Ousingsawat, Jiraporn and Cabrita, Ines and Wanitchakool, Podchanart and Sirianant, Lalida and Krautwald, Stefan and Linkermann, Andreas and Schreiber, Rainer and Kunzelmann, Karl (2017) Ca2+ signals, cell membrane disintegration, and activation of TMEM16F during necroptosis. CELLULAR AND MOLECULAR LIFE SCIENCES, 74 (1). pp. 173-181. ISSN 1420-682X, 1420-9071
Full text not available from this repository. (Request a copy)Abstract
Activated receptor-interacting protein kinase 3 (RIPK3) and mixed lineage kinase domain like (MLKL) are essential components of the necroptotic pathway. Phosphorylated MLKL (pMLKL) is thought to induce membrane leakage, leading to cell swelling and disintegration of the cell membrane. However, the molecular identity of the necroptotic membrane pore remains unclear, and the role of pMLKL for membrane permeabilization is currently disputed. We observed earlier that the phospholipid scramblase and ion channel TMEM16F/anoctamin 6 cause large membrane currents, cell swelling, and cell death when activated by a strong increase in intracellular Ca2+. We, therefore, asked whether TMEM16F is also central to necroptotic cell death and other cellular events during necroptosis. Necroptosis was induced by TNF alpha, smac mimetic, and Z-VAD (TSZ) in NIH3T3 fibroblasts and the four additional cell lines HT29, 16HBE, H441, and L929. Time-dependent changes in intracellular Ca2+, cell morphology, and membrane currents were recorded. TSZ induced a small and only transient oscillatory rise in intracellular Ca2+, which was paralleled by the activation of outwardly rectifying Cl- currents, which were typical for TMEM16F/ANO6. Ca2+ oscillations were due to Ca2+ release from endoplasmic reticulum, and were independent of extracellular Ca2+. The initial TSZ-induced cell swelling was followed by cell shrinkage. Using typical channel blockers and siRNA-knockdown, the Cl- currents were shown to be due to the activation of ANO6. However, the knockdown of ANO6 or inhibitors of ANO6 did not inhibit necroptotic cell death. The present data demonstrate the activation of ANO6 during necroptosis, which, however, is not essential for cell death.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | DIGITAL HOLOGRAPHIC MICROSCOPY; CA2+-ACTIVATED CL-CHANNELS; REGULATED ANION CHANNEL; ESSENTIAL COMPONENT; CHLORIDE CHANNELS; ANOCTAMIN 6; DEATH; PROTEIN; NECROSIS; VRAC; Cell death; Necroptosis; Apoptosis; TMEM16F; Anoctamin 6; Chloride channel |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Biology, Preclinical Medicine > Institut für Physiologie > Prof. Dr. Karl Kunzelmann |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 14 Dec 2018 12:58 |
| Last Modified: | 14 Feb 2019 13:46 |
| URI: | https://pred.uni-regensburg.de/id/eprint/134 |
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