The Parkinson-related E193K LRRK2 variant impacts neuronal vesicles dynamics through perturbed protein interactions

The Leucine-Rich Repeat Kinase 2 (LRRK2) is a complex protein, expressed in neurons and implicated in Parkinson disease (PD). LRRK2 contains a dual enzymatic activity and several structural domains that constitute a versatile platform for multiple protein interactions at the synapses. In this study, we characterize the functional role of the N-terminal Armadillo repeats domain of LRRK2 and the impact on synaptic vesicle (SV) dynamics of a novel variant, E193K, harboured within this domain and identified in an Italian family affected by PD. Using a genetically encoded sensor of recycling, synaptopHluorine, and total internal reflection fluorescence microscopy, we visualized SV trafficking in the N2A neuroblastoma cells expressing the wild type LRRK2 protein, a mutant lacking the Armadillo domain (\u394N LRRK2) or the E193K variant. We found that expression of the \u394N construct increased the frequency and the amplitude of spontaneous synaptic events. A similar phenotype was detected in the presence of the E193K variant, suggesting that this mutation behaves as a loss-of-function mutation. A domain-based pulldown approach demonstrated that the LRRK2 N-terminus binds to cytoskeletal (\u3b2-actin and \u3b1-tubulin) and SV (synapsin I) proteins and the E193K substitution alters strength and quality of LRRK2 interactions. The results support a role of the Armadillo domain in interaction with synaptic proteins and suggest that the E193K mutation affects LRRK2 function via perturbation of its physiological network of interactors, resulting in impaired vesicular trafficking. These findings may have important implications for understanding the role of LRRK2 at the synapses and the pathophysiological mechanism for LRRK2-linked disease

Anomalies in thickness measurements of graphene and few layer graphite crystals by tapping mode atomic force microscopy

Atomic Force Microscopy (AFM) in the tapping (intermittent contact) mode is a commonly used tool to measure the thickness of graphene and few layer graphene (FLG) flakes on silicon oxide surfaces. It is a convenient tool to quickly determine the thickness of individual FLG films. However, reports from literature show a large variation of the measured thickness of graphene layers. This paper is focused on the imaging mechanism of tapping mode AFM (TAFM) when measuring graphene and FLG thickness and we show that at certain measurement parameters significant deviations can be introduced in the measured thickness of FLG flakes. An increase of as much as 1 nm can be observed in the measured height of FLG crystallites, when using an improperly chosen range of free amplitude values of the tapping cantilever. We present comparative Raman spectroscopy and TAFM measurements on selected single and multilayer graphene films, based on which we suggest ways to correctly measure graphene and FLG thickness using TAFM

In-situ probing of the thermal treatment of h-BN towards exfoliation

Two-dimensional (2D) hexagonal boron nitride (h-BN) is becoming increasingly interesting for wider engineering applications. Thermal exfoliation is being suggested as a facile technology to produce large quantities of 2D h-BN. Further optimization of the process requires fundamental understanding of the exfoliation mechanism, which is hardly realized by ex-situ techniques. In this study, time resolved in-situ synchrotron X-ray powder diffraction experiments are conducted while heat treating bulk h-BN up to 1273 K. During the heating process, linear expansion of c-axis is observed and the contraction of a-axis up to around 750 K is consistent with previous research. However, a changing behavior from contraction to expansion in a- axis direction is newly observed when heating over 750 K. With the consideration of previous thermally oxidation studies, a hypothesis of thermal assisted exfoliation with oxygen interstitial and substitution of nitrogen at high temperature is proposed

Raman spectroscopy of graphene and bilayer under biaxial strain: bubbles and balloons

In this letter we use graphene bubbles to study the Raman spectrum of graphene under biaxial (e.g. isotropic) strain. Our Gruneisen parameters are in excellent agreement with the theoretical values. Discrepancy in the previously reported values is attributed to the interaction of graphene with the substrate. Bilayer balloons (intentionally pressurized membranes) have been used to avoid the effect of the substrate and to study the dependence of strain on the inter-layer interactions.Comment: 14 pages, 4 figure

Probing Mechanical Properties of Graphene with Raman Spectroscopy

The use of Raman scattering techniques to study the mechanical properties of graphene films is reviewed here. The determination of Gruneisen parameters of suspended graphene sheets under uni- and bi-axial strain is discussed and the values are compared to theoretical predictions. The effects of the graphene-substrate interaction on strain and to the temperature evolution of the graphene Raman spectra are discussed. Finally, the relation between mechanical and thermal properties is presented along with the characterization of thermal properties of graphene with Raman spectroscopy.Comment: To appear in the Journal of Materials Scienc

Graphene Photonics and Optoelectronics

The richness of optical and electronic properties of graphene attracts enormous interest. Graphene has high mobility and optical transparency, in addition to flexibility, robustness and environmental stability. So far, the main focus has been on fundamental physics and electronic devices. However, we believe its true potential to be in photonics and optoelectronics, where the combination of its unique optical and electronic properties can be fully exploited, even in the absence of a bandgap, and the linear dispersion of the Dirac electrons enables ultra-wide-band tunability. The rise of graphene in photonics and optoelectronics is shown by several recent results, ranging from solar cells and light emitting devices, to touch screens, photodetectors and ultrafast lasers. Here we review the state of the art in this emerging field.Comment: Review Nature Photonics, in pres

Raman spectra of epitaxial graphene on SiC and of epitaxial graphene transferred to SiO2

Raman spectra were measured for mono-, bi- and trilayer graphene grown on SiC by solid state graphitization, whereby the number of layers was pre-assigned by angle-resolved ultraviolet photoemission spectroscopy. It was found that the only unambiguous fingerprint in Raman spectroscopy to identify the number of layers for graphene on SiC(0001) is the linewidth of the 2D (or D*) peak. The Raman spectra of epitaxial graphene show significant differences as compared to micromechanically cleaved graphene obtained from highly oriented pyrolytic graphite crystals. The G peak is found to be blue-shifted. The 2D peak does not exhibit any obvious shoulder structures but it is much broader and almost resembles a single-peak even for multilayers. Flakes of epitaxial graphene were transferred from SiC onto SiO2 for further Raman studies. A comparison of the Raman data obtained for graphene on SiC with data for epitaxial graphene transferred to SiO2 reveals that the G peak blue-shift is clearly due to the SiC substrate. The broadened 2D peak however stems from the graphene structure itself and not from the substrate.Comment: 27 pages, 8 figure

First-principles modeling of the polycyclic aromatic hydrocarbons reduction

Density functional theory modelling of the reduction of realistic nanographene molecules (C42H18, C48H18 and C60H24) by molecular hydrogen evidences for the presence of limits in the hydrogenation process. These limits caused the contentions between three-fold symmetry of polycyclic aromatic hydrocarbon molecules and two-fold symmetry of adsorbed hydrogen pairs. Increase of the binding energy between nanographenes during reduction is also discussed as possible cause of the experimentally observed limited hydrogenation of studied nanographenes.Comment: 18 pages, 7 figures, accepted to J. Phys. Chem.

Anomalous Lattice Vibrations of Single and Few-Layer MoS2

Molybdenum disulfide (MoS2) of single and few-layer thickness was exfoliated on SiO2/Si substrate and characterized by Raman spectroscopy. The number of S-Mo-S layers of the samples was independently determined by contact-mode atomic-force microscopy. Two Raman modes, E12g and A1g, exhibited sensitive thickness dependence, with the frequency of the former decreasing and that of the latter increasing with thickness. The results provide a convenient and reliable means for determining layer thickness with atomic-level precision. The opposite direction of the frequency shifts, which cannot be explained solely by van der Waals interlayer coupling, is attributed to Coulombic interactions and possible stacking-induced changes of the intralayer bonding. This work exemplifies the evolution of structural parameters in layered materials in changing from the 3-dimensional to the 2-dimensional regime.Comment: 14 pages, 4 figure