrin-independent internalization of M2 mAChR. Switching 374KKKPPPS380 to the i3 loop of the M4 mAChR shifted the receptor into lysosomes through the CIE pathway; and therefore away from CDE and recycling. We also found another previously unidentified sequence that guides CDE of the M2 mAChR, 361VARKIVKMTKQPA373, which is normally masked in the presence of the downstream sequence 374KKKPPPS380. Taken together, our data indicate that endocytosis and postendocytic sorting of GPCRs that undergo CIE could be sequence-dependent.

show abstract

The deletion of M₄ muscarinic receptors increases motor activity in females in the dark phase

et al. 2018

View full text Add to dashboard Cite

ObjectivesM4 muscarinic receptors (MR) presumably play a role in motor coordination. Previous studies have shown different results depending on genetic background and number of backcrosses. However, no attention has been given to biorhythms.Material and MethodsWe therefore analyzed biorhythms under a light/dark cycle obtained telemetrically in intact animals (activity, body temperature) in M4 KO mice growth on the C57Bl6 background using ChronosFit software. Studying pure effects of gene knockout in daily rhythms is especially important knowledge for pharmacological/behavioral studies in which drugs are usually tested in the morning.ResultsWe show that M4 KO mice motor activity does not differ substantially from wild‐type mice during light period while in the dark phase (mice active part of the day), the M4 KO mice reveal biorhythm changes in many parameters. Moreover, these differences are sex‐dependent and are evident in females only. Mesor, night–day difference, and night value were doubled or tripled when comparing female KO versus male KO. Our in vitro autoradiography demonstrates that M4 MR proportion represents 24% in the motor cortex (MOCx), 30% in the somatosensory cortex, 50% in the striatum, 69% in the thalamus, and 48% in the intergeniculate leaflet (IGL). The M4 MR densities were negligible in the subparaventricular zone, the posterior hypothalamic area, and in the suprachiasmatic nuclei.ConclusionsWe conclude that cholinergic signaling at M4 MR in brain structures such as striatum, MOCx, and probably with the important participation of IGL significantly control motor activity biorhythm. Animal activity differs in the light and dark phases, which should be taken into consideration when interpreting the results.

show abstract

Muscarinic Receptor Trafficking

Cited by 10 publications

References 105 publications

Intracellular localization of M1 muscarinic acetylcholine receptor through clathrin-dependent constitutive internalization via a C-terminal tryptophan-based motif

Intracellular localization of M1 muscarinic acetylcholine receptor through clathrin-dependent constitutive internalization via a C-terminal tryptophan-based motif

Unraveling a molecular determinant for clathrin-independent internalization of the M2 muscarinic acetylcholine receptor

The deletion of M₄ muscarinic receptors increases motor activity in females in the dark phase

Contact Info

Product

Resources

About

Muscarinic Receptor Trafficking

Cited by 10 publications

References 105 publications

Intracellular localization of M1 muscarinic acetylcholine receptor through clathrin-dependent constitutive internalization via a C-terminal tryptophan-based motif

Intracellular localization of M1 muscarinic acetylcholine receptor through clathrin-dependent constitutive internalization via a C-terminal tryptophan-based motif

Unraveling a molecular determinant for clathrin-independent internalization of the M2 muscarinic acetylcholine receptor

The deletion of M4 muscarinic receptors increases motor activity in females in the dark phase

Contact Info

Product

Resources

About

The deletion of M₄ muscarinic receptors increases motor activity in females in the dark phase