scholarly journals Exploring the In Vivo Role of the Mitochondrial Calcium Uniporter in Brown Fat Bioenergetics

Cell Reports ◽  
2019 ◽  
Vol 27 (5) ◽  
pp. 1364-1375.e5 ◽  
Author(s):  
Daniel Flicker ◽  
Yasemin Sancak ◽  
Eran Mick ◽  
Olga Goldberger ◽  
Vamsi K. Mootha
Keyword(s):  
Brown Fat ◽  
Mitochondrial Calcium ◽  
Mitochondrial Calcium Uniporter ◽  
Calcium Uniporter

scholarly journals The physiological role of mitochondrial calcium revealed by mice lacking the mitochondrial calcium uniporter

2013 ◽  
Vol 15 (12) ◽  
pp. 1464-1472 ◽  
Author(s):  
Xin Pan ◽  
Jie Liu ◽  
Tiffany Nguyen ◽  
Chengyu Liu ◽  
Junhui Sun ◽  
...  
Keyword(s):  
Physiological Role ◽  
Mitochondrial Calcium ◽  
Mitochondrial Calcium Uniporter ◽  
Calcium Uniporter

scholarly journals A novel protoapigenone analog RY10-4 induces apoptosis of breast cancer cells by regulating mitochondrial Ca2+ influx through the mitochondrial calcium uniporter

2020 ◽  
Author(s):  
Pingping Xue ◽  
Qian Chen ◽  
Xiuhua Ren ◽  
Yimin Yang ◽  
Xiaofan Yang ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cancer Cells ◽  
Breast Cancer Cells ◽  
Flavonoid Compound ◽  
Mitochondrial Calcium ◽  
Mitochondrial Apoptosis ◽  
Apoptosis Pathway ◽  
Mitochondrial Calcium Uniporter ◽  
Calcium Uniporter

Abstract Background Protoapigenone, as a flavonoid compound with a specific nonaromatic B-ring, exhibits extraordinary antitumor activities against a broad spectrum of human cancer cells. Here we developed a novel protoapigenone analog RY10-4, which induces the apoptosis of various tumor cells, especially for breast cancer cells, but the underlying mechanism involved in the apoptotic process remains unclear. Methods MTT assay, colony-formation assay and flow cytometry were applied to evaluate the proliferation and apoptosis of breast cancer cells. Cytoplasmic calcium ([Ca2+]c) and mitochondrial calcium ([Ca2+]m) of the breast cancer cells were measured by the Fluo-2 and Rhod-2 probes, respectively. The mitochondrial reactive oxygen species (mtROS), membrane potential (ΔΨm) and permeability transition pore (mPTP) were analyzed by MitoSOX, JC-1 probes and Calcein/AM, respectively. Furthermore, Western bolt assay was adopted for the exploration of the mitochondrial apoptosis pathway. Besides, the xenograft assay was performed to investigate the role of RY10-4 in breast cancer cells in vivo. Results Obviously, RY10-4 could effectively suppress the proliferation and induce the apoptosis of breast cancer cells. Furthermore, the [Ca2+]c and [Ca2+]m of MDA-MB-231 cells were up-regulated after the treatment of RY10-4, resulting in the mtROS accumulation, ΔΨm depolarization and mPTP opening. And finally, the mitochondrial apoptosis was activated by the release of cytochrome C. Interestingly, the inhibition of mitochondrial calcium uniporter (MCU) with Ru360 attenuated the overload of [Ca2+]m and blocked the apoptosis of MDA-MB-231 cells induced by RY10-4, which was also consistent with the in vivo results. Conclusions From the results we concluded that RY10-4 could induce apoptosis of breast cancer cells by elevating [Ca2+]m through MCU. Our work contributed previously unknown insights into the mechanisms involving in the clinical efficacy of RY10-4 on breast cancer cells, which also advanced calcium homeostasis as a potential target for chemotherapeutic drugs.

scholarly journals Actin enhances ER-mitochondrial calcium transfer and IMM constriction during mitochondrial division

2017 ◽  
Author(s):  
Rajarshi Chakrabarti ◽  
Wei-Ke Ji ◽  
Radu V. Stan ◽  
Jaime de Juan Sanz ◽  
Timothy A. Ryan ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Actin Polymerization ◽  
Mitochondrial Calcium ◽  
Mitochondrial Membranes ◽  
Independent Manner ◽  
Mitochondrial Division ◽  
Mitochondrial Calcium Uniporter ◽  
Calcium Uniporter ◽  
Transport Chain

SummaryMitochondrial division requires division of both the inner and outer mitochondrial membranes (IMM and OMM, respectively). Interaction with endoplasmic reticulum (ER) promotes OMM division by recruitment of the dynamin Drp1, but effects on IMM division are not well characterized. We previously showed that actin polymerization through the ER-bound formin INF2 stimulates Drp1 recruitment in mammalian cells. Here, we show that INF2-mediated actin polymerization stimulates a second mitochondrial response independent of Drp1: a rise in mitochondrial matrix calcium through the mitochondrial calcium uniporter. ER stores supply the increased mitochondrial calcium, and the role of actin is to increase ER-mitochondria contact. Myosin IIA is also required for this mitochondrial calcium increase. Elevated mitochondrial calcium in turn activates IMM constriction in a Drp1-independent manner. IMM constriction requires electron transport chain activity. IMM division precedes OMM division. These results demonstrate that actin polymerization independently stimulates the dynamics of both membranes during mitochondrial division: IMM through increased matrix calcium, and OMM through Drp1 recruitment.

scholarly journals MICU1 and MICU2 Play an Essential Role in Mitochondrial Ca2+ Uptake, Growth, and Infectivity of the Human Pathogen Trypanosoma cruzi

mBio ◽  
2019 ◽  
Vol 10 (3) ◽  
Author(s):  
Mayara S. Bertolini ◽  
Miguel A. Chiurillo ◽  
Noelia Lander ◽  
Anibal E. Vercesi ◽  
Roberto Docampo
Keyword(s):  
Trypanosoma Cruzi ◽  
Cell Lines ◽  
Citrate Synthase ◽  
Host Cells ◽  
Mitochondrial Calcium ◽  
Content Type ◽  
Mitochondrial Calcium Uniporter ◽  
Calcium Uniporter ◽  
Eukaryotic Supergroup

ABSTRACT The mitochondrial Ca2+ uptake in trypanosomatids, which belong to the eukaryotic supergroup Excavata, shares biochemical characteristics with that of animals, which, together with fungi, belong to the supergroup Opisthokonta. However, the composition of the mitochondrial calcium uniporter (MCU) complex in trypanosomatids is quite peculiar, suggesting lineage-specific adaptations. In this work, we used Trypanosoma cruzi to study the role of orthologs for mitochondrial calcium uptake 1 (MICU1) and MICU2 in mitochondrial Ca2+ uptake. T. cruzi MICU1 (TcMICU1) and TcMICU2 have mitochondrial targeting signals, two canonical EF-hand calcium-binding domains, and localize to the mitochondria. Using the CRISPR/Cas9 system (i.e., clustered regularly interspaced short palindromic repeats with Cas9), we generated TcMICU1 and TcMICU2 knockout (-KO) cell lines. Ablation of either TcMICU1 or TcMICU2 showed a significantly reduced mitochondrial Ca2+ uptake in permeabilized epimastigotes without dissipation of the mitochondrial membrane potential or effects on the AMP/ATP ratio or citrate synthase activity. However, none of these proteins had a gatekeeper function at low cytosolic Ca2+ concentrations ([Ca2+]cyt), as occurs with their mammalian orthologs. TcMICU1-KO and TcMICU2-KO epimastigotes had a lower growth rate and impaired oxidative metabolism, while infective trypomastigotes have a reduced capacity to invade host cells and to replicate within them as amastigotes. The findings of this work, which is the first to study the role of MICU1 and MICU2 in organisms evolutionarily distant from animals, suggest that, although these components were probably present in the last eukaryotic common ancestor (LECA), they developed different roles during evolution of different eukaryotic supergroups. The work also provides new insights into the adaptations of trypanosomatids to their particular life styles. IMPORTANCE Trypanosoma cruzi is the etiologic agent of Chagas disease and belongs to the early-branching eukaryotic supergroup Excavata. Its mitochondrial calcium uniporter (MCU) subunit shares similarity with the animal ortholog that was important to discover its encoding gene. In animal cells, the MICU1 and MICU2 proteins act as Ca2+ sensors and gatekeepers of the MCU, preventing Ca2+ uptake under resting conditions and favoring it at high cytosolic Ca2+ concentrations ([Ca2+]cyt). Using the CRISPR/Cas9 technique, we generated TcMICU1 and TcMICU2 knockout cell lines and showed that MICU1 and -2 do not act as gatekeepers at low [Ca2+]cyt but are essential for normal growth, host cell invasion, and intracellular replication, revealing lineage-specific adaptations.

scholarly journals The mitochondrial calcium uniporter is crucial for the generation of fast cortical network rhythms

2019 ◽  
Vol 40 (11) ◽  
pp. 2225-2239 ◽  
Author(s):  
Carlos Bas-Orth ◽  
Justus Schneider ◽  
Andrea Lewen ◽  
Jamie McQueen ◽  
Kerstin Hasenpusch-Theil ◽  
...  
Keyword(s):  
Energy Homeostasis ◽  
Pyramidal Cells ◽  
Gamma Oscillations ◽  
Spike Timing ◽  
Mitochondrial Calcium ◽  
Attention And Memory ◽  
Mitochondrial Calcium Uniporter ◽  
Sharp Waves ◽  
Calcium Uniporter

The role of the mitochondrial calcium uniporter (MCU) gene ( Mcu) in cellular energy homeostasis and generation of electrical brain rhythms is widely unknown. We investigated this issue in mice and rats using Mcu-knockout and -knockdown strategies in vivo and in situ and determined the effects of these genetic manipulations on hippocampal gamma oscillations (30–70 Hz) and sharp wave-ripples. These physiological network states require precise neurotransmission between pyramidal cells and inhibitory interneurons, support spike-timing and synaptic plasticity and are associated with perception, attention and memory. Absence of the MCU resulted in (i) gamma oscillations with decreased power (by >40%) and lower synchrony, including less precise neural action potential generation (‘spiking'), (ii) sharp waves with decreased incidence (by about 22%) and decreased fast ripple frequency (by about 3%) and (iii) lack of activity-dependent pyruvate dehydrogenase dephosphorylation. However, compensatory adaptation in gene expression related to mitochondrial function and glucose metabolism was not detected. These data suggest that the neuronal MCU is crucial for the generation of network rhythms, most likely by influences on oxidative phosphorylation and perhaps by controlling cytoplasmic Ca2+ homeostasis. This work contributes to an increased understanding of mitochondrial Ca2+ uptake in cortical information processing underlying cognition and behaviour.

Abstract 409: Investigating the Role of the Mitochondrial Calcium Uniporter (mcu) in Calcium Regulation of Cardiac Mitochondria Using Transmural Spectroscopy and an Integrating Sphere

2020 ◽  
Vol 127 (Suppl_1) ◽  
Author(s):  
Anna Kosmach ◽  
Junhui Sun ◽  
Armel Femnou ◽  
Robert S Balaban ◽  
Elizabeth Murphy
Keyword(s):  
White Light ◽  
Mitochondrial Outer Membrane ◽  
Integrating Sphere ◽  
Mitochondrial Calcium ◽  
Mouse Heart ◽  
Light Sources ◽  
Cardiac Mitochondria ◽  
Mitochondrial Calcium Uniporter ◽  
Calcium Uniporter

Cardiac mitochondria uptake calcium through the mitochondrial calcium uniporter (MCU). To better understand the role of MCU and mitochondrial calcium in regulating heart physiology and pathophysiology, we developed a method to measure mitochondrial matrix calcium in beating, perfused hearts. Langendorff perfusing hearts are loaded with 4.5 uM rhod-2-AM at 30°C and then perfused at 37°C to washout uncleaved dye. We determined that rhod-2 localized primarily to the mitochondria under our loading conditions, as shown by loading in a heart expressing a GFP-tagged mitochondrial outer membrane protein and analyzing heart slices under super resolution microscopy. Further, addition of Mn 2+ , which quenches cytosolic rhod-2, has little effect on rhod-2 signal. We insert an optical catheter with both white light and 532nm laser into the left ventricle and interleaved the collection of spectra from both light sources. The perfused heart is center mounted in an integrating sphere for spectra collection. The light passes through the ventricle and reflects off the integrating sphere resulting in a near uniform sampling of the transmitted light. Spectral properties from both light sources are determined using a rapid scanning spectrophotometer. Myoglobin oxygenation, cytochrome redox state, and rhod-2 loading are determined by white light absorbance. Ca 2+ bound rhod-2 emission is determined by removing background tissue effects from laser emission spectra and normalizing to tissue absorbance. Using this method we are able to measure changes in Ca 2+ and cytochromes during treatments such as isoproterenol and ischemia-reperfusion. The use of an integrating sphere transmural spectroscopy provides us an unique method to study mitochondrial Ca 2+ signaling in perfused mouse heart loaded with Rhod-2.

scholarly journals Mitochondrial Calcium Uniporter Deficiency in Zebrafish Causes Cardiomyopathy With Arrhythmia

2020 ◽  
Vol 11 ◽  
Author(s):  
Adam D. Langenbacher ◽  
Hirohito Shimizu ◽  
Welkin Hsu ◽  
Yali Zhao ◽  
Alexandria Borges ◽  
...  
Keyword(s):  
Energy Production ◽  
Mitochondrial Calcium ◽  
Sinus Arrest ◽  
Compact Layer ◽  
Mitochondrial Calcium Uniporter ◽  
Calcium Uniporter ◽  
Trabecular Density ◽  
And Function ◽  
Production Cell

Mitochondrial Ca2 + uptake influences energy production, cell survival, and Ca2 + signaling. The mitochondrial calcium uniporter, MCU, is the primary route for uptake of Ca2 + into the mitochondrial matrix. We have generated a zebrafish MCU mutant that survives to adulthood and exhibits dramatic cardiac phenotypes resembling cardiomyopathy and sinus arrest. MCU hearts contract weakly and have a smaller ventricle with a thin compact layer and reduced trabecular density. Damaged myofibrils and swollen mitochondria were present in the ventricles of MCU mutants, along with gene expression changes indicative of cell stress and altered cardiac structure and function. Using electrocardiography, we found that MCU hearts display conduction system defects and abnormal rhythm, with extended pauses resembling episodes of sinus arrest. Together, our findings suggest that proper mitochondrial Ca2 + homeostasis is crucial for maintaining a healthy adult heart, and establish the MCU mutant as a useful model for understanding the role of mitochondrial Ca2 + handling in adult cardiac biology.

Sign in / Sign up

Export Citation Format

Share Document