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.     

Late-Stage Functionalization by Chan–Lam Amination: Rapid Access to Potent and Selective Integrin Inhibitors

A late-stage functionalization of the aromatic ring in amino acid derivatives is described. The key step is a copper-catalysed diversification of a boronate ester by amination (Chan–Lam reaction) that can be carried out on a complex β-aryl-β-amino acid scaffold. This not only considerably extends the substrate scope of amination partners, but also delivers an array of potent and selective integrin inhibitors as potential treatment agents of idiopathic pulmonary fibrosis (IPF). This versatile chemical strategy, which is amenable to high-throughput-array protocols, allows the installation of pharmaceutically valuable heteroaromatic fragments at a late stage by direct coupling to NH heterocycles, leading to compounds with drug-like attributes. It thus constitutes a useful addition to the medicinal chemist's repertoire.     

The survival probability for critical spread-out oriented percolation above 4 + 1 dimensions. I. Induction

We consider critical spread-out oriented percolation above 4 + 1 dimensions. Our main result is that the extinction probability at time n (i.e., the probability for the origin to be connected to the hyperplane at time n but not to the hyperplane at time n + 1) decays like 1/Bn ² as , where B is a finite positive constant. This in turn implies that the survival probability at time n (i.e., the probability that the origin is connected to the hyperplane at time n) decays like 1/Bn as . The latter has been shown in an earlier paper to have consequences for the geometry of large critical clusters and for the incipient infinite cluster. The present paper is Part I in a series of two papers. In Part II, we derive a lace expansion for the survival probability, adapted so as to deal with point-to-plane connections. This lace expansion leads to a nonlinear recursion relation for the survival probability. In Part I, we use this recursion relation to deduce the asymptotics via induction.     

Novel software for the assignment of peaks from tandem mass spectrometry spectra of synthetic polymers

Novel software has been developed to aid the interpretation of tandem mass spectrometry (MS/MS) data from synthetic polymers. The software is particularly focused toward aiding the end-group determination of these materials by significantly speeding up the interpretation process. This allows information on the initiator and/or chain transfer agents, used to generate the polymer, and the mechanism of termination to be inferred from the data much more rapidly. The software allows the validity of hypothesized structures to be rapidly tested by automatically annotating the data file using previously proposed fragmentation rules for synthetic polymers. Low-energy collision-induced dissociation (CID) data from methacrylate, styrene, and polyether oligomers are used as example data for the software. Exact-mass CID information was used to aid the understanding of the dissociation mechanism of the polymers. The software can use exact-mass data to provide more confidence in the results. The MS/MS results indicate that the fragmentation pathways are those previously proposed for these polymers.     

Transfer measurements for the Ti+Ni systems at near barrier energies

Large enhancements have been observed in the sub-barrier fusion cross sections for Ti+Ni systems in our previous studies. Coupled channel calculations incorporating couplings to 2⁺ and 3⁻ states failed to explain these enhancements completely. A possibilty of transfer channels contributing to the residual enhancements had been suggested. In order to investigate the role of relevant transfer channels, measurements of one- and two-nucleon transfer were carried out for ^46,48Ti+⁶¹Ni systems. The present paper gives the results of these studies.     

Carbon papers with conductive polymer coatings for use in solid-state supercapacitors

Conductive polymer-coated carbon papers have been fabricated through polymerisation of pyrrole-based monomers oxidised with various heteropolyacids. Smooth surfaces are obtained when multiple coatings are applied to the carbon surface and give good contact with the Nafion^® electrolyte. Cyclic voltammetry was used to study the electrodes and a.c. impedance and charge / discharge cycling was used to study membrane electrode assemblies (MEA). MEAs were fabricated using a hot-press technique.     

Scaling Limits and Critical Behaviour of the 4-Dimensional n-Component |\varphi |^4 Spin Model

We consider the \(n\) -component \(|\varphi |^4\) spin model on \({\mathbb {Z}}^4\) , for all \(n \ge 1\) , with small coupling constant. We prove that the susceptibility has a logarithmic correction to mean field scaling, with exponent \(\frac{n+2}{n+8}\) for the logarithm. We also analyse the asymptotic behaviour of the pressure as the critical point is approached, and prove that the specific heat has fractional logarithmic scaling for \(n =1,2,3\) ; double logarithmic scaling for \(n=4\) ; and is bounded when \(n>4\) . In addition, for the model defined on the \(4\) -dimensional discrete torus, we prove that the scaling limit as the critical point is approached is a multiple of a Gaussian free field on the continuum torus, whereas, in the subcritical regime, the scaling limit is Gaussian white noise with intensity given by the susceptibility. The proofs are based on a rigorous renormalisation group method in the spirit of Wilson, developed in a companion series of papers to study the 4-dimensional weakly self-avoiding walk, and adapted here to the \(|\varphi |^4\) model.     

The Scaling Limit of Lattice Trees in High Dimensions

We prove that above eight dimensions the scaling limit of sufficiently spread-out lattice trees is the variant of super-Brownian motion known as integrated super-Brownian excursion (ISE), as conjectured by Aldous. The same is true for nearest-neighbour lattice trees in sufficiently high dimensions. The proof uses the lace expansion. Received: 27 September 1996 / Accepted: 28 July 1997