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Neural Regeneration Research Apr 2023The mitochondrial permeability transition pore is a nonspecific transmembrane channel. Inhibition of mitochondrial permeability transition pore opening has been shown to...
The mitochondrial permeability transition pore is a nonspecific transmembrane channel. Inhibition of mitochondrial permeability transition pore opening has been shown to alleviate mitochondrial swelling, calcium overload, and axonal degeneration. Cyclophilin D is an important component of the mitochondrial permeability transition pore. Whether cyclophilin D participates in mitochondrial impairment and axonal injury after intracerebral hemorrhage is not clear. In this study, we established mouse models of intracerebral hemorrhage in vivo by injection of autologous blood and oxyhemoglobin into the striatum in Thy1-YFP mice, in which pyramidal neurons and axons express yellow fluorescent protein. We also simulated intracerebral hemorrhage in vitro in PC12 cells using oxyhemoglobin. We found that axonal degeneration in the early stage of intracerebral hemorrhage depended on mitochondrial swelling induced by cyclophilin D activation and mitochondrial permeability transition pore opening. We further investigated the mechanism underlying the role of cyclophilin D in mouse models and PC12 cell models of intracerebral hemorrhage. We found that both cyclosporin A inhibition and short hairpin RNA interference of cyclophilin D reduced mitochondrial permeability transition pore opening and mitochondrial injury. In addition, inhibition of cyclophilin D and mitochondrial permeability transition pore opening protected corticospinal tract integrity and alleviated motor dysfunction caused by intracerebral hemorrhage. Our findings suggest that cyclophilin D is used as a key mediator of axonal degeneration after intracerebral hemorrhage; inhibition of cyclophilin D expression can protect mitochondrial structure and function and further alleviate corticospinal tract injury and motor dysfunction after intracerebral hemorrhage. Our findings provide a therapeutic target for preventing axonal degeneration of white matter injury and subsequent functional impairment in central nervous diseases.
PubMed: 36204853
DOI: 10.4103/1673-5374.353495 -
Liver Research Dec 2019Liver injury and acute liver failure caused by acetaminophen (APAP) overdose is the clinically most important drug toxicity in western countries. Mechanistic...
Liver injury and acute liver failure caused by acetaminophen (APAP) overdose is the clinically most important drug toxicity in western countries. Mechanistic investigations have revealed a central role of mitochondria in the pathophysiology. Excess formation of the reactive metabolite N-acetyl-p-benzoquinone imine (NAPQI) after an overdose leads to hepatic glutathione depletion, mitochondrial protein adducts formation and an initial oxidant stress, which triggers the activation of mitogen activated protein (MAP) kinase cascade ultimately leading to c-jun N-terminal kinase (JNK) phosphorylation. Phospho-JNK translocates to the mitochondria and amplifies the oxidative and nitrosative stress eventually causing the mitochondrial membrane permeability transition pore opening and cessation of ATP synthesis. In addition, mitochondrial matrix swelling ruptures the outer membrane and releases endonucleases, which cause nuclear DNA fragmentation. Together, the nuclear DNA damage and the extensive mitochondrial dysfunction result in necrotic cell death. However, the pro-cell death signaling events are counteracted by adaptive responses such as autophagy and mitochondrial biogenesis. The improved mechanistic insight into the pathophysiology leads to better understanding of the mechanisms of action of the existing antidote N-acetylcysteine and justifies the clinical testing of novel therapeutics such as 4-methylpyrazole and calmangafodipir.
PubMed: 32655976
DOI: 10.1016/j.livres.2019.10.002 -
PLoS Genetics Sep 2021Dopamine (DA) neurons of the midbrain are at risk to become affected by mitochondrial damage over time and mitochondrial defects have been frequently reported in...
Dopamine (DA) neurons of the midbrain are at risk to become affected by mitochondrial damage over time and mitochondrial defects have been frequently reported in Parkinson's disease (PD) patients. However, the causal contribution of adult-onset mitochondrial dysfunction to PD remains uncertain. Here, we developed a mouse model lacking Mitofusin 2 (MFN2), a key regulator of mitochondrial network homeostasis, in adult midbrain DA neurons. The knockout mice develop severe and progressive DA neuron-specific mitochondrial dysfunction resulting in neurodegeneration and parkinsonism. To gain further insights into pathophysiological events, we performed transcriptomic analyses of isolated DA neurons and found that mitochondrial dysfunction triggers an early onset immune response, which precedes mitochondrial swelling, mtDNA depletion, respiratory chain deficiency and cell death. Our experiments show that the immune response is an early pathological event when mitochondrial dysfunction is induced in adult midbrain DA neurons and that neuronal death may be promoted non-cell autonomously by the cross-talk and activation of surrounding glial cells.
Topics: Animals; DNA, Mitochondrial; Disease Models, Animal; Dopaminergic Neurons; Homeostasis; Immunity; Mesencephalon; Mice; Mitochondria; Parkinsonian Disorders
PubMed: 34570766
DOI: 10.1371/journal.pgen.1009822 -
Food Chemistry: X Oct 2023This study aimed to investigate how postmortem muscle cells' mitochondria changed in morphology from three aspects: the outer membrane, cristae, and fission/fusion....
This study aimed to investigate how postmortem muscle cells' mitochondria changed in morphology from three aspects: the outer membrane, cristae, and fission/fusion. Atomic force microscopy (AFM) results showed that mitochondria underwent a morphology transformation from normal to swelling and collapse. Meanwhile, the cleavage of OPA1, upregulation of OMA1, downregulation of Mic60 and transmission electron microscope micrographs revealed that mitochondrial cristae ruptured with an aging time extended. Additionally, the increased expressions of Fis1 and Drp1, and the AFM topographic images mutually confirmed mitochondrial fission. These results further proved from the perspective of mitochondrial morphology that the degree of mitochondrial damage increased with the postmortem aging time extended, which was consistent with the results of the release of cytochrome caused by the increase of mitochondrial permeability transition pore opening and the decrease of mitochondrial membrane permeability, and further induced the apoptosis of postmortem muscle cells.
PubMed: 37780314
DOI: 10.1016/j.fochx.2023.100806 -
World Journal of Gastroenterology Apr 2023Ferroptosis is an emerging novel form of non-apoptotic, regulated cell death that is heavily dependent on iron and characterized by rupture in plasma membrane.... (Review)
Review
Ferroptosis is an emerging novel form of non-apoptotic, regulated cell death that is heavily dependent on iron and characterized by rupture in plasma membrane. Ferroptosis is distinct from other regulated cell death modalities at the biochemical, morphological, and molecular levels. The ferroptotic signature includes high membrane density, cytoplasmic swelling, condensed mitochondrial membrane, and outer mitochondrial rupture with associated features of accumulation of reactive oxygen species and lipid peroxidation. The selenoenzyme glutathione peroxidase 4, a key regulator of ferroptosis, greatly reduces the lipid overload and protects the cell membrane against oxidative damage. Ferroptosis exerts a momentous role in regulating cancer signaling pathways and serves as a therapeutic target in cancers. Dysregulated ferroptosis orchestrates gastrointestinal (GI) cancer signaling pathways leading to GI tumors such as colonic cancer, pancreatic cancer, and hepatocellular carcinoma. Crosstalk exists between ferroptosis and other cell death modalities. While apoptosis and autophagy play a detrimental role in tumor progression, depending upon the factors associated with tumor microenvironment, ferroptosis plays a decisive role in either promoting tumor growth or suppressing it. Several transcription factors, such as , activating transcription factors 3 and 4, are involved in influencing ferroptosis. Importantly, several molecular mediators of ferroptosis, such as p53, nuclear factor erythroid 2-related factor 2/heme oxygenase-1, hypoxia inducible factor 1, and sirtuins, coordinate with ferroptosis in GI cancers. In this review, we elaborated on key molecular mechanisms of ferroptosis and the signaling pathways that connect ferroptosis to GI tumors.
Topics: Humans; Ferroptosis; Carcinoma, Hepatocellular; Gastrointestinal Neoplasms; Lipid Peroxidation; Reactive Oxygen Species; Signal Transduction; Liver Neoplasms; Tumor Microenvironment
PubMed: 37179581
DOI: 10.3748/wjg.v29.i16.2433 -
International Journal of Molecular... Aug 2021The opening of the permeability transition pore (mPTP) in mitochondria initiates cell death in numerous diseases. The regulation of mPTP by NAD(H) in the mitochondrial...
The opening of the permeability transition pore (mPTP) in mitochondria initiates cell death in numerous diseases. The regulation of mPTP by NAD(H) in the mitochondrial matrix is well established; however, the role of extramitochondrial (cytosolic) NAD(H) is still unclear. We studied the effect of added NADH and NAD on: (1) the Ca-retention capacity (CRC) of isolated rat liver, heart, and brain mitochondria; (2) the Ca-dependent mitochondrial swelling in media whose particles can (KCl) or cannot (sucrose) be extruded from the matrix by mitochondrial carriers; (3) the Ca-dependent mitochondrial depolarization and the release of entrapped calcein from mitochondria of permeabilized hepatocytes; and (4) the Ca-dependent mitochondrial depolarization and subsequent repolarization. NADH and NAD increased the CRC of liver, heart, and brain mitochondria 1.5-2.5 times, insignificantly affecting the rate of Ca-uptake and the free Ca concentration in the medium. NAD(H) suppressed the Ca-dependent mitochondrial swelling both in KCl- and sucrose-based media but did not induce the contraction and repolarization of swollen mitochondria. By contrast, EGTA caused mitochondrial repolarization in both media and the contraction in KCl-based medium only. NAD(H) delayed the Ca-dependent depolarization and the release of calcein from individual mitochondria in hepatocytes. These data unambiguously demonstrate the existence of an external NAD(H)-dependent site of mPTP regulation.
Topics: Animals; Calcium; Fluoresceins; Hepatocytes; Male; Mitochondria, Heart; Mitochondria, Liver; Mitochondrial Permeability Transition Pore; NAD; Rats; Rats, Wistar
PubMed: 34445270
DOI: 10.3390/ijms22168560 -
Scientific Reports Sep 2021Vertebrate photoreceptors contain large numbers of closely-packed mitochondria which sustain the high metabolic demands of these cells. These mitochondria populations...
Vertebrate photoreceptors contain large numbers of closely-packed mitochondria which sustain the high metabolic demands of these cells. These mitochondria populations are dynamic and undergo fusion and fission events. This activity serves to maintain the population in a healthy state. In the event of mitochondrial damage, sub-domains, or indeed whole mitochondria, can be degraded and population homeostasis achieved. If this process is overwhelmed cell death may result. Death of photoreceptors contributes to loss of vision in aging individuals and is associated with many eye diseases. In this study we used serial block face scanning electron microscopy of adult Macaca fascicularis retinae to examine the 3D structure of mitochondria in rod and cone photoreceptors. We show that healthy-looking photoreceptors contain mitochondria exhibiting a range of shapes which are associated with different regions of the cell. In some photoreceptors we observe mitochondrial swelling and other changes often associated with cellular stress. In rods and cones that appear stressed we identify elongated domains of mitochondria with densely-packed normal cristae associated with photoreceptor ciliary rootlet bundles. We observe mitochondrial fission and mitochondrion fragments localised to these domains. Swollen mitochondria with few intact cristae are located towards the periphery of the photoreceptor inner-segment in rods, whilst they are found throughout the cell in cones. Swollen mitochondria exhibit sites on the mitochondrial inner membrane which have undergone complex invagination resulting in membranous, electron-dense aggregates. Membrane contact occurs between the mitochondrion and the photoreceptor plasma membrane in the vicinity of these aggregates, and a series of subsequent membrane fusions results in expulsion of the mitochondrial aggregate from the photoreceptor. These events are primarily associated with rods. The potential fate of this purged material and consequences of its clearance by retinal pigment epithelia are discussed.
Topics: Animals; Cell Membrane; Imaging, Three-Dimensional; Macaca fascicularis; Microscopy, Electron, Scanning; Mitochondria; Mitochondrial Membranes; Retinal Cone Photoreceptor Cells; Retinal Rod Photoreceptor Cells
PubMed: 34552195
DOI: 10.1038/s41598-021-98409-7 -
Journal of Personalized Medicine Jan 2022Tetracycline-3 (4-dedimethylamino sancycline, COL-3) is a non-antibiotic tetracycline derivative. COL-3 exerts potent anti-metalloproteinase activity and its antitumor...
Tetracycline-3 (4-dedimethylamino sancycline, COL-3) is a non-antibiotic tetracycline derivative. COL-3 exerts potent anti-metalloproteinase activity and its antitumor effects have been reported both in vitro and in vivo. In this study, we investigated the mechanisms of COL-3-induced cytotoxicity in a chronic myeloid leukemia cell line, K562, characterized by the BCR-ABL fusion protein. COL-3 induced K562 cell death in a concentration-dependent manner with an IC50 of 10.8 µg/mL and exhibited features of both apoptosis and necrosis. However, flow cytometry analysis revealed that necrotic cells dominated over the early and late apoptotic cells upon treatment with COL-3. Transmission electron microscopy analysis in combination with Western blotting (WB) analysis revealed early mitochondrial swelling accompanied by the early release of cytochrome c and truncated apoptosis inducing factor (tAIF). In addition, ultrastructural changes were detected in the endoplasmic reticulum (ER). COL-3 affected the levels of glucose-regulated protein-94 (GRP94) and resulted in m-calpain activation. DNA double strand breaks as a signature for DNA damage was also confirmed using an antibody against γH2AX. WB analyses did not demonstrate caspase activation, while Bcl-xL protein remained unaffected. In conclusion, COL-3-induced cell death involves DNA damage as well as mitochondrial and ER perturbation with features of paraptosis and programmed necrosis.
PubMed: 35055357
DOI: 10.3390/jpm12010042 -
ZooKeys 2022Oil beetles are meloids, which are characterised for their cleptoparasitic habits in bee nests and oily fluid of cantharidin that causes blistering and swelling of the...
Oil beetles are meloids, which are characterised for their cleptoparasitic habits in bee nests and oily fluid of cantharidin that causes blistering and swelling of the skin. The complete mitochondrial genome of is determined using the next-generation sequencing technology and its genomic characteristics are described. The 15,653-bp long genome is a circular molecule consisting of 13 protein-coding genes (PCG), 22 transport RNA, two ribosomal RNA, and a control region. The A + T bias of the mitochondrial genome is manifested in the complete sequence and the codon usage of protein-coding genes. The genetic distance within and between genera is calculated to confirm the taxonomic status of . The phylogenetic relationships among 15 available meloid taxa are inferred by the maximum likelihood (ML) method based on 13 mitochondrial PCGs. The ML trees resulting from nucleotide and amino acid datasets recover both the monophyly of and and the polyphyly comprising and . This study provides the first description of a mitochondrial genome belonging to the genus . The mitochondrial genome sequence and its characteristics are expected to be conducive to future studies on taxonomy, systematics, and molecular phylogenetics of the family Meloidae.
PubMed: 36762340
DOI: 10.3897/zookeys.1109.81544 -
Molecules (Basel, Switzerland) Dec 2023The aim of this study was to investigate the effects of sodium salicylate (SS) on the preservation and metabolic regulation of sheep sperm. Under 4 °C low-temperature...
The aim of this study was to investigate the effects of sodium salicylate (SS) on the preservation and metabolic regulation of sheep sperm. Under 4 °C low-temperature conditions, SS (at 10 µM, 20 µM, 30 µM, and 50 µM) was added to the semen diluent to detect sperm motility, plasma membrane, and acrosome integrity. Based on the selected optimal concentration of SS (20 µM), the effects of 20 µM of SS on sperms' antioxidant capacity and mitochondrial membrane potential (MMP) were evaluated, and metabolomics analysis was conducted. The results showed that on the 20th day of low-temperature storage, the sperm motility of the 20 µM SS group was 62.80%, and the activities of catalase (CAT) and superoxide dismutase (SOD) were significantly higher than those of the control group ( < 0.01). The content of Ca, reactive oxygen species (ROS), and malondialdehyde (MDA) were significantly lower than those of the control group ( < 0.01), and the total antioxidant capacity (T-AOC) was significantly higher than that of the control group ( < 0.05); mitochondrial activity and the total cholesterol (TC) content were significantly higher than those in the control group ( < 0.01). An ultrastructural examination showed that in the SS group, the sperm plasma membrane and acrosome were intact, the fibrous sheath and axoneme morphology of the outer dense fibers were normal, and the mitochondria were arranged neatly. In the control group, there was significant swelling of the sperm plasma membrane, rupture of the acrosome, and vacuolization of mitochondria. Using metabolomics analysis, 20 of the most significant differential metabolic markers were screened, mainly involving 6 metabolic pathways, with the amino acid biosynthesis pathway being the most abundant. In summary, 20 µM of SS significantly improved the preservation quality of sheep sperm under low-temperature conditions of 4 °C.
Topics: Male; Animals; Sheep; Sodium Salicylate; Semen; Antioxidants; Sperm Motility; Spermatozoa
PubMed: 38202772
DOI: 10.3390/molecules29010188