Abstract
Endoplasmic reticulum (ER) Ca2+ depletion activates the unfolded protein response (UPR), inhibits bulk autophagy
and eventually induces cell death in mammalian cells. However, the extent and duration of ER Ca2+
depletion required is unknown. We instigated a detailed study in two different cell lines, using sarco/endoplasmic
reticulum Ca2+-ATPase (SERCA) inhibitors to gradually reduce ER Ca2+ levels in a controlled
manner. Remarkably, UPR induction (as assessed by expression analyses of UPR-regulated proteins) and autophagy
inhibition (as assessed by analyses of effects on starvation-induced bulk autophagy) required substantially
higher drug concentrations than those needed to strongly decrease total ER Ca2+ levels. In fact, even
when ER Ca2+ levels were so low that we could hardly detect any release of Ca2+ upon challenge with ER Ca2+
purging agents, UPR was not induced, and starvation-induced bulk autophagy was still fully supported.
Moreover, although we observed reduced cell proliferation at this very low level of ER Ca2+, cells could tolerate
prolonged periods (days) without succumbing to cell death. Addition of increasing concentrations of extracellular
EGTA also gradually depleted the ER of Ca2+, and, as with the SERCA inhibitors, EGTA-induced activation
of UPR and cell death required higher EGTA concentrations than those needed to strongly reduce ER Ca2+
levels. We conclude that ER Ca2+ depletion-induced effects on UPR, autophagy and cell death require either an
extreme general depletion of ER Ca2+ levels, or Ca2+ depletion in areas of the ER that have a higher resistance to
Ca2+ drainage than the bulk of the ER.
and eventually induces cell death in mammalian cells. However, the extent and duration of ER Ca2+
depletion required is unknown. We instigated a detailed study in two different cell lines, using sarco/endoplasmic
reticulum Ca2+-ATPase (SERCA) inhibitors to gradually reduce ER Ca2+ levels in a controlled
manner. Remarkably, UPR induction (as assessed by expression analyses of UPR-regulated proteins) and autophagy
inhibition (as assessed by analyses of effects on starvation-induced bulk autophagy) required substantially
higher drug concentrations than those needed to strongly decrease total ER Ca2+ levels. In fact, even
when ER Ca2+ levels were so low that we could hardly detect any release of Ca2+ upon challenge with ER Ca2+
purging agents, UPR was not induced, and starvation-induced bulk autophagy was still fully supported.
Moreover, although we observed reduced cell proliferation at this very low level of ER Ca2+, cells could tolerate
prolonged periods (days) without succumbing to cell death. Addition of increasing concentrations of extracellular
EGTA also gradually depleted the ER of Ca2+, and, as with the SERCA inhibitors, EGTA-induced activation
of UPR and cell death required higher EGTA concentrations than those needed to strongly reduce ER Ca2+
levels. We conclude that ER Ca2+ depletion-induced effects on UPR, autophagy and cell death require either an
extreme general depletion of ER Ca2+ levels, or Ca2+ depletion in areas of the ER that have a higher resistance to
Ca2+ drainage than the bulk of the ER.
| Originalsprog | Engelsk |
|---|---|
| Artikelnummer | https://doi.org/10.1016/j.ceca.2018.09.005 |
| Tidsskrift | Cell Calcium |
| Vol/bind | 76 |
| Sider (fra-til) | 48-61 |
| Antal sider | 14 |
| ISSN | 0143-4160 |
| DOI | |
| Status | Udgivet - dec. 2018 |
Emneord
- Det Sundhedsvidenskabelige Fakultet
- Endoplasmic reticulum
- Calcium
- Unfolded Protein Response
- Thapsigargin
- Cell Death
- Autophagy