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Title: Agonist-induced down-regulation of type 1 and type 3 inositol 1,4,5-trisphosphate receptors in A7r5 and DDT1 MF-2 smooth muscle cells
Authors: Sipma, Henk ×
Deelman, L
De Smedt, Humbert
Missiaen, Ludwig
Parys, Jan
Vanlingen, Sara
Henning, R H
Casteels, Rik #
Issue Date: Jun-1998
Series Title: Cell calcium. vol:23 issue:1 pages:11-21
Abstract: Prolonged stimulation of rat A7r5 aortic smooth muscle cells with 3 microM vasopressin, or of hamster DDT1 MF-2 smooth muscle cells with 10 microM bradykinin or 100 microM histamine led within 4 h to a 40-50% down-regulation of the type 1 InsP3 receptor (InsP3R-1) and of the type 3 InsP3 receptor (InsP3R-3). InsP3R down-regulation was a cell- and agonist-specific process, since several other agonists acting on PLC-coupled receptors did not change the expression level of the InsP3R isoforms in these cell types and since no agonist-induced down-regulation of InsP3Rs was observed in HeLa cells. Down-regulation of InsP3Rs was prevented by an inhibitor of proteasomal protease activity, N-acetyl-Leu-Leu-norleucinal (ALLN). The Ca2+ channel blocker verapamil (2 microM) also induced InsP3R-1 down-regulation (43%) in A7r5 cells, which was inhibited by ALLN. In A7r5 cells transiently transfected with a cDNA construct, bearing a luciferase coding sequence under control of the rat InsP3R-1 promoter, reduced luciferase activity could be demonstrated upon stimulation of cells with vasopressin or verapamil. Thus, besides enhanced protein degradation, a reduction of InsP3R promoter activity might contribute to the down-regulation of InsP3Rs in A7r5 cells. We next investigated the effect of InsP3R down-regulation on Ca2+ responses in A7r5 cells. A rightward shift in the dose-response curve for InsP3-induced Ca2+ release was observed in permeabilized monolayers of vasopressin-pretreated A7r5 cells (EC50 630 nM and 400 nM for pretreated and non-pretreated cells, respectively). The Ca2+ responses to threshold doses of vasopressin were markedly reduced in intact vasopressin-pretreated cells. We conclude that prolonged agonist-exposure leads to down-regulation of InsP3Rs in A7r5 and DDT, MF-2 smooth muscle cells. The mechanism of down-regulation likely involves proteasomal degradation and reduction of InsP3R promoter activity. Moreover, down-regulation of InsP3Rs resulted in desensitization of Ca2+ release from InsP3 sensitive stores.
ISSN: 0143-4160
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Physiology Section (-)
Laboratory of Molecular and Cellular Signaling
× corresponding author
# (joint) last author

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