title: alpha 7-type acetylcholine receptor localization and its modulation by nicotine and cholesterol in vascular endothelial cells.
abstract: The neuronal-type α7 nicotinic acetylcholine receptor (α7AChR) is also found in various non-neural tissues, including vascular endothelium, where its peculiar ionotropic properties (high Ca(2+) permeability) and its supervening Ca(2+) -mediated intracellular cascades may play important roles in physiology (angiogenesis) and pathology (inflammation and atherogenesis). Changes in molecular (up-regulation, affinity, and conformational states) and cellular (distribution, association with membranes) properties of the α7AChR related to angiogenesis (wound-repair cell migration) and atherogenesis (alterations in cholesterol content) were studied in living endothelial cells, with the aim of determining whether such changes constitute early markers of inflammatory response. The combination of pharmacological, biochemical, and fluorescence microscopy tools showed that α7AChRs in rat arterial endothelial (RAEC) and human venous endothelial (HUVEC) cells occur at extremely low expression levels (∼50 fmol/mg protein) but undergo agonist-induced up-regulation at relatively high nicotine concentrations (∼300-fold with 50 µM ligand), increasing their cell-surface exposure. When analyzed in terms of cold Triton X-100 solubility and subcellular distribution, α7AChRs occur in the "non-raft" subcellular membrane fractions. Acute cholesterol depletion reduced not only cholesterol levels but also the number of cell-surface α7AChRs. Nicotine exposure markedly stimulated cell migration and accelerated wound repair, which drastically diminished in cells deprived of the sterol. The angiogenic effect of nicotine appears to be synergistic with cholesterol content. Finally, the apparent K(D) of α7AChRs for the open-channel blocker crystal violet was found to be ∼600-fold lower in receptor-enriched membranes obtained from up-regulated HUVEC.
authors: Instituto de Investigaciones Bioquímicas and UNESCO Chair of Biophysics & Molecular Neurobiology, 8000 Bahía Blanca, Argentina.
authors: Victoria B Ayala
authors: VB
authors: Peña
authors: Ida C
authors: IC
authors: Bonini
authors: Silvia S
authors: SS
authors: Antollini
authors: Toshihide
authors: T
authors: Kobayashi
authors: Francisco J
authors: FJ
authors: Barrantes
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D002784:Cholesterol / Q000494:pharmacology*; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D009538:Nicotine / Q000494:pharmacology*; D051381:Rats; D011978:Receptors, Nicotinic / Q000378:metabolism*; D015854:Up-Regulation / Q000187:drug effects; D064569:alpha7 Nicotinic Acetylcholine Receptor
chemical_list: C579167:Chrna7 protein, human; C579165:Chrna7 protein, rat; D011978:Receptors, Nicotinic; D064569:alpha7 Nicotinic Acetylcholine Receptor; D009538:Nicotine; D002784:Cholesterol