Faculty Collaboration Database

Total Matrix Ca²⁺ Modulates Ca²⁺ Efflux via the Ca²⁺/H⁺ Exchanger in Cardiac Mitochondria. Front Physiol 2020;11:510600

Date

10/13/2020

Pubmed ID

33041851

Pubmed Central ID

PMC7526510

DOI

10.3389/fphys.2020.510600

Scopus ID

2-s2.0-85091935444 (requires institutional sign-in at Scopus site)   13 Citations

Abstract

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.

Author List

Natarajan GK, Glait L, Mishra J, Stowe DF, Camara AKS, Kwok WM

Authors

Amadou K. Camara PhD Professor in the Anesthesiology department at Medical College of Wisconsin
Jyotsna Mishra PHD Research Scientist I in the Anesthesiology department at Medical College of Wisconsin
Gayathri K. Natarajan Research Scientist I in the Anesthesiology department at Medical College of Wisconsin
David F. Stowe MD, PhD Professor in the Anesthesiology department at Medical College of Wisconsin