Antisense-mediated isoform switching of steroid receptor coactivator-1 in the central nucleus of the amygdala of the mouse brain

Background

Antisense oligonucleotide (AON)-mediated exon skipping is a powerful tool to manipulate gene expression. In the present study we investigated the potential of exon skipping by local injection in the central nucleus of the amygdala (CeA) of the mouse brain. As proof of principle we targeted the splicing of steroid receptor coactivator-1 (SRC-1), a protein involved in nuclear receptor function. This nuclear receptor coregulator exists in two splice variants (SRC-1a and SRC-1e) which display differential distribution and opposing activities in the brain, and whose mRNAs differ in a single SRC-1e specific exon.

Methods

For proof of principle of feasibility, we used immunofluorescent stainings to study uptake by different cell types, translocation to the nucleus and potential immunostimulatory effects at different time points after a local injection in the CeA of the mouse brain of a control AON targeting human dystrophin with no targets in the murine brain. To evaluate efficacy we designed an AON targeting the SRC-1e-specific exon and with qPCR analysis we measured the expression ratio of the two splice variants.

Results

We found that AONs were taken up by corticotropin releasing hormone expressing neurons and other cells in the CeA, and translocated into the cell nucleus. Immune responses after AON injection were comparable to those after sterile saline injection. A successful shift of the naturally occurring SRC-1a:SRC-1e expression ratio in favor of SRC-1a was observed, without changes in total SRC-1 expression.

Conclusions

We provide a proof of concept for local neuropharmacological use of exon skipping by manipulating the expression ratio of the two splice variants of SRC-1, which may be used to study nuclear receptor function in specific brain circuits. We established that exon skipping after local injection in the brain is a versatile and useful tool for the manipulation of splice variants for numerous genes that are relevant for brain function.     

The Lyapunov Exponent of Products of Random 2×2 Matrices Close to the Identity

We study products of arbitrary random real 2×2 matrices that are close to the identity matrix. Using the Iwasawa decomposition of SL(2,?), we identify a continuum regime where the mean values and the covariances of the three Iwasawa parameters are simultaneously small. In this regime, the Lyapunov exponent of the product is shown to assume a scaling form. In the general case, the corresponding scaling function is expressed in terms of Gauss’ hypergeometric function. A number of particular cases are also considered, where the scaling function of the Lyapunov exponent involves other special functions (Airy, Bessel, Whittaker, elliptic). The general solution thus obtained allows us, among other things, to recover in a unified framework many results known previously from exactly solvable models of one-dimensional disordered systems.     

Air-dispersion measurement by second-harmonic heterodyne interferometry

Interferometers with long optical paths in air usually require knowledge and control of air dispersion. In addition, the measurements at several wavelengths and the dispersion properties of air allow errors caused by air turbulence to be compensated for. An innovative technique for air-dispersion measurement is described for long-baseline ground-based stellar interferometers. The technique combines second-harmonic interferometry and heterodyne detection to permit high-resolution measurement even for low optical powers. Experimental results show measurements of air dispersion in good agreement with the values predicted from the Edlén equation.     

Probing Proteins in Solution by Xe NMR Spectroscopy

The interaction of xenon with different proteins in aqueous solution is investigated by ¹²⁹Xe NMR spectroscopy. Chemical shifts are measured in horse metmyoglobin, hen egg white lysozyme, and horse cytochrome c solutions as a function of xenon concentration. In these systems, xenon is in fast exchange between all possible environments. The results suggest that nonspecific interactions exist between xenon and the protein exteriors and the data are analyzed in term of parameters which characterize the protein surfaces. The experimental data for horse metmyoglobin are interpreted using a model in which xenon forms a 1:1 complex with the protein and the chemical shift of the complexed xenon is reported (Locci et al., Keystone Symposia “Frontiers of NMR in Molecular Biology VI”, Jan. 9–15, 1999, Breckenridge, CO, Abstract E216, p. 53; Locci et al., XeMAT 2000 “Optical Polarization and Xenon NMR of Materials”, June 28–30, 2000, Sestri Levante, Italy, p. 46).     

An original approach for Raman spectroscopy analysis of radioactive materials and its application to americium‐containing samples

An approach for Raman measurements of highly radioactive samples is presented here. The innovative part of this approach lies in the fact that no single part of the Raman equipment is in direct contact with the radioactive sample, as the sample is sealed in an alpha‐tight capsule. Raman analysis is effectively performed through the optical‐grade quartz window closing the capsule. This allows performing micro‐Raman measurements on radioactive samples with no limitations on the laser source wavelength, polarisation mode, spectrometer mode and microscope mode (provided the focal length of the microscope objective is greater than the thickness of the quartz window and with sub mg samples). Some example results are shown and discussed. In particular, some spectral features of americium‐containing oxide nuclear fuel specimens are presented. Raman spectra clearly reveal in these specimens the presence of abundant oxygen defects induced in the fcc fluorite lattice by trivalent americium. In order to complete the analysis the Raman spectrum of pure americium dioxide was also measured with a lower energy excitation source compared with previous research. The current results seem to be consistent with the possible occurrence of a photolysis process induced by the Raman laser, resulting in the formation of hyperstoichiometric americium sesquioxide Am₂O_{3 + z}. Such a photolytic process is deemed to be unavoidable when visible lasers are used as excitation sources for the Raman analysis of americium dioxide. © 2015 The Authors Journal of Raman Spectroscopy Published by John Wiley & Sons, Ltd.     

Role of nitrogen in the mobility drop of electrons in modulation-doped GaAsN/AlGaAs heterostructures

We report transport properties of a 2 dimension electron gas (2DEG) in molecular beam epitaxy-grown GaAs_1−xN_x/AlGaAs modulation-doped heterostructures. Quantum oscillations in far infrared cyclotron resonance prove the efficient electron transfer and formation of the 2DEG. The 2DEG mobility strongly depends on the N concentration in the channel layer. It shows a drastic decrease as compared to N-free samples, even for the smallest amount of N (0.02%). For this N composition, the electron effective mass was found to be 0.073m₀. Reduced growth temperature (450 °C) was found to improve the mobility of N-containing channels. Examination of transport properties from 4 to 300 K and cyclotron resonance experiments give evidence of the presence of ionised impurity-like scattering centres in GaAsN.     

Failure of the Ingard-Myers boundary condition for a lined duct: an experimental investigation

This paper deals with experimental investigation of the lined wall boundary condition in flow duct applications such as aircraft engine systems or automobile mufflers. A first experiment, based on a microphone array located in the liner test section, is carried out in order to extract the axial wavenumbers with the help of an "high-accurate" singular value decomposition Prony-like algorithm. The experimental axial wavenumbers are then used to provide the lined wall impedance for both downstream and upstream acoustic propagation by means of a straightforward impedance education method involving the classical Ingard-Myers boundary condition. The results show that the Ingard-Myers boundary condition fails to predict with accuracy the acoustic behavior in a lined duct with flow. An effective lined wall impedance, valid whatever the direction of acoustic propagation, can be suitably found from experimental axial wavenumbers and a modified version of the Ingard-Myers condition with the form inspired from a previous theoretical study [Aure?gan et al., J. Acoust. Soc. Am. 109, 59-64 (2001)]. In a second experiment, the scattering matrix of the liner test section is measured and is then compared to the predicted scattering matrix using the multimodal approach and the lined wall impedances previously deduced. A large discrepancy is observed between the measured and the predicted scattering coefficients that confirms the poor accuracy provided from the Ingard-Myers boundary condition widely used in lined duct applications.     

Entropy-stabilized smectic C phase in a system of zigzag-shaped molecules

We report Monte Carlo simulations of a system of rigid zigzag-shaped molecules that demonstrate that simple excluded-volume interactions are sufficient to produce a fluid tilted lamellar [smectic C (SmC)] liquid crystal phase. The molecules are composed of three rigidly linked hard spherocylinders arranged in a zigzag fashion. By varying the zigzag angle we have mapped out the whole phase diagram as a function of pressure and zigzag angle Psi. For Psi between 35 degrees and 80 degrees our model simulation exhibits the SmC phase. This is the first conclusive evidence where steric interactions arising out of molecular shape alone induce the occurrence of the SmC phase for a wide range of zigzag angles. For smaller Psi, a transition from tilted crystal to crystal is observed.