The relationship between the ionized Ca concentration and mitochondrial functions

• Main Authors: L. G. Babich, S. G. Shlykov, A. M. Kushnarova-Vakal, N. I. Kupynyak, V. V. Manko, V. P. Fomin, S. O. Kosterin
• Format: Article
• Language: English
• Published: National Academy of Sciences of Ukraine and Palladin Institute of Biochemistry of the National Academy of Sciences of Ukraine. 2018-04-01
• Series: Ukrainian Biochemical Journal
• Subjects: ionized Са concentration; liver; mitochondria; myometrium; total Са(2+) accumulation
• Online Access: http://ukrbiochemjournal.org/wp-content/uploads/2018/04/Babich_3_18.pdf

The aim of the study was to show the relationships between ionized Ca concentration ([Ca2+]m) in the mitochondria matrix and functional activity of this organelle. [Ca2+]m was determined using the fluorescent probe Fluo-4, AM. Total level of Ca2+ accumulation in mitochondria was monitored using 45Ca2+ as radioactive tracer. It was shown that incubation of myometrium mitochondria with 3 mM Mg2+ resulted in the low level of [Ca2+]m. Subsequent addition of 100 µM Ca2+ resulted in 8 times increase of [Ca2+]m but in low level of total calcium accumulation. Normalized fluorescence of Ca2+-sensitive probe Fluo-4 in response to the Ca2+ addition was higher than 2.5. At the same time, [Ca2+]m was considerably higher in the medium containing­ 3 mМ АТР and 3 mМ Mg2+. Subsequent addition of 100 µM Ca2+ to the incubation medium resulted in only 2.4 times increase of [Ca2+]m but considerably higher level of total calcium accumulation was observed. Normali­zed fluorescence of Fluo-4 in response to the Ca2+ addition was lower than 1.3. In liver mitochondria higher rate of oxygen consumption was detected in the presence of an oxidative substrate succinate than of pyruvate or α-ketoglutarate. At the presence of an oxidative substrate succinate normalized fluorescence of Fluo-4 in liver mitochondria in response to the Ca2+ addition was lower than 1.3. It was concluded that low level of [Ca2+]m was correlated with low functional activity of this organelle and, vise versa, high level of [Ca2+]m was correlated with high functional activity. It was suggested that normalized fluorescence changes in response to the Са2+ addition could be used as a test of the mitochondrial functional activity: lower normalized fluorescence values − higher functional activity.