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Jean-Quartier, C; Bondarenko, AI; Alam, MR; Trenker, M; Waldeck-Weiermair, M; Malli, R; Graier, WF.
Studying mitochondrial Ca(2+) uptake - a revisit.
Mol Cell Endocrinol. 2012; 353(1-2):114-127
Doi: 10.1016/j.mce.2011.10.033
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- Führende Autor*innen der Med Uni Graz
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Jean-Quartier Claire
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Malli Roland
- Co-Autor*innen der Med Uni Graz
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Alam Muhammad Rizwan
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Bondarenko Oleksandr
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Graier Wolfgang
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Trenker Michael
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Waldeck-Weiermair Markus
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- Abstract:
- Mitochondrial Ca(2+) sequestration is a well-known process that is involved in various physiological and pathological mechanisms. Using isolated suspended mitochondria one unique mitochondrial Ca(2+) uniporter was considered to account ubiquitously for the transfer of Ca(2+) into these organelles. However, by applying alternative techniques for measuring mitochondrial Ca(2+) uptake evidences for molecularly distinct mitochondrial Ca(2+) carriers accumulated recently. Herein we compared different methodical approaches of studying mitochondrial Ca(2+) uptake. Patch clamp technique on mitoplasts from endothelial and HeLa cells revealed the existence of three and two mitoplast Ca(2+) currents (I(CaMito)), respectively. According to their conductance, these channels were named small (s-), intermediate (i-), large (l-) and extra-large (xl-) mitoplast Ca(2+) currents (MCC). i-MCC was found in mitoplasts of both cell types whereas s-MCC and l-MCC or xl-MCC were/was exclusively found in mitoplasts from endothelial cells or HeLa cells. The comparison of mitochondrial Ca(2+) signals, measured either indirectly by sensing extra-mitochondrial Ca(2+) or directly by recording changes of the matrix Ca(2+), showed different Ca(2+) sensitivities of the distinct mitochondrial Ca(2+) uptake routes. Subpopulations of mitochondria with different Ca(2+) uptake capacities in intact endothelial cells could be identified using Rhod-2/AM. In contrast, cells expressing mitochondrial targeted pericam or cameleon (4mtD3cpv) showed homogeneous mitochondrial Ca(2+) signals in response to cell stimulation. The comparison of different experimental approaches and protocols using isolated organelles, permeabilized and intact cells, pointed to cell-type specific and versatile pathways for mitochondrial Ca(2+) uptake. Moreover, this work highlights the necessity of the utilization of multiple technical approaches to study the complexity of mitochondrial Ca(2+) homeostasis.
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Calcium - metabolism
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Endothelial Cells - cytology Endothelial Cells - metabolism
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HeLa Cells -
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Homeostasis - physiology
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Humans -
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Ion Transport - physiology
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Mitochondria - metabolism
- Find related publications in this database (Keywords)
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Mitochondrial Ca2+ measurements
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Mitochondrial Ca2+ uptake
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Mitochondrial Ca2+ uniporter
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Mitoplasts
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Patch clamp
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Pericam