Dynamic Regulation of the Mitochondrial Proton Gradient during Cytosolic Calcium Elevations

Poburko, Damon; Santo‐Domingo, Jaime; Demaurex, Nicolas

doi:10.1074/jbc.m110.159962

Cited by 284 publications

(294 citation statements)

References 71 publications

Supporting

Mentioning

271

Contrasting

Unclassified

Order By: Relevance

“…The LETM1-encoding plasmid (35) was provided by Dr. Luca Scorrano (University of Geneva). The mitochondrial pH sensor mitoSypHer was described previously (52).…”

Section: Methodsmentioning

confidence: 99%

NCLX Protein, but Not LETM1, Mediates Mitochondrial Ca2+ Extrusion, Thereby Limiting Ca2+-induced NAD(P)H Production and Modulating Matrix Redox State

Marchi

Santo‐Domingo

Castelbou

et al. 2014

Journal of Biological Chemistry

Self Cite

103

View full text Add to dashboard Cite

Background: Whether mitochondrial Ca 2ϩ extrusion is mediated by NCLX (mitochondrial sodium/calcium exchanger) or LETM1 (leucine zipper-EF-hand-containing transmembrane protein 1) and controls matrix redox state is unknown. Results: NCLX, but not LETM1, increases Ca 2ϩ extrusion, limits NAD(P)H production, and promotes matrix oxidation. Conclusion: NCLX controls the duration of matrix Ca 2ϩ elevations and their impact on redox signaling. Significance: NCLX is a potential target for the treatment of redox-dependent diseases.

show abstract

“…The LETM1-encoding plasmid (35) was provided by Dr. Luca Scorrano (University of Geneva). The mitochondrial pH sensor mitoSypHer was described previously (52).…”

Section: Methodsmentioning

confidence: 99%

NCLX Protein, but Not LETM1, Mediates Mitochondrial Ca2+ Extrusion, Thereby Limiting Ca2+-induced NAD(P)H Production and Modulating Matrix Redox State

Marchi

Santo‐Domingo

Castelbou

et al. 2014

Journal of Biological Chemistry

Self Cite

103

View full text Add to dashboard Cite

show abstract

“…pH in the mitochondrial matrix is estimated at~8.0 (40), but direct visualization of spatiotemporal pH dynamics using a genetically encoded mitochondrial pH indicator revealed two unique features of mitochondrial matrix pH: (1) an acidification during cytosolic Ca 2þ elevation by the activation of Ca 2þ / H þ transporters and (2) a spontaneous and transient alkalization, named pH flash (49,50). These mitochondrial pH changes have the potential to cause artifactual fluctuations in the fluorescent intensity of pH-sensitive GECIs.…”

Section: Mitochondriamentioning

confidence: 99%

Genetically Encoded Fluorescent Indicators for Organellar Calcium Imaging

Suzuki

Kanemaru

Iino

2016

Biophysical Journal

115

View full text Add to dashboard Cite

Optical Ca 2þ indicators are powerful tools for investigating intracellular Ca 2þ signals in living cells. Although a variety of Ca 2þ indicators have been developed, deciphering the physiological functions and spatiotemporal dynamics of Ca 2þ in intracellular organelles remains challenging. Genetically encoded Ca 2þ indicators (GECIs) using fluorescent proteins are promising tools for organellar Ca 2þ imaging, and much effort has been devoted to their development. In this review, we first discuss the key points of organellar Ca 2þ imaging and summarize the requirements for optimal organellar Ca 2þ indicators. Then, we highlight some of the recent advances in the engineering of fluorescent GECIs targeted to specific organelles. Finally, we discuss the limitations of currently available GECIs and the requirements for advancing the research on intraorganellar Ca 2þ signaling.

show abstract

“…Acidification of the cellular environment leads to protonation, thus mimicking reduction of the probe; alkalization on the other hand mimics oxidation (Belousov et al 2006). SypHer, a H 2 O 2 -insensitive HyPer variant (C199S) is a pH sensor and can be used as a control (Poburko et al 2011). Another family of fluorescent protein based redox-sensitive probes are the roGFPs, which were created by introducing oxidizable cysteine residues on the outside of the ß-barrel 8 structure of GFP near the location of the chromophore.…”

Section: Sensorsmentioning

confidence: 99%

Integrated High-Content Quantification of Intracellular ROS Levels and Mitochondrial Morphofunction

Sieprath

Corne

Willems

et al. 2016

Advances in Anatomy, Embryology and Cell Biology

View full text Add to dashboard Cite

Oxidative stress arises from an imbalance between the production of reactive oxygen species (ROS) and their removal by cellular antioxidant systems. Especially under pathological conditions, mitochondria constitute a relevant source of cellular ROS. These organelles harbor the electron transport chain, bringing electrons in close vicinity to molecular oxygen. Although a full understanding is still lacking, intracellular ROS generation and mitochondrial function are also linked to changes in mitochondrial morphology. To study the intricate relationships between the different factors that govern cellular redox balance in living cells, we have developed a high-content microscopy-based strategy for simultaneous quantification of intracellular ROS levels and mitochondrial morphofunction. Here, we summarize the principles of intracellular ROS generation and removal, and we explain the major considerations for performing quantitative microscopy analyses of ROS and mitochondrial morphofunction in living cells. Next, we describe our workflow and finally, we illustrate that a multiparametric readout enables the unambiguous classification of chemically perturbed cells as well as laminopathy patient cells. Principles of intracellular ROS generation and removalReactive oxygen species (ROS) are small, short-lived derivatives of molecular oxygen (O 2 ) of radical and non-radical nature (Halliwell and Gutteridge 2007 • ), nitrate radical (NO 3 • ) and many other nitrogen derivatives. ROS were originally described as molecular constituents of the defense system of phagocytic cells, but it has become clear that besides their damaging properties, they also function as signaling molecules and mediate a variety of other cellular responses including cell proliferation, differentiation, gene expression and migration (Lambeth 2004;Bartz and Piantadosi 2010). Intracellular ROS metabolismROS can be generated at various sites in the cell (Fig. 1A). This can be either deliberately, e.g. by NADPH oxidases (NOX), or as a byproduct, e.g. during normal cellular respiration in mitochondria (Babior 1999;Turrens 2003;Murphy 2009). The NOX family of NADPH oxidases (NOX1, NOX2, NOX3, NOX4, NOX5, DUOX1 and DUOX2) are proteins that transport electrons (e -) from NADPH across biological membranes (plasma or endomembranes) (Bedard and Krause 2007;Dupre-Crochet et al. 2013). The activation mechanisms and tissue distribution of the isoforms differ but they all use O 2 as e --acceptor, producing O 2•-. Through ROS generation, they play a role in many cellular processes including host defense, regulation of gene expression and cell differentiation (Bedard and Krause 2007). Despite their sometimes significant contribution to the global ROS pools, NOX are not the predominant source of intracellular ROS. Mitochondria are considered the major culprit, in particular under pathological conditions. Mitochondrial ROS are generated as a byproduct of the oxidative phosphorylation (OXPHOS, cf. below). Irrespective of its source, ROS production generally sta...

show abstract

Dynamic Regulation of the Mitochondrial Proton Gradient during Cytosolic Calcium Elevations

Cited by 284 publications

References 71 publications

NCLX Protein, but Not LETM1, Mediates Mitochondrial Ca2+ Extrusion, Thereby Limiting Ca2+-induced NAD(P)H Production and Modulating Matrix Redox State

NCLX Protein, but Not LETM1, Mediates Mitochondrial Ca2+ Extrusion, Thereby Limiting Ca2+-induced NAD(P)H Production and Modulating Matrix Redox State

Genetically Encoded Fluorescent Indicators for Organellar Calcium Imaging

Integrated High-Content Quantification of Intracellular ROS Levels and Mitochondrial Morphofunction

Contact Info

Product

Resources

About