Keratinocyte muscarinic acetylcholine receptors: immunolocalization and partial characterization

Details

Serval ID
serval:BIB_5E956CC5CC90
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Keratinocyte muscarinic acetylcholine receptors: immunolocalization and partial characterization
Journal
Journal of Investigative Dermatology
Author(s)
Grando  S. A., Zelickson  B. D., Kist  D. A., Weinshenker  D., Bigliardi  P. L., Wendelschafer-Crabb  G., Kennedy  W. R., Dahl  M. V.
ISSN
0022-202X (Print)
Publication state
Published
Issued date
01/1995
Volume
104
Number
1
Pages
95-100
Notes
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: Jan
Abstract
We have reported previously that human keratinocytes synthesize and secrete acetylcholine and that muscarinic cholinergic drugs have effects on keratinocyte proliferation, adhesion, and migration. This study defines the location of muscarinic acetylcholine receptors in human epidermis and describes some pharmacologic and molecular properties of these receptors. Confocal microscopy employing the anti-muscarinic receptor monoclonal antibody M35 visualized the receptors in the intercellular areas of normal human epidermis. Using immunoelectron microscopy, the receptors appeared to be attached to the keratinocyte plasma membranes. Functional, high-density (Bmax = 8.3 nmol/2 x 10(6) cells) and high-affinity (Kd = 21.5 nM) muscarinic receptors were demonstrated by saturable binding of the reversible radioligand [3H]quinuclidinyl benzilate to the surfaces of freshly isolated epidermal cells at 0 degrees C. Receptor proteins were separated by gel electrophoresis. An apparent isoelectric point of pH 4.3 was determined in immunoblots of sodium-cholate-solubilized receptors separated on isoelectric-focusing gels. Three protein bands, two at approximately 60 kDa and one at 95 kDa, were visualized in immunoblots of membrane-bound or solubilized receptors separated by sodium dodecylsulfate-polyacrylamide gel electrophoresis. The covalent, irreversible ligand [3H]propylbenzilylcholine mustard confirmed these results. Thus, human keratinocytes express a heterogeneous population of muscarinic cholinergic receptors. Because human keratinocytes also express nicotinic cholinergic receptors, endogenously secreted acetylcholine may control different biologic processes in these cells by activating different types of their cholinergic receptors.
Keywords
Antibodies, Monoclonal Blotting, Western Fluorescent Antibody Technique Humans Keratinocytes/*chemistry/ultrastructure Ligands Microscopy, Immunoelectron Molecular Weight Receptors, Muscarinic/*analysis Staining and Labeling
Pubmed
Web of science
Open Access
Yes
Create date
25/01/2008 16:30
Last modification date
20/08/2019 14:16
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