5,408 research outputs found
Super-resolution microscopy of mitochondria.
Mitochondria, the powerhouses of the cell, are essential organelles in eukaryotic cells. With their complex inner architecture featuring a smooth outer and a highly convoluted inner membrane, they are challenging objects for microscopy. The diameter of mitochondria is generally close to the resolution limit of conventional light microscopy, rendering diffraction-unlimited super-resolution light microscopy (nanoscopy) for imaging submitochondrial protein distributions often mandatory. In this review, we discuss what can be expected when imaging mitochondria with conventional diffraction-limited and diffraction-unlimited microscopy. We provide an overview on recent studies using super-resolution microscopy to investigate mitochondria and discuss further developments and challenges in mitochondrial biology that might by addressed with these technologies in the future
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The Time Course of Gene Expression during Reactive Gliosis in the Optic Nerve
Reactive gliosis is a complex process that involves changes in gene expression and morphological remodeling. The mouse optic nerve, where astrocytes, microglia and oligodendrocytes interact with retinal ganglion cell axons and each other, is a particularly suitable model for studying the molecular mechanisms of reactive gliosis. We triggered gliosis at the mouse optic nerve head by retro orbital nerve crush. We followed the expression profiles of 14,000 genes from 1 day to 3 months, as the optic nerve formed a glial scar. The transcriptome showed profound changes. These were greatest shortly after injury; the numbers of differentially regulated genes then dropped, returning nearly to resting levels by 3 months. Different genes were modulated with very different time courses, and functionally distinct groups of genes responded in partially overlapping waves. These correspond roughly to two quick waves of inflammation and cell proliferation, a slow wave of tissue remodeling and debris removal, and a final stationary phase that primarily reflects permanent structural changes in the axons. Responses from astrocytes, microglia and oligodendrocytes were distinctively different, both molecularly and morphologically. Comparisons to other models of brain injury and to glaucoma indicated that the glial responses depended on both the tissue and the injury. Attempts to modulate glial function after axonal injuries should consider different mechanistic targets at different times following the insult
The TFAM-to-mtDNA ratio defines inner-cellular nucleoid populations with distinct activity levels
In human cells, generally a single mitochondrial DNA (mtDNA) is compacted into a nucleoprotein complex denoted the nucleoid. Each cell contains hundreds of nucleoids, which tend to cluster into small groups. It is unknown whether all nucleoids are equally involved in mtDNA replication and transcription or whether distinct nucleoid subpopulations exist. Here, we use multi-color STED super-resolution microscopy to determine the activity of individual nucleoids in primary human cells. We demonstrate that only a minority of all nucleoids are active. Active nucleoids are physically larger and tend to be involved in both replication and transcription. Inactivity correlates with a high ratio of the mitochondrial transcription factor A (TFAM) to the mtDNA of the individual nucleoid, suggesting that TFAM-induced nucleoid compaction regulates nucleoid replication and transcription activity in vivo. We propose that the stable population of highly compacted inactive nucleoids represents a storage pool of mtDNAs with a lower mutational load
A renormalization group approach to time dependent transport through correlated quantum dots
We introduce a real time version of the functional renormalization group
which allows to study correlation effects on nonequilibrium transport through
quantum dots. Our method is equally capable to address (i) the relaxation out
of a nonequilibrium initial state into a (potentially) steady state driven by a
bias voltage and (ii) the dynamics governed by an explicitly time-dependent
Hamiltonian. All time regimes from transient to asymptotic can be tackled; the
only approximation is the consistent truncation of the flow equations at a
given order. As an application we investigate the relaxation dynamics of the
interacting resonant level model which describes a fermionic quantum dot
dominated by charge fluctuations. Moreover, we study decoherence and relaxation
phenomena within the ohmic spin-boson model by mapping the latter to the
interacting resonant level model
Mitochondrial protein abundance gradients require the distribution of separated mitochondria
Mitochondria are highly dynamic organelles that interchange their contents mediated by fission and fusion. However, it has previously been shown that the mitochondria of cultured human epithelial cells exhibit a gradient in the relative abundance of several proteins, with the perinuclear mitochondria generally exhibiting a higher protein abundance than the peripheral mitochondria. The molecular mechanisms that are required for the establishment and the maintenance of such inner-cellular mitochondrial protein abundance gradients are unknown. We verified the existence of inner-cellular gradients in the abundance of clusters of the mitochondrial outer membrane protein Tom20 in the mitochondria of kidney epithelial cells from an African green monkey (Vero cells) using STED nanoscopy and confocal microscopy. We found that the Tom20 gradients are established immediately after cell division and require the presence of microtubules. Furthermore, the gradients are abrogated in hyperfused mitochondrial networks. Our results suggest that inner-cellular protein abundance gradients from the perinuclear to the peripheral mitochondria are established by the trafficking of individual mitochondria to their respective cellular destination
Mitochondrial cristae revealed with focused light.
Because of the diffraction resolution barrier, optical microscopes have so far failed in visualizing the mitochondrial cristae, that is, the folds of the inner membrane of this 200 to 400 nm diameter sized tubular organelle. Realizing a ∼30 nm isotropic subdiffraction resolution in isoSTED fluorescence nanoscopy, we have visualized these essential structures in the mitochondria of intact cells. We find a pronounced heterogeneity in the cristae arrangements even within individual mitochondrial tubules
Prospects for the Search for a Standard Model Higgs Boson in ATLAS using Vector Boson Fusion
The potential for the discovery of a Standard Model Higgs boson in the mass
range m_H < 2 m_Z in the vector boson fusion mode has been studied for the
ATLAS experiment at the LHC. The characteristic signatures of additional jets
in the forward regions of the detector and of low jet activity in the central
region allow for an efficient background rejection. Analyses for the H -> WW
and H -> tau tau decay modes have been performed using a realistic simulation
of the expected detector performance. The results obtained demonstrate the
large discovery potential in the H -> WW decay channel and the sensitivity to
Higgs boson decays into tau-pairs in the low-mass region around 120 GeV.Comment: 20 pages, 13 ps figures, uses EPJ style fil
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