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Microscopic analysis of Orai-mediated store-operated calcium entry in cells with experimentally altered levels of amyloid precursor protein
Familial Alzheimer's disease (FAD)-causing mutations in presenilins were shown to alter intracellular calcium dynamics, including store-operated calcium entry (SOCE). However, the involvement of FAD-linked amyloid precursor protein (APP) in SOCE remains controversial. Here, we used gain-of-func...
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Published in: | Biochemical and biophysical research communications 2016-09, Vol.478 (3), p.1087-1092 |
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Main Authors: | , , |
Format: | Article |
Language: | English |
Subjects: | |
Citations: | Items that this one cites Items that cite this one |
Online Access: | Get full text |
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Summary: | Familial Alzheimer's disease (FAD)-causing mutations in presenilins were shown to alter intracellular calcium dynamics, including store-operated calcium entry (SOCE). However, the involvement of FAD-linked amyloid precursor protein (APP) in SOCE remains controversial. Here, we used gain-of-function and loss-of-function approaches to shed light on this issue. We found that Jurkat cells, which exhibit prominent SOCE mediated by Orai channels, maintain low APP levels. The ectopic expression of APP, either with wildtype sequence or FAD-causing Swedish mutation, had no effect on SOCE induced by calcium store depletion with cyclopiazonic acid (CPA). The overproduction of C99 fragments, mimicking amyloidogenic processing of APP, also had no effect. Moreover, there was no alteration in the CPA-evoked SOCE upon APP knockdown in HeLa cells, which natively express 100-fold more APP than Jurkat cells. Consistently, we found no evidence for APP-dependent changes in the mRNA or protein levels of main SOCE components. Altogether, these results suggest that APP does not modulate Orai-dependent SOCE following quantitative calcium store depletion.
•The APP gene is weakly expressed in Jurkat cells, 100-fold less than in HeLa cells.•APP expression does not affect SOCE or STIM1/Orai1 protein levels in Jurkat cells.•APP knockdown does not affect SOCE or STIM1/ORAI1 mRNA levels in HeLa cells. |
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ISSN: | 0006-291X 1090-2104 |
DOI: | 10.1016/j.bbrc.2016.08.072 |