Human skin fibroblasts express m2, m4, and m5 subtypes of muscarinic acetylcholine receptors

Buchli, Rico; Ndoye, Assane; Rodriguez, Jennifer; Zia, Shaheen; Webber, Robert J.; Grando, Sergei A.

doi:10.1002/(sici)1097-4644(19990801)74:2<264::aid-jcb11>3.0.co;2-z

Cited by 57 publications

(32 citation statements)

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“…ACh, Nic, and cholinergic compounds elicit biologic effects on DF through binding to two different classes of cholinergic receptors, the nicotinic and the muscarinic types of ACh receptors. In addition to nAChRs characterized in this study, DF also express m2, m4, and m5 muscarinic ACh receptor subtypes that are coupled to the regulation of Ca 2ϩ metabolism in these cells (Buchli et al, 1999). In addition to DF, the two classes of ACh receptors coexist on the cell membranes of several types of non-neuronal cells and are functionally interrelated (Bering et al, 1987;Evinger et al, 1994;Falugi et al, 1993;Richman and Arnason, 1979;Stewart and Forrester, 1978;Young and Laing, 1991).…”

Section: Arredondo Et Almentioning

confidence: 62%

Central Role of Fibroblast α3 Nicotinic Acetylcholine Receptor in Mediating Cutaneous Effects of Nicotine

Arredondo

Hall

Ndoye

et al. 2003

Laboratory Investigation

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SUMMARY:Smoking is associated with aberrant cutaneous tissue remodeling, such as precocious skin aging and impaired wound healing. The mechanism is not fully understood. Dermal fibroblasts (DF) are the primary cellular component of the dermis and may provide a target for pathobiologic effects of tobacco products. The purpose of this study was to characterize a mechanism of nicotine (Nic) effects on the growth and tissue remodeling function of DF. We hypothesized that the effects of Nic on DF result from its binding to specific nicotinic acetylcholine receptors (nAChRs) expressed by these cells and that downstream signaling from the receptors alters normal cell functioning, leading to changes in skin homeostasis. Using RT-PCR and Western blotting, we found that a 24-hour exposure of human DF to 10 M Nic causes a 1.9-to 28-fold increase of the mRNA and protein levels of the cell cycle regulators p21, cyclin D1, Ki-67, and PCNA and a 1.7-to 2-fold increase of the apoptosis regulators Bcl-2 and caspase 3. Nic exposure also up-regulated expression of the dermal matrix proteins collagen type I␣1 and elastin as well as matrix metalloproteinase-1. Mecamylamine (Mec), the specific antagonist of nAChRs, abolished Nic-induced alterations, indicating that they resulted from a pharmacologic stimulation of nAChRs expressed by DF. To establish the relevance of these findings to a specific nicotinergic pathway, we studied human DF transfected with anti-␣3 antisense oligonucleotides and murine DF from ␣3 nAChR knockout mice. In both cases, lack of ␣3 was associated with alterations in fibroblast growth and function that were opposite to those observed in DF treated with Nic, suggesting that the nicotinic effects on DF were mostly mediated by ␣3 nAChR. In addition to ␣3, the nAChR subunits detected in human DF were ␣5, ␣7, ␤2, and ␤4. The exposure of DF to Nic altered the relative amounts of each of these subunits, leading to reciprocal changes in [ 3 H]epibatidine-binding kinetics. Thus, some of the pathobiologic effects of tobacco products on extracellular matrix turnover in the skin may stem from Nic-induced alterations in the physiologic control of the unfolding of the genetically determined program of growth and the tissue remodeling function of DF as well as alterations in the structure and function of fibroblast nAChRs. (Lab Invest 2003, 83:207-225).

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Section: Arredondo Et Almentioning

confidence: 62%

Central Role of Fibroblast α3 Nicotinic Acetylcholine Receptor in Mediating Cutaneous Effects of Nicotine

Arredondo

Hall

Ndoye

et al. 2003

Laboratory Investigation

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100

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“…Hassall et al (1993) could not detect the M 5 mRNA in intrinsic cardiac ganglia using in situ hybridization. Reports on the presence of M 5 mRNA in rat arteries (Phillips et al, 1997), human brain microvascular endothelial cells (Elhusseiny et al, 1999), and human skin fibroblasts (Buchli et al, 1999) suggest that cardiac endothelial cells and fibroblasts should be considered among potential sources of the M 5 mRNA we detected in the heart.…”

Section: Discussionmentioning

confidence: 91%

Quantitation of mRNAs for M₁to M₅Subtypes of Muscarinic Receptors in Rat Heart and Brain Cortex

Krejčı́

Tuček

2002

Mol Pharmacol

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It has been generally accepted that, of the five subtypes of muscarinic receptors (M 1 -M 5 ), only the M 2 subtype is expressed in mammalian heart. This notion has recently been challenged by a series of reports indicating that mRNAs for some or all non-M 2 subtypes are also present in mammalian heart, in parallel with the M 2 mRNA. However, the quantities of relevant mRNAs reported to be present in the heart are not known, which makes it difficult to evaluate their likely significance. We measured the concentrations of the five muscarinic mRNAs by competitive reverse transcription-polymerase chain reaction and discovered that the M 2 mRNA represents more than 90% of total muscarinic mRNAs in rat atria and in either ventricle. The concentrations of total muscarinic mRNAs and of the M 2 mRNA were more than twice as high in the atria than in the ventricles. mRNAs for all non-M 2 muscarinic receptor subtypes were also detected but represented less than 1% (M 1 and M 4 ), less than 3% (M 3 ), and less than 5% (M 5 ) of total muscarinic RNAs in the atria and ventricles. The findings support the concept of the prevalent role of the M 2 muscarinic receptors in the cholinergic control of the heart. When the same method of quantitation was applied to rat cerebral cortex, mRNAs for individual subtypes were found to represent 36% (M 1 ), 21% (M 2 ), 25% (M 3 ), 11% (M 4 ), and 7% (M 5 ) of total muscarinic mRNAs.

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“…Total RNA and protein in SCC and the normal lung tissues were prepared with TRI-reagent (Molecular Research Center, Inc.; ref. 26). The probes and primers used in real-time PCRs were as described previously (11) or are listed in Supplementary Table S1.…”

Section: Methodsmentioning

confidence: 99%

Activated Cholinergic Signaling Provides a Target in Squamous Cell Lung Carcinoma

et al. 2008

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The binding of exogenous nicotine to nicotinic acetylcholine (ACh) receptors (nAChR) and the binding of endogenous ACh to both nAChR and muscarinic ACh receptors (mAChR) stimulate growth of both small cell and non-small cell lung carcinomas. Understanding how cholinergic signaling is upregulated in lung cancer may suggest new therapeutic approaches. Analysis of 28 squamous cell lung carcinomas (SCC) showed increased levels of A5 and B3 nAChR mRNA and increased levels of ACh associated with increased levels of choline acetyltransferase mRNA and decreased cholinesterase mRNAs. Lynx1, an allosteric inhibitor of nAChR activity, was also decreased in SCC. Thus, cholinergic signaling is broadly increased in SCC caused by increased levels of receptors, increased levels of ligands, and decreased levels of receptor inhibitors. Partially explaining the cholinergic up-regulation seen in SCC, incubation of the H520 SCC cell line with nicotine increased levels of ACh secretion, increased expression of nAChR, and, as measured by electrophysiologic recording, increased activity of the expressed nAChR. Consistent with these effects, nicotine stimulated proliferation of H520 cells. One approach to blocking proliferative effects of nicotine and ACh on growth of lung cancers may be through M3 mAChR antagonists, which can limit the activation of mitogenactivated protein kinase that is caused by both nicotinic and muscarinic signaling. This was tested with the M3-selective muscarinic antagonist darifenacin. Darifenacin blocked nicotine-stimulated H520 growth in vitro and also blocked H520 growth in nude mice in vivo. Thus, cholinergic signaling is broadly up-regulated in SCC and blocking cholinergic signaling can limit basal and nicotine-stimulated growth of SCC.

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Human skin fibroblasts express m2, m4, and m5 subtypes of muscarinic acetylcholine receptors

Cited by 57 publications

References 45 publications

Central Role of Fibroblast α3 Nicotinic Acetylcholine Receptor in Mediating Cutaneous Effects of Nicotine

Central Role of Fibroblast α3 Nicotinic Acetylcholine Receptor in Mediating Cutaneous Effects of Nicotine

Quantitation of mRNAs for M₁to M₅Subtypes of Muscarinic Receptors in Rat Heart and Brain Cortex

Activated Cholinergic Signaling Provides a Target in Squamous Cell Lung Carcinoma

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Human skin fibroblasts express m2, m4, and m5 subtypes of muscarinic acetylcholine receptors

Cited by 57 publications

References 45 publications

Central Role of Fibroblast α3 Nicotinic Acetylcholine Receptor in Mediating Cutaneous Effects of Nicotine

Central Role of Fibroblast α3 Nicotinic Acetylcholine Receptor in Mediating Cutaneous Effects of Nicotine

Quantitation of mRNAs for M1to M5Subtypes of Muscarinic Receptors in Rat Heart and Brain Cortex

Activated Cholinergic Signaling Provides a Target in Squamous Cell Lung Carcinoma

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Quantitation of mRNAs for M₁to M₅Subtypes of Muscarinic Receptors in Rat Heart and Brain Cortex