Functional Characterization of the Epidermal Cholinergic System In Vitro

Kurzen, Hjalmar; Henrich, Carmen; Booken, Dirk; Poenitz, Nina; Gratchev, Alexei; Klemke, Claus‐Detlev; Engstner, Monika; Goerdt, Sergij; Maas‐Szabowski, Nicole

doi:10.1038/sj.jid.5700443

Cited by 41 publications

(37 citation statements)

References 71 publications

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“…2). When given alone, each antagonist produced a severalfold increase of the permeability of monolayers, in keeping with previous studies showing the acantholytic effects of cholinergic antagonists (30,49). Both the PP2A inhibitor OA and the PTP inhibitor pervanadate also considerably increased monolayer permeability (Fig.…”

Section: Roles Of Phosphatases In Cholinergic Receptor-mediated Protesupporting

confidence: 69%

Differential Coupling of M1 Muscarinic and α7 Nicotinic Receptors to Inhibition of Pemphigus Acantholysis

Chernyavsky

Arredondo

Piser

et al. 2008

Journal of Biological Chemistry

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The mechanisms mediating and regulating assembly and disassembly of intercellular junctions is a subject of intensive research. The IgG autoantibodies produced in patients with the immunoblistering skin disease pemphigus vulgaris (PV) can induce keratinocyte (KC) dyshesion (acantholysis) via mechanisms that involve signaling kinases targeting intercellular adhesion molecules, thus providing a useful model to study the physiologic regulation of KC cohesion. Previous studies showed that activation of Src and protein kinase C are the earliest events in the PV IgG-induced intracellular phosphorylation cascades and that cholinergic agonists are effective for treating patients with pemphigus. In this study, we sought to elucidate the molecular mechanisms allowing cholinergic agonists to inhibit PV IgG-induced acantholysis and phosphorylation of KC adhesion molecules. The extent of acantholysis in KC monolayers correlated closely with the degree of PV IgG-induced phosphorylation of p120-and ␤-catenins, with classic isoforms of protein kinase C mediating serine phosphorylation of ␤-catenin and Src-tyrosine phosphorylation of p120-catenin. The M 1 muscarinic agonist pilocarpine blocked phosphorylation of both catenins, which could be abolised by the M 1 antagonist MT7. The ␣7 nicotinic agonist AR-R17779 inhibited phosphorylation of P120-cateinin. The ␣7 antagonist methyllycaconitine abolished the effect of AR-R17779. Okadaic acid abrogated protective effects of agonists on phosphorylation of ␤-catenin, and pervanadate, on that of p120-catenin. Stimulation of KCs with pilocarpine significantly (p < 0.05) elevated both serine/threonine and tyrosine phosphatase activities in KCs. AR-R17779 both stimulated tyrosine phosphatase and decreased PV IgGinduced Src activity. Methyllycaconitine released Src activity in intact KCs and caused acantholysis. Thus, downstream signaling from M 1 abolished PV IgG-dependent catenin phosphorylation due to activation of both serine/threonine and tyrosine phosphatases, whereas ␣7 action involved both activation of tyrosine phosphatase and inhibition of Src. These findings identified novel paradigm of regulation of signaling kinases associated with cholinergic receptors and provided mechanistic explanation of therapeutic activity of cholinomimetics in PV patients.

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Section: Roles Of Phosphatases In Cholinergic Receptor-mediated Protesupporting

confidence: 69%

Differential Coupling of M1 Muscarinic and α7 Nicotinic Receptors to Inhibition of Pemphigus Acantholysis

Chernyavsky

Arredondo

Piser

et al. 2008

Journal of Biological Chemistry

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“…As we could demonstrate previously [15], strychnine alone reduced epidermal thickness to approximately 66% of control (29 m, p<0,001) (Figs. 4d,5).…”

Section: Glycine Induces Epithelial Hyperplasiasupporting

confidence: 57%

“…Immunofluorescence reactions were performed at least 3 times on different specimens according to standard procedures [15]. After blocking unspecific binding sites with 3% BSA in Tris-buffered saline (TBS), a streptavidin blocking solution and a biotin blocking solution (both Dianova, Hamburg, Germany) were applied for 15 minutes each, followed by washing in TBS.…”

Section: Immunofluorescence Stainingmentioning

confidence: 99%

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Glycine Receptors are Present in Human Epidermis

Booken¹,

Henrich-Kellner²,

Klein³

et al. 2008

TODJ

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Abstract:The inhibitory glycine receptor (GlyR) is a member of the nicotinoid receptor superfamily. This heteropentameric Cl -channel is composed of different (1-4) and a -subunit and mediates fast synaptic transmission in the central nervous system. Since glycine, the natural ligand of GlyR has been found to enhance epidermal barrier recovery; we aimed at characterizing GlyR distribution in human skin and their function in skin physiology. We detected differentsubunits and the -GlyR subunit on mRNA and protein level in human skin and cultured keratinocytes and fibroblasts. In cultured human keratinocytes but not in fibroblasts, glycine induced proliferation. Epidermis-equivalents were significantly thicker than control if cultured in the presence of glycine. In human skin, GlyR immunoreactivity (IR) was detected in the upper epidermal layers. In eczema and psoriasis, GlyR IR was reduced in areas with parakeratosis suggesting a role of GlyR in terminal differentiation and epidermal barrier control.Keywords: Epidermal barrier, chloride, strychnine, amino acid, skin. INTROCUCTIONStructurally, glycine (aminoethanoic acid, C 2 H 5 NO 2 ) is the simplest of the 20 standard proteinogenic amino acids, and yet it has a number of important and distinct functions in the body [1]. In addition to its function as part of peptides and proteins, glycine is a major inhibitory neurotransmitter in adult spinal cord and brain stem, where it controls both motor and sensory pathways. After its release from the presynaptic terminals of glycinergic interneurons, glycine exerts multiple functions in the central nervous system, as an inhibitory neurotransmitter through activation of specific, Cl --permeable, ligand-gated ionotropic receptors (GlyR) and as an obligatory co-agonist with glutamate on the activation of N-methyl-D-aspartate (NMDA) receptors [2,3]. The hyperpolarizing action of glycine is selectively antagonized by strychnine, the most potent GlyR antagonist known [4].The GlyR of different species are composed of two glycosylated integral membrane proteins, the 1-4-subunit of 48 kDa and the -subunit of 58 kDa. In addition, in neuronal tissues, there is an associated peripheral membrane protein of 93 kDa named gephyrin [5][6][7]. The primary structures of GlyR -and -subunits deduced by molecular cloning show significant sequence and structural similarity to nicotinic acetylcholine receptor (nAChR), -aminobutyric acid type A (GABA A) receptor, and serotonin type 3 (5HT3) receptor proteins. In situ hybridization showed that the different GlyR subunit genes exhibit unique spatial and temporal expression patterns in spinal cord, brain stem, and some higher brain regions. GlyR 2-subunit transcripts predominate in the embryonic and neonatal brain and spinal cord, but are replaced Recently, several unexpected properties of glycine have been discovered. Like the structurally related nicotinic acetylcholine receptors [8], GlyR were shown to be expressed in peripheral, non-neuronal tissues and cells like macrophages and leukocytes sugge...

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“…28 They offer characteristics that are much closer to the in vivo situation of the skin in comparison to monolayer cell culture systems such as stratification, homeostasis, expression and location of specific differentiation markers. 29,30 It has been shown, that the following parameters of GC-induced skin atrophy can be detected human skin such as the high number of skin appendages and the absence of a papillary dermis in rodents clearly might influence pharmacokinetics. 32 Therefore, absorption of topically administered compounds in rodent skin can exceed human skin by ten times.…”

Section: Determining the Atrophogenic Potential In Pre-clinical In VImentioning

confidence: 99%

Test systems for the determination of glucocorticoid receptor ligand induced skin atrophy

Schoepe

Schäcke

Asadullah

2011

Dermato-Endocrinology

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Functional Characterization of the Epidermal Cholinergic System In Vitro

Cited by 41 publications

References 71 publications

Differential Coupling of M1 Muscarinic and α7 Nicotinic Receptors to Inhibition of Pemphigus Acantholysis

Differential Coupling of M1 Muscarinic and α7 Nicotinic Receptors to Inhibition of Pemphigus Acantholysis

Glycine Receptors are Present in Human Epidermis

Test systems for the determination of glucocorticoid receptor ligand induced skin atrophy

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