scholarly journals Clock-Controlled Mitochondrial Dynamics Correlates with Cyclic Pregnenolone Synthesis

Cells ◽  
2020 ◽  
Vol 9 (10) ◽  
pp. 2323
Author(s):  
Melissa Witzig ◽  
Amandine Grimm ◽  
Karen Schmitt ◽  
Imane Lejri ◽  
Stephan Frank ◽  
...  
Keyword(s):  
Molecular Clock ◽  
Mitochondrial Dynamics ◽  
Glioma Cells ◽  
Translocator Protein ◽  
Mitochondrial Morphology ◽  
Circadian Regulation ◽  
The Core ◽  
Steroidogenic Cells ◽  
First Time ◽  
A172 Glioma Cells

Neurosteroids are steroids synthetized in the nervous system, with the first step of steroidogenesis taking place within mitochondria with the synthesis of pregnenolone. They exert important brain-specific functions by playing a role in neurotransmission, learning and memory processes, and neuroprotection. Here, we show for the first time that mitochondrial neurosteroidogenesis follows a circadian rhythm and correlates with the rhythmic changes in mitochondrial morphology. We used synchronized human A172 glioma cells, which are steroidogenic cells with a functional core molecular clock, to show that pregnenolone levels and translocator protein (TSPO) are controlled by the clock, probably via circadian regulation of mitochondrial fusion/fission. Key findings were recapitulated in mouse brains. We also showed that genetic or pharmacological abrogation of fusion/fission activity, as well as disturbing the core molecular clock, abolished circadian rhythms of pregnenolone and TSPO. Our findings provide new insights into the crosstalk between mitochondrial function (here, neurosteroidogenesis) and circadian cycles.

scholarly journals MYC-associated factor MAX is an essential regulator of the clock core network

2019 ◽  
Author(s):  
Olga Blaževitš ◽  
Nityanand Bolshette ◽  
Donatella Vecchio ◽  
Ana Guijarro ◽  
Ottavio Croci ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Molecular Clock ◽  
Circadian Regulation ◽  
Factor X ◽  
Core Network ◽  
Associated Factor ◽  
Rhythmic Expression ◽  
The Core ◽  
E Box

SummaryThe circadian transcriptional network is based on a competition between transcriptional activator and repressor complexes regulating the rhythmic expression of clock-controlled genes. We show here that the MYC-Associated factor X, MAX, plays a repressive role in this network and operates through its MYC-independent binding to E-box-containing regulatory regions within the promoters of circadian BMAL1 targets. This clock function of MAX is essential for maintaining a proper circadian rhythm but separated by the role of MAX as a partner of MYC in controlling cell proliferation. We also identified MAX Network Transcriptional repressor, MNT, as a fundamental partner of MAX-mediated circadian regulation. Collectively, our data indicate that MAX is an integral part of the core molecular clock and keeps the balance between positive and negative elements of the molecular clock machinery. Accordingly, alteration of MAX transcriptional complexes may contribute to circadian dysfunction in pathological contexts.

scholarly journals Machine learning-based classification of mitochondrial morphology in primary neurons and brain

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Garrett M. Fogo ◽  
Anthony R. Anzell ◽  
Kathleen J. Maheras ◽  
Sarita Raghunayakula ◽  
Joseph M. Wider ◽  
...  
Keyword(s):  
Image Analysis ◽  
Cortical Neurons ◽  
Mitochondrial Dynamics ◽  
Automated Image Analysis ◽  
Mitochondrial Morphology ◽  
Analysis Pipeline ◽  
Genetic Ablation ◽  
Block Face

AbstractThe mitochondrial network continually undergoes events of fission and fusion. Under physiologic conditions, the network is in equilibrium and is characterized by the presence of both elongated and punctate mitochondria. However, this balanced, homeostatic mitochondrial profile can change morphologic distribution in response to various stressors. Therefore, it is imperative to develop a method that robustly measures mitochondrial morphology with high accuracy. Here, we developed a semi-automated image analysis pipeline for the quantitation of mitochondrial morphology for both in vitro and in vivo applications. The image analysis pipeline was generated and validated utilizing images of primary cortical neurons from transgenic mice, allowing genetic ablation of key components of mitochondrial dynamics. This analysis pipeline was further extended to evaluate mitochondrial morphology in vivo through immunolabeling of brain sections as well as serial block-face scanning electron microscopy. These data demonstrate a highly specific and sensitive method that accurately classifies distinct physiological and pathological mitochondrial morphologies. Furthermore, this workflow employs the use of readily available, free open-source software designed for high throughput image processing, segmentation, and analysis that is customizable to various biological models.

scholarly journals Mitochondrial fission and mitophagy are independent mechanisms regulating ischemia/reperfusion injury in primary neurons

2021 ◽  
Vol 12 (5) ◽  
Author(s):  
Anthony R. Anzell ◽  
Garrett M. Fogo ◽  
Zoya Gurm ◽  
Sarita Raghunayakula ◽  
Joseph M. Wider ◽  
...  
Keyword(s):  
Cortical Neurons ◽  
Ischemia Reperfusion Injury ◽  
Mitochondrial Dynamics ◽  
Ischemia Reperfusion ◽  
Mitochondrial Fission ◽  
Conditional Knockout ◽  
Mitochondrial Morphology ◽  
Primary Neurons ◽  
Mitochondrial Fragmentation

AbstractMitochondrial dynamics and mitophagy are constitutive and complex systems that ensure a healthy mitochondrial network through the segregation and subsequent degradation of damaged mitochondria. Disruption of these systems can lead to mitochondrial dysfunction and has been established as a central mechanism of ischemia/reperfusion (I/R) injury. Emerging evidence suggests that mitochondrial dynamics and mitophagy are integrated systems; however, the role of this relationship in the context of I/R injury remains unclear. To investigate this concept, we utilized primary cortical neurons isolated from the novel dual-reporter mitochondrial quality control knockin mice (C57BL/6-Gt(ROSA)26Sortm1(CAG-mCherry/GFP)Ganl/J) with conditional knockout (KO) of Drp1 to investigate changes in mitochondrial dynamics and mitophagic flux during in vitro I/R injury. Mitochondrial dynamics was quantitatively measured in an unbiased manner using a machine learning mitochondrial morphology classification system, which consisted of four different classifications: network, unbranched, swollen, and punctate. Evaluation of mitochondrial morphology and mitophagic flux in primary neurons exposed to oxygen-glucose deprivation (OGD) and reoxygenation (OGD/R) revealed extensive mitochondrial fragmentation and swelling, together with a significant upregulation in mitophagic flux. Furthermore, the primary morphology of mitochondria undergoing mitophagy was classified as punctate. Colocalization using immunofluorescence as well as western blot analysis revealed that the PINK1/Parkin pathway of mitophagy was activated following OGD/R. Conditional KO of Drp1 prevented mitochondrial fragmentation and swelling following OGD/R but did not alter mitophagic flux. These data provide novel evidence that Drp1 plays a causal role in the progression of I/R injury, but mitophagy does not require Drp1-mediated mitochondrial fission.

scholarly journals When the Balance Tips: Dysregulation of Mitochondrial Dynamics as a Culprit in Disease

2021 ◽  
Vol 22 (9) ◽  
pp. 4617
Author(s):  
Styliana Kyriakoudi ◽  
Anthi Drousiotou ◽  
Petros P. Petrou
Keyword(s):  
Insulin Resistance ◽  
Mitochondrial Function ◽  
Human Disease ◽  
Molecular Mechanisms ◽  
Mitochondrial Dynamics ◽  
Mitochondrial Fusion ◽  
Mitochondrial Morphology ◽  
Disease Development ◽  
Pathological Conditions ◽  
Wide Range

Mitochondria are dynamic organelles, the morphology of which is tightly linked to their functions. The interplay between the coordinated events of fusion and fission that are collectively described as mitochondrial dynamics regulates mitochondrial morphology and adjusts mitochondrial function. Over the last few years, accruing evidence established a connection between dysregulated mitochondrial dynamics and disease development and progression. Defects in key components of the machinery mediating mitochondrial fusion and fission have been linked to a wide range of pathological conditions, such as insulin resistance and obesity, neurodegenerative diseases and cancer. Here, we provide an update on the molecular mechanisms promoting mitochondrial fusion and fission in mammals and discuss the emerging association of disturbed mitochondrial dynamics with human disease.

scholarly journals On the formation and stability of fermionic dark matter halos in a cosmological framework

2020 ◽  
Author(s):  
Carlos R Argüelles ◽  
Manuel I Díaz ◽  
Andreas Krut ◽  
Rafael Yunis
Keyword(s):  
Black Hole ◽  
Dark Matter ◽  
Space Distribution ◽  
Coarse Grained ◽  
Dark Matter Halos ◽  
The Core ◽  
The Galaxy ◽  
Gravitating Systems ◽  
The Given ◽  
First Time

Abstract The formation and stability of collisionless self-gravitating systems is a long standing problem, which dates back to the work of D. Lynden-Bell on violent relaxation, and extends to the issue of virialization of dark matter (DM) halos. An important prediction of such a relaxation process is that spherical equilibrium states can be described by a Fermi-Dirac phase-space distribution, when the extremization of a coarse-grained entropy is reached. In the case of DM fermions, the most general solution develops a degenerate compact core surrounded by a diluted halo. As shown recently, the latter is able to explain the galaxy rotation curves while the DM core can mimic the central black hole. A yet open problem is whether this kind of astrophysical core-halo configurations can form at all, and if they remain stable within cosmological timescales. We assess these issues by performing a thermodynamic stability analysis in the microcanonical ensemble for solutions with given particle number at halo virialization in a cosmological framework. For the first time we demonstrate that the above core-halo DM profiles are stable (i.e. maxima of entropy) and extremely long lived. We find the existence of a critical point at the onset of instability of the core-halo solutions, where the fermion-core collapses towards a supermassive black hole. For particle masses in the keV range, the core-collapse can only occur for Mvir ≳ E9M⊙ starting at zvir ≈ 10 in the given cosmological framework. Our results prove that DM halos with a core-halo morphology are a very plausible outcome within nonlinear stages of structure formation.

scholarly journals Nanoscopic quantification of sub-mitochondrial morphology, mitophagy and mitochondrial dynamics in living cells derived from patients with mitochondrial diseases

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Weiwei Zou ◽  
Qixin Chen ◽  
Jesse Slone ◽  
Li Yang ◽  
Xiaoting Lou ◽  
...  
Keyword(s):  
Oxidative Phosphorylation ◽  
Optic Atrophy ◽  
Respiratory Function ◽  
Clinical Symptoms ◽  
Mitochondrial Dynamics ◽  
Cerebellar Atrophy ◽  
Mitochondrial Diseases ◽  
Living Cells ◽  
Mitochondrial Morphology ◽  
Mitochondrial Oxidative Phosphorylation

AbstractSLC25A46 mutations have been found to lead to mitochondrial hyper-fusion and reduced mitochondrial respiratory function, which results in optic atrophy, cerebellar atrophy, and other clinical symptoms of mitochondrial disease. However, it is generally believed that mitochondrial fusion is attributable to increased mitochondrial oxidative phosphorylation (OXPHOS), which is inconsistent with the decreased OXPHOS of highly-fused mitochondria observed in previous studies. In this paper, we have used the live-cell nanoscope to observe and quantify the structure of mitochondrial cristae, and the behavior of mitochondria and lysosomes in patient-derived SLC25A46 mutant fibroblasts. The results show that the cristae have been markedly damaged in the mutant fibroblasts, but there is no corresponding increase in mitophagy. This study suggests that severely damaged mitochondrial cristae might be the predominant cause of reduced OXPHOS in SLC25A46 mutant fibroblasts. This study demonstrates the utility of nanoscope-based imaging for realizing the sub-mitochondrial morphology, mitophagy and mitochondrial dynamics in living cells, which may be particularly valuable for the quick evaluation of pathogenesis of mitochondrial morphological abnormalities.

scholarly journals Extended analysis of Xe VII and Xe VIII

2017 ◽  
Vol 95 (9) ◽  
pp. 805-810 ◽  
Author(s):  
M. Raineri ◽  
M. Gallardo ◽  
J. Reyna Almandos ◽  
C.J.B. Pagan ◽  
R. Sarmiento
Keyword(s):  
Light Source ◽  
Energy Levels ◽  
Excited Configuration ◽  
Pulsed Discharge ◽  
Hartree Fock ◽  
The Core ◽  
New Energy ◽  
Extended Analysis ◽  
First Time ◽  
Atomic Parameters

A pulsed discharge light source to study the six and seven times ionized xenon spectra in the 419–4642 Å region was used. A set of 40 transitions of Xe VII and 25 transitions of Xe VIII were classified for the first time. We revised the values for the previously known energy levels and extended the analysis for Xe VII to 10 new energy levels belonging to 5s6d, 5s7s and 5s7p, 4d95s25p even and odd configurations, respectively. Seven new energy levels of the core excited configuration 4d95s5d of Xe VIII are presented. For the prediction of the atomic parameters, energy levels, and transition, relativistic Hartree–Fock calculations were used.

scholarly journals High glucose induces Drp1-mediated mitochondrial fission via the Orai1 calcium channel to participate in diabetic cardiomyocyte hypertrophy

2021 ◽  
Vol 12 (2) ◽  
Author(s):  
Qing-Rui Wu ◽  
Dan-Lin Zheng ◽  
Pei-Ming Liu ◽  
Hui Yang ◽  
Lu-An Li ◽  
...  
Keyword(s):  
Calcium Channel ◽  
Mitochondrial Dysfunction ◽  
High Glucose ◽  
Mitochondrial Dynamics ◽  
Mitochondrial Fission ◽  
Calcium Handling ◽  
Cardiomyocyte Hypertrophy ◽  
Mitochondrial Morphology ◽  
Calmodulin Binding ◽  
Downstream Target

AbstractMitochondrial dysfunction and impaired Ca2+ handling are involved in the development of diabetic cardiomyopathy (DCM). Dynamic relative protein 1 (Drp1) regulates mitochondrial fission by changing its level of phosphorylation, and the Orai1 (Ca2+ release-activated calcium channel protein 1) calcium channel is important for the increase in Ca2+ entry into cardiomyocytes. We aimed to explore the mechanism of Drp1 and Orai1 in cardiomyocyte hypertrophy caused by high glucose (HG). We found that Zucker diabetic fat rats induced by administration of a high-fat diet develop cardiac hypertrophy and impaired cardiac function, accompanied by the activation of mitochondrial dynamics and calcium handling pathway-related proteins. Moreover, HG induces cardiomyocyte hypertrophy, accompanied by abnormal mitochondrial morphology and function, and increased Orai1-mediated Ca2+ influx. Mechanistically, the Drp1 inhibitor mitochondrial division inhibitor 1 (Mdivi-1) prevents cardiomyocyte hypertrophy induced by HG by reducing phosphorylation of Drp1 at serine 616 (S616) and increasing phosphorylation at S637. Inhibition of Orai1 with single guide RNA (sgOrai1) or an inhibitor (BTP2) not only suppressed Drp1 activity and calmodulin-binding catalytic subunit A (CnA) and phosphorylated-extracellular signal-regulated kinase (p-ERK1/2) expression but also alleviated mitochondrial dysfunction and cardiomyocyte hypertrophy caused by HG. In addition, the CnA inhibitor cyclosporin A and p-ERK1/2 inhibitor U0126 improved HG-induced cardiomyocyte hypertrophy by promoting and inhibiting phosphorylation of Drp1 at S637 and S616, respectively. In summary, we identified Drp1 as a downstream target of Orai1-mediated Ca2+ entry, via activation by p-ERK1/2-mediated phosphorylation at S616 or CnA-mediated dephosphorylation at S637 in DCM. Thus, the Orai1–Drp1 axis is a novel target for treating DCM.

Measurements and Modelling of Ingress in a New 1.5-Stage Turbine Research Facility

2016 ◽  
Author(s):  
Marios Patinios ◽  
James A. Scobie ◽  
Carl M. Sangan ◽  
J. Michael Owen ◽  
Gary D. Lock
Keyword(s):  
Theoretical Model ◽  
Gas Turbines ◽  
Test Facility ◽  
Radial Clearance ◽  
Operating Life ◽  
The Core ◽  
Wide Range ◽  
Purge Flow ◽  
Engine Components ◽  
First Time

In gas turbines, hot mainstream flow can be ingested into the wheel-space formed between stator and rotor discs as a result of the circumferential pressure asymmetry in the annulus; this ingress can significantly affect the operating life, performance and integrity of highly-stressed, vulnerable engine components. Rim seals, fitted at the periphery of the discs, are used to minimise ingress and therefore reduce the amount of purge flow required to seal the wheel-space and cool the discs. This paper presents experimental results from a new 1.5-stage test facility designed to investigate ingress into the wheel-spaces upstream and downstream of a rotor disc. The fluid-dynamically-scaled rig operates at incompressible flow conditions, far removed from the harsh environment of the engine which is not conducive to experimental measurements. The test facility features interchangeable rim-seal components, offering significant flexibility and expediency in terms of data collection over a wide range of sealing-flow rates. The rig was specifically designed to enable an efficient method of ranking and quantifying the performance of generic and engine-specific seal geometries. The radial variation of CO2 gas concentration, pressure and swirl is measured to explore, for the first time, the flow structure in both the upstream and downstream wheel-spaces. The measurements show that the concentration in the core is equal to that on the stator walls and that both distributions are virtually invariant with radius. These measurements confirm that mixing between ingress and egress is essentially complete immediately after the ingested fluid enters the wheel-space and that the fluid from the boundary-layer on the stator is the source of that in the core. The swirl in the core is shown to determine the radial distribution of pressure in the wheel-space. The performance of a double radial-clearance seal is evaluated in terms of the variation of effectiveness with sealing flow rate for both the upstream and the downstream wheel-spaces and is found to be independent of rotational Reynolds number. A simple theoretical orifice model was fitted to the experimental data showing good agreement between theory and experiment for all cases. This observation is of great significance as it demonstrates that the theoretical model can accurately predict ingress even when it is driven by the complex unsteady pressure field in the annulus upstream and downstream of the rotor. The combination of the theoretical model and the new test rig with its flexibility and capability for detailed measurements provides a powerful tool for the engine rim-seal designer.

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