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Distinct properties of Ca²⁺ efflux from brain, heart and liver mitochondria: The effects of Na⁺, Li⁺ and the mitochondrial Na⁺/Ca²⁺ exchange inhibitor CGP37157.

ABSTRACT: Mitochondrial Ca²⁺ transport is essential for regulating cell bioenergetics, Ca²⁺ signaling and cell death. Mitochondria accumulate Ca²⁺ via the mitochondrial Ca²⁺ uniporter (MCU), whereas Ca²⁺ is extruded by the mitochondrial Na⁺/Ca²⁺ (mtNCX) and H⁺/Ca²⁺ exchangers. The balance between these processes is essential for preventing toxic mitochondrial Ca²⁺ overload. Recent work demonstrated that MCU activity varies significantly among tissues, likely reflecting tissue-specific Ca²⁺ signaling and energy needs. It is less clear whether this diversity in MCU activity is matched by tissue-specific diversity in mitochondrial Ca²⁺ extrusion. Here we compared properties of mitochondrial Ca²⁺ extrusion in three tissues with prominent mitochondria function: brain, heart and liver. At the transcript level, expression of the Na⁺/Ca²⁺/Li⁺ exchanger (NCLX), which has been proposed to mediate mtNCX transport, was significantly greater in liver than in brain or heart. At the functional level, Na⁺ robustly activated Ca²⁺ efflux from brain and heart mitochondria, but not from liver mitochondria. The mtNCX inhibitor CGP37157 blocked Ca²⁺ efflux from brain and heart mitochondria but had no effect in liver mitochondria. Replacement of Na⁺ with Li⁺ to test the involvement of NCLX, resulted in a slowing of mitochondrial Ca²⁺ efflux by ∼70 %. Collectively, our findings suggest that mtNCX is responsible for Ca²⁺ extrusion from the mitochondria of the brain and heart, but plays only a small, if any, role in mitochondria of the liver. They also reveal that Li⁺ is significantly less effective than Na⁺ in driving mitochondrial Ca²⁺ efflux.

SUBMITTER: Rysted JE

PROVIDER: S-EPMC8187304 | biostudies-literature | 2021 Jun

REPOSITORIES: biostudies-literature

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Distinct properties of Ca2+ efflux from brain, heart and liver mitochondria: The effects of Na+, Li+ and the mitochondrial Na+/Ca2+ exchange inhibitor CGP37157.

Rysted Jacob E JE Lin Zhihong Z Walters Grant C GC Rauckhorst Adam J AJ Noterman Maria M Liu Guanghao G Taylor Eric B EB Strack Stefan S Usachev Yuriy M YM

Cell calcium 20210222

Mitochondrial Ca2+ transport is essential for regulating cell bioenergetics, Ca2+ signaling and cell death. Mitochondria accumulate Ca2+ via the mitochondrial Ca2+ uniporter (MCU), whereas Ca2+ is extruded by the mitochondrial Na+/Ca2+ (mtNCX) and H+/Ca2+ exchangers. The balance between these processes is essential for preventing toxic mitochondrial Ca2+ overload. Recent work demonstrate ...[more]

PMID: 33684833

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Project description:The cardiac sodium (Na+)/calcium (Ca2+) exchanger (NCX1) is an electrogenic membrane transporter that regulates Ca2+ homeostasis in cardiomyocytes, serving mainly to extrude Ca2+ during diastole. The direction of Ca2+ transport reverses at membrane potentials near that of the action potential plateau, generating an influx of Ca2+ into the cell. Therefore, there has been great interest in the possible roles of NCX1 in cardiac Ca2+-induced Ca2+ release (CICR). Interest has been reinvigorated by a recent super-resolution optical imaging study suggesting that ~18% of NCX1 co-localize with ryanodine receptor (RyR2) clusters, and ~30% of additional NCX1 are localized to within ~120nm of the nearest RyR2. NCX1 may therefore occupy a privileged position in which to modulate CICR. To examine this question, we have developed a mechanistic biophysically-detailed model of NCX1 that describes both NCX1 transport kinetics and Ca2+-dependent allosteric regulation. This NCX1 model was incorporated into a previously developed super-resolution model of the Ca2+ spark as well as a computational model of the cardiac ventricular myocyte that includes a detailed description of CICR with stochastic gating of L-type Ca2+ channels and RyR2s, and that accounts for local Ca2+ gradients near the dyad via inclusion of a peri-dyadic (PD) compartment. Both models predict that increasing the fraction of NCX1 in the dyad and PD decreases spark frequency, fidelity, and diastolic Ca2+ levels. Spark amplitude and duration are less sensitive to NCX1 spatial redistribution. On the other hand, NCX1 plays an important role in promoting Ca2+ entry into the dyad, and hence contributing to the trigger for RyR2 release at depolarized membrane potentials and in the presence of elevated local Na+ concentration. Whole-cell simulation of NCX1 tail currents are consistent with the finding that a relatively high fraction of NCX1 (~45%) resides in the dyadic and PD spaces, with a dyad-to-PD ratio of roughly 1:2. Allosteric Ca2+ activation of NCX1 helps to "functionally localize" exchanger activity to the dyad and PD by reducing exchanger activity in the cytosol thereby protecting the cell from excessive loss of Ca2+ during diastole.

Project description:Mitochondrial Ca2+ handling is accomplished by balancing Ca2+ uptake, primarily via the Ru360-sensitive mitochondrial calcium uniporter (MCU), Ca2+ buffering in the matrix and Ca2+ efflux mainly via Ca2+ ion exchangers, such as the Na+/Ca2+ exchanger (NCLX) and the Ca2+/H+ exchanger (CHE). The mechanism of CHE in cardiac mitochondria is not well-understood and its contribution to matrix Ca2+ regulation is thought to be negligible, despite higher expression of the putative CHE protein, LETM1, compared to hepatic mitochondria. In this study, Ca2+ efflux via the CHE was investigated in isolated rat cardiac mitochondria and permeabilized H9c2 cells. Mitochondria were exposed to (a) increasing matrix Ca2+ load via repetitive application of a finite CaCl2 bolus to the external medium and (b) change in the pH gradient across the inner mitochondrial membrane (IMM). Ca2+ efflux at different matrix Ca2+ loads was revealed by inhibiting Ca2+ uptake or reuptake with Ru360 after increasing number of CaCl2 boluses. In Na+-free experimental buffer and with Ca2+ uptake inhibited, the rate of Ca2+ efflux and steady-state free matrix Ca2+ [mCa2+]ss increased as the number of administered CaCl2 boluses increased. ADP and cyclosporine A (CsA), which are known to increase Ca2+ buffering while maintaining a constant [mCa2+]ss, decreased the rate of Ca2+ efflux via the CHE, with a significantly greater decrease in the presence of ADP. ADP also increased Ca2+ buffering rate and decreased [mCa2+]ss. A change in the pH of the external medium to a more acidic value from 7.15 to 6.8∼6.9 caused a twofold increase in the Ca2+ efflux rate, while an alkaline change in pH from 7.15 to 7.4∼7.5 did not change the Ca2+ efflux rate. In addition, CHE activation was associated with membrane depolarization. Targeted transient knockdown of LETM1 in permeabilized H9c2 cells modulated Ca2+ efflux. The results indicate that Ca2+ efflux via the CHE in cardiac mitochondria is modulated by acidic buffer pH and by total matrix Ca2+. A mechanism is proposed whereby activation of CHE is sensitive to changes in both the matrix Ca2+ buffering system and the matrix free Ca2+ concentration.

Dataset Information

Distinct properties of Ca²⁺ efflux from brain, heart and liver mitochondria: The effects of Na⁺, Li⁺ and the mitochondrial Na⁺/Ca²⁺ exchange inhibitor CGP37157.

Publications

Distinct properties of Ca<sup>2+</sup> efflux from brain, heart and liver mitochondria: The effects of Na<sup>+</sup>, Li<sup>+</sup> and the mitochondrial Na<sup>+</sup>/Ca<sup>2+</sup> exchange inhibitor CGP37157.

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