Tian, Yuemin and Schreiber, Rainer and Wanitchakool, Podchanart and Kongsuphol, Patthara and Sousa, Marisa and Uliyakina, Inna and Palma, Marta and Faria, Diana and Traynor-Kaplan, Alexis E. and Fragata, Jose I. and Amaral, Margarida D. and Kunzelmann, Karl (2013) Control of TMEM16A by INO-4995 and other inositolphosphates. BRITISH JOURNAL OF PHARMACOLOGY, 168 (1). pp. 253-265. ISSN 0007-1188, 1476-5381
Full text not available from this repository. (Request a copy)Abstract
Background And Purpose Ca2+-dependent Cl- secretion (CaCC) in airways and other tissues is due to activation of the Cl- channel TMEM16A (anoctamin 1). Earlier studies suggested that Ca2+-activated Cl- channels are regulated by membrane lipid inositol phosphates, and that 1-O-octyl-2-O-butyryl-myo-inositol 3,4,5,6-tetrakisphosphate octakis(propionoxymethyl) ester (INO-4995) augments CaCC. Here we examined whether TMEM16A is the target for INO-4995 and if the channel is regulated by inositol phosphates. Experimental Approach The effects of INO-4995 on CaCC were examined in overexpressing HEK293, colonic and primary airway epithelial cells as well as Xenopus oocytes. We used patch clamping, double electrode voltage clamp and Ussing chamber techniques. Key Results We found that INO-4995 directly activates a TMEM16A whole cell conductance of 6.1 +/- 0.9?nS pF1 in overexpressing cells. The tetrakisphosphates Ins(3,4,5,6)P4 or Ins(1,3,4,5)P4 and enzymes controlling levels of InsP4 or PIP2 and PIP3 had no effects on the magnitude or kinetics of TMEM16A currents. In contrast in Xenopus oocytes, human airways and colonic cells, which all express TMEM16A endogenously, Cl- currents were not acutely activated by INO-4995. However incubation with INO-4995 augmented 1.6- to 4-fold TMEM16A-dependent Cl- currents activated by ionomycin or ATP, while intracellular Ca2+ signals were not affected. The potentiating effect of INO-4995 on transient ATP-activated TMEM16A-currents in cystic fibrosis (CF) airways was twice of that observed in non-CF airways. Conclusions And Implications These data indicate that TMEM16A is the target for INO-4995, although the mode of action appears different for overexpressed and endogenous channels. INO-4995 may be useful for the treatment of CF lung disease.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | ACTIVATED CHLORIDE CONDUCTANCE; DEPENDENT PROTEIN-KINASE; PHOSPHATE SIGNALING PATHWAYS; COLONIC EPITHELIAL-CELLS; CYSTIC-FIBROSIS; MYOINOSITOL 3,4,5,6-TETRAKISPHOSPHATE; INOSITOL 3,4,5,6-TETRAKISPHOSPHATE; HUMAN ITPK1; ANNEXIN-IV; LINE T-84; INO-4995; INO4913; anoctamin 1; TMEM16A; inositol phosphates; Ins(3; 4; 5; 6)P4; inositol 3; 4; 5; 6-tetrakisphosphate; Ins(1; 3; 4; 5)P4; inositol 1; 3; 4; 5-tetrakisphosphate; Ca2+-activated Cl- channels; CaCC |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Physiologie Biology, Preclinical Medicine > Institut für Physiologie > Prof. Dr. Karl Kunzelmann |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 30 Apr 2020 05:17 |
| Last Modified: | 30 Apr 2020 05:17 |
| URI: | https://pred.uni-regensburg.de/id/eprint/17549 |
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