Muscarinic receptors on intact human fibroblasts. Absence of receptor activity in adult skin cells.

Riper, D A Van; Absher, Marlene; Lenox, R H

doi:10.1172/jci112047

Cited by 15 publications

(5 citation statements)

References 11 publications

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“…In 1984, Nadi et al demonstrated that intact cultured adult human skin fibroblasts possessed muscarinic receptors that could be labelled by [3H]QNB (Nadi et al, 1984). In contrast, Van Riper et al (1985) and Kelsoe et al (1986) showed that these cells have almost negligible levels of [3H]QNB binding (Kelsoe et al, 1986;Van Riper et al, 1985). However, Kelsoe et al (1986) Nadi et al, whereas the Kd values were almost identical.…”

Section: Discussionmentioning

confidence: 99%

Characterization of muscarinic acetylcholine receptors in cultured adult skin fibroblasts: Effects of the swedish Alzheimer's disease APP 670/671 mutation on binding levels

Vestling

Cowburn

Venizelos

et al. 1995

J Neural Transm Gen Sect

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We have characterised the muscarinic receptor subtypes found in human skin fibroblasts and compared binding levels in cell lines from members of the Alzheimer's disease family with the Swedish amyloid precursor protein (APP) 670/671 mutation. Binding studies with [3H] quinuclidinyl benzilate ([3H]QNB) and the M2/M4 selective antagonist [3H] (+/-)-5,11-dihydro-11-([(2-[(di-propylamino)methyl]-1- piperidinyl]ethyl)amino]carbonyl)-6H-pyrido(2,3-b)(1,4)benzodiazepine-6- one ([3H]AF-DX 384) revealed the presence of a single population of muscarinic receptors on lysed fibroblast membranes. [3H]QNB binding was displaced by a number of selective muscarinic ligands with a rank order of potency: atropine > himbacine > methoctramine > (+/-)-p-fluoro-hexahydro-sila-difenidol hydrochloride > pirenzepine > muscarinic-toxin-3. APP 670/671 mutation carrying cell lines showed 25-35% lower levels of muscarinic receptors labelled with [3H]QNB, [3H]N-methyl scopolamine and [3H]AF-DX 384, compared to controls. This difference was not statistically significant due to large individual variation. It is concluded that muscarinic receptors on adult skin fibroblasts are predominantly of the M2 subtype. Since these cells do not possess M1 and M3 receptor subtypes, they are unlikely to provide a good model for studying muscarinic receptor regulation of APP processing.

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Section: Discussionmentioning

confidence: 99%

Characterization of muscarinic acetylcholine receptors in cultured adult skin fibroblasts: Effects of the swedish Alzheimer's disease APP 670/671 mutation on binding levels

Vestling

Cowburn

Venizelos

et al. 1995

J Neural Transm Gen Sect

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“…In agreement with this, Vestling et al (1995) have identified M 1 and M 3 as the causes of agonist‐stimulated DNA synthesis in fibroblasts. However, using the reverse transcription‐polymerase chain reaction and Northern blot technique the m 2 , m 4 and m 5 mAChR subtypes were identified human skin fibroblast surface (Buchli et al, 1999), but only negligible activity of these receptors was documented (Van Riper et al, 1985).…”

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confidence: 99%

Dynamic cell shapes and contacts in the developing Drosophila retina are regulated by the Ig cell adhesion protein hibris

Grillo-Hill

Wolff

2009

Developmental Dynamics

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Cell shapes and contacts are dynamically regulated during organogenesis to enable contacts with relevant neighboring cells at appropriate times. During Drosophila larval eye development, an apical contact is established between one pair of non-neuronal cones cells, precluding contact between the opposing pair. Concurrent with changes in cell shape, these contacts reverse in early pupal life. The reversal in cone cell contacts occurs in a posterior to anterior gradient across the eye, following the developmental gradient established in the larval eye imaginal disc. Hibris (Hbs), an Immunoglobulin cell adhesion molecule homologous to vertebrate Nephrin, is required for cone cell morphogenesis. In hbs null mutants, a majority of cone cells fail to both establish wild-type contacts and achieve mature cone cell shapes. hbs acts cell autonomously in the cone cells to drive these changes. The work presented here indicates hbs contributes to the remodeling of cell contacts and cell shapes throughout development. Developmental Dynamics 238: 2223-2234, 2009.

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“…Vestling et al [1995] also demonstrated high levels of saturable binding of [ 3 H]QNB and the M2/M4 selective radiolabel [ 3 H]AF-DX 384 to lysed HSF membrane preparations. By contrast, Van Riper et al [1985], Lin and Richelson [1986], and Kelsoe et al [1986] reported that fibroblasts have only negligible amounts of muscarinic ligand binding sites. Difficulties with studying fibroblast mAChRs have been acknowledged by a number of investigators [e.g., Lennox et al, 1985].…”

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confidence: 99%

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

et al. 1999

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Previous studies have demonstrated that muscarinic acetylcholine receptors (mAChRs) are expressed by human skin fibroblasts (HSF). We have identified the molecular subtypes of these receptors by reverse transcription-polymerase chain reaction (RT-PCR), using m1-m5 subtype-specific primers. These experiments showed that only mRNAs for m2, m4, and m5 mAChR subtypes are present in HSF. The RT-PCR products were characterized by restriction analysis and Southern blotting. Northern blot analysis showed the presence of m2 and m4 mAChR RNA. Rabbit antibodies were raised using a synthetic peptide as immunogen corresponding to the C-terminus of the m2 protein and were used to visualize fibroblast mAChRs. Cell membranes of HSF in cell culture and specimens of normal human skin had a unique staining pattern specific for anti-m2 antibody, as well as for antibodies against m4 and m5. In Western blots of fibroblast proteins, the antibodies visualized the m2 receptor at 65 kDa, m4 at 70 kDa, and m5 at 95 kDa. The function of fibroblast mAChRs was examined by measuring muscarinic effects on intracellular free Ca2+ concentration ([Ca2+]i). Muscarine increased transiently [Ca2+]i in cultured HSF. This effect could be abolished by the muscarinic antagonist atropine. Thus, the results of this study showed that HSF express m2, m4, and m5 mAChR subtypes, and that fibroblast mAChRs are coupled to the regulation of [Ca2+]i.

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Muscarinic receptors on intact human fibroblasts. Absence of receptor activity in adult skin cells.

Abstract: The intact human fibroblast has been used in clinical and basic research studies of receptor-mediated control of cell function, however there is little information about the relationship between muscarinic receptor density and the regulation of cyclic AMP (cAMP)

Cited by 15 publications

References 11 publications

Characterization of muscarinic acetylcholine receptors in cultured adult skin fibroblasts: Effects of the swedish Alzheimer's disease APP 670/671 mutation on binding levels

Characterization of muscarinic acetylcholine receptors in cultured adult skin fibroblasts: Effects of the swedish Alzheimer's disease APP 670/671 mutation on binding levels

Dynamic cell shapes and contacts in the developing Drosophila retina are regulated by the Ig cell adhesion protein hibris

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

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