Amoeboid-like neuronal migration ensures correct horizontal cell layer formation in the developing vertebrate retina

As neurons are often born at positions different than where they ultimately function, neuronal migration is key to ensure successful nervous system development. Radial migration during which neurons featuring unipolar and bipolar morphology, employ pre-existing processes or underlying cells for directional guidance, is the most well explored neuronal migration mode. However, how neurons that display multipolar morphology, without such processes, move through highly crowded tissue environments towards their final positions remains elusive. To understand this, we here investigated multipolar migration of horizontal cells in the zebrafish retina. We found that horizontal cells tailor their movements to the environmental spatial constraints of the crowded retina, by featuring several characteristics of amoeboid migration. These include cell and nucleus shape changes, and persistent rearward polarization of stable F-actin, which enable horizontal cells to successfully move through the crowded retina. Interference with the organization of the developing retina by changing nuclear properties or overall tissue architecture, hampers efficient horizontal cell migration and layer formation. Thus, cell-tissue interplay is crucial for efficient migration of horizontal cells in the retina. In view of high proportion of multipolar neurons, the here uncovered ameboid-like neuronal migration mode might also be crucial in other areas of the developing brain.

Role of Lamin A/C as Candidate Biomarker of Aggressiveness and Tumorigenicity in Glioblastoma Multiforme

Nuclear lamina components have long been regarded as scaffolding proteins, forming a dense fibrillar structure necessary for the maintenance of the nucleus shape in all the animal kingdom. More recently, mutations, aberrant localisation and deregulation of these proteins have been linked to several diseases, including cancer. Using publicly available data we found that the increased expression levels of the nuclear protein Lamin A/C correlate with a reduced overall survival in The Cancer Genome Atlas Research Network (TCGA) patients affected by glioblastoma multiforme (GBM). We show that the expression of the LMNA gene is linked to the enrichment of cancer-related pathways, particularly pathways related to cell adhesion and cell migration. Mimicking the modulation of LMNA in a GBM preclinical cancer model, we confirmed both in vitro and in vivo that the increased expression of LMNA is associated with an increased aggressiveness and tumorigenicity. In addition, delving into the possible mechanism behind LMNA-induced GBM aggressiveness and tumorigenicity, we found that the mTORC2 component, Rictor, plays a central role in mediating these effects.

Biomarker of buccal mucosa cells damaged after exposure to panoramic radiography: a literature review

Objectives: This review aimed to understand the effect of exposure to panoramic radiographs on exfoliated buccal mucosal cells at the cellular level. Review: The dose of radiation exposure in dentistry, both intraoral and extraoral, has been regulated by The National Radiological Protection Board (NRPB). However, even though it is given in small doses, x-ray radiation due to intraoral and extraoral radiographs still has a radiobiological effect on the exposed tissue. The radiobiological effects of X-ray exposure can cause changes in biological molecules, either directly or indirectly, within hours or days. There are two classification of this radiobiological effect, called deterministic and stochastic effect. The deterministic effect occurs when the dose given exceeds the recommended dose by the NRPB, whereas the stochastic effect does not have any threshold that needs to be exceeded to give some adverse impact to the exposed tissue One method used as a predictor or biomarker of genetic damage due to exposure to physical or chemical mutagenic agents in humans is micronucleus (MN). The biomarker for the cell damaged is the change of nucleus shape and outline, called pycnosis, karyolysis, karyorrhexis. Conclusion: The exposed to x-ray from panoramic could induce cell and genetic damaged. Prescription for panoramic radiographic examination in patients should be as effectively as possible according to the principles of ALADA (as low as diagnostically acceptable) to avoid adverse effects on the exposed tissue.

The characteristics of nuclear membrane fluctuations in stem cells

We analyse the stem cell nucleus shape fluctuation spectrum obtained from optical confocal microscopy on an hour time scale with 10 s resolution. In particular, we investigate the angular and time dependencies of these fluctuations, define appropriate correlation functions that reveal the fundamentally out of equilibrium nature of the observed fluctuations as well as their global anisotropy. Langevin equations respecting the symmetry of the system allow us to model the damped oscillatory behaviour of the time correlations.

Prospects and limitations of expansion microscopy in chromatin ultrastructure determination

Expansion microscopy (ExM) is a method to magnify physically a specimen with preserved ultrastructure. It has the potential to explore structural features beyond the diffraction limit of light. The procedure has been successfully used for different animal species, from isolated macromolecular complexes through cells to tissue slices. Expansion of plant-derived samples is still at the beginning, and little is known, whether the chromatin ultrastructure becomes altered by physical expansion. In this study, we expanded isolated barley nuclei and compared whether ExM can provide a structural view of chromatin comparable with super-resolution microscopy. Different fixation and denaturation/digestion conditions were tested to maintain the chromatin ultrastructure. We achieved up to ~4.2-times physically expanded nuclei corresponding to a maximal resolution of ~50–60 nm when imaged by wild-field (WF) microscopy. By applying structured illumination microscopy (SIM, super-resolution) doubling the WF resolution, the chromatin structures were observed at a resolution of ~25–35 nm. WF microscopy showed a preserved nucleus shape and nucleoli. Moreover, we were able to detect chromatin domains, invisible in unexpanded nuclei. However, by applying SIM, we observed that the preservation of the chromatin ultrastructure after the expansion was not complete and that the majority of the tested conditions failed to keep the ultrastructure. Nevertheless, using expanded nuclei, we localized successfully centromere repeats by fluorescence in situ hybridization (FISH) and the centromere-specific histone H3 variant CENH3 by indirect immunolabelling. However, although these repeats and proteins were localized at the correct position within the nuclei (indicating a Rabl orientation), their ultrastructural arrangement was impaired.

Predicting the tension in actin cytoskeleton from the nucleus shape

Tension in actin cytoskeleton regulates many cellular processes and nuclear morphology. Here, we demonstrate a simple computational method for estimating actin cytoskeletal tension from nucleus shape. We first note that mechanics-based modeling defines a relationship among the volume, surface area, and projected area of the nucleus and hence a specific surface in the three-parameter space of the aforementioned geometric quantities. Data of nuclei from multiple cell types lie on such a surface. Furthermore, nuclei from a given cell population lie on a straight line on the surface. The location and orientation of the line varies with cell type. By using a mechanical model, we present two non-dimensional parameters, namely, the flatness and stretch indicators, which serve as curvilinear coordinates on the surface. Flatness indicator defines the extent of nuclear flattening due to actin cytoskeletal tension and the stretch indicator captures the effect of the elastic modulus of the nuclear envelope. We validate our assertions by modulating the actin cytoskeletal tension using three independent mechanisms: (i) direct downregulation by Cytochalasin D, (ii) indirect upregulation using Nocodazole, and (iii) mechanical stimulation by varying substrate stiffness. We also infer that the flatness indicator is equivalent to the ratio of the height to diameter of the nucleus and is related to the Vogel number. By using this geometric insight, we validate the predictions of our model with data from many previous studies. Finally, we present an analytical formula and a correlation for estimating actin cytoskeletal tension from nuclear projected area and volume.

Analysis of cardiomyocyte nuclei in human cardiomyopathy reveals orientation dependent defects in shape

Cardiomyopathies are progressive diseases of heart muscle often caused by mutations in genes encoding sarcomeric, cytoskeletal and nucleoskeletal proteins though in many cases the cause of disease is not identified. Whilst nucleus hypertrophy has been described, it is not known whether nucleus shape changes are a general feature of cardiomyopathy. Due to the rod-shaped nature of cardiomyocytes and their elliptical nuclei we hypothesised that orientation of analysis would be an important determinant of any changes observed between patients exhibiting primarily unexplained cardiomyopathy and control samples from non-failing donors. To investigate this we performed image analysis of cardiomyocyte nuclei in myocardial cryosections from a cohort of cardiomyopathy patients. We discovered that circularity, solidity and aspect ratio were sensitive to orientation of the myocardium and that in the transverse plane only circularity was reduced in cardiomyocyte nuclei of cardiomyopathy patients. These findings show that orientation dependent changes in nucleus shape may be a property of cardiomyopathy and with appropriate follow up studies, may prove to have mechanistic and diagnostic value.

Prospects and limitations of expansion microscopy in chromatin ultrastructure determination

Expansion Microscopy (ExM) is a method to magnify physically a specimen with preserved ultrastructure. It has the potential to explore structural features beyond the diffraction limit of light. The procedure has been successfully used for different animal species, from isolated macromolecular complexes through cells to tissue slices. Expansion of plant-derived samples is still at the beginning, and little is known whether the chromatin ultrastructure becomes altered by physical expansion.In this study, we expanded isolated barley nuclei and compared whether ExM can provide a structural view of chromatin comparable with super-resolution microscopy. Different fixation and denaturation/digestion conditions were tested to maintain the chromatin ultrastructure. We achieved up to ∼4.2-times physically expanded nuclei corresponding to a maximal resolution of ∼50-60 nm when imaged by wild-field (WF) microscopy. By applying structured illumination microscopy (SIM, super-resolution) doubling the WF resolution the chromatin structures were observed at a resolution of ∼25-35 nm.WF microscopy showed a preserved nucleus shape and nucleoli. Moreover, we were able to detect chromatin domains, invisible in unexpanded nuclei. However, by applying SIM we observed that the preservation of the chromatin ultrastructure after expansion was not complete and that the majority of the tested conditions failed to keep the ultrastructure.Nevertheless, using expanded nuclei we detected successfully centromere repeats by fluorescence in situ hybridization (FISH) and the centromere-specific histone H3 variant CENH3 by indirect immunostaining. However, although these repeats and proteins were localized at the correct position within the nuclei (indicating a Rabl orientation) their ultrastructural arrangement was impaired.

Spitzer Space Telescope observations of bilobate comet 8P/Tuttle

Context. Comet 8P/Tuttle is a nearly isotropic comet whose physical properties are poorly known and might be different from those of ecliptic comets owing to their different origin. Two independent observations have shown that 8P/Tuttle has a bilobate nucleus. Aims. Our goal is to determine the physical properties of the nucleus (size, shape, thermal inertia, and albedo) and coma (water and dust) of 8P/Tuttle. Methods. We observed the inner coma of 8P/Tuttle with the infrared spectrograph and the infrared camera of the Spitzer Space Telescope. We obtained one spectrum (5–40 μm) on 2 November 2007 and a set of 19 images at 24 μm on 22–23 June 2008 sampling the rotational period of the nucleus. The data were interpreted using thermal models for the nucleus and the dust coma, and we considered two possible shape models of the nucleus derived from Hubble Space Telescope visible and Arecibo radar observations. Results. We favor a model for the nucleus shape that is composed of two contact spheres with respective radii of 2.7 ± 0.1 km and 1.1 ± 0.1 km and a pole orientation with RA = 285 ± 12° and Dec = +20 ± 5°. The thermal inertia of the nucleus lies in the range 0–100 J K−1 m−2 s−1∕2 and the R-band geometric albedo is 0.042 ± 0.008. The water production rate amounts to 1.1 ± 0.2 × 1028 molecules s−1 at 1.6 AU from the Sun pre-perihelion, which corresponds to an active fraction of ≈9%. At the same distance, the ɛfρ quantity amounts to 310 ± 34 cm, and it reaches 325 ± 36 cm at 2.2 AU post-perihelion. The dust grain temperature is estimated to be 258 ± 10 K, which is 37 K higher than the thermal equilibrium temperature at 1.6 AU. This indicates that the dust grains that contribute to the thermal infrared flux have a typical size of ≈10 μm. The dust spectrum exhibits broad emission around 10 μm (1.5σ confidence level) and 18 μm (5σ confidence level) that we attribute to amorphous pyroxene.