Segregation of Fluorescent Membrane Lipids into Distinct Micrometric Domains: Evidence for Phase Compartmentation of Natural Lipids?

Background: We recently reported that sphingomyelin (SM) analogs substituted on the alkyl chain by various fluorophores (e.g. BODIPY) readily inserted at trace levels into the plasma membrane of living erythrocytes or CHO cells and spontaneously concentrated into micrometric domains. Despite sharing the same fluorescent ceramide backbone, BODIPY-SM domains segregated from similar domains labelled by BODIPY-D-e-lactosylceramide (D-e-LacCer) and depended on endogenous SM. Methodology/Principal Findings. We show here that BODIPY-SM further differed from BODIPY-D-e-LacCer or -glucosylceramide (GlcCer) domains in temperature dependence, propensity to excimer formation, association with a glycosylphosphatidylinositol (GPI)-anchored fluorescent protein reporter, and lateral diffusion by FRAP, thus demonstrating different lipid phases and boundaries. Whereas BODIPY-D-e-LacCer behaved like BODIPY-GlcCer, its artificial stereoisomer, BODIPY-L-t-LacCer, behaved like BODIPY- and NBD-phosphatidylcholine (PC). Surprisingly, these two PC analogs also formed micrometric patches yet preferably at low temperature, did not show excimer, never associated with the GPI reporter and showed major restriction to lateral diffusion when photobleached in large fields. This functional comparison supported a three-phase micrometric compartmentation, of decreasing order: BODIPY-GSLs > -SM > -PC (or artificial L-t-LacCer). Co-existence of three segregated compartments was further supported by double labelling experiments and was confirmed by additive occupancy, up to ~70% cell surface coverage. Specific alterations of BODIPY-analogs domains by manipulation of corresponding endogenous sphingolipids suggested that distinct fluorescent lipid partition might reflect differential intrinsic propensity of endogenous membrane lipids to form large assemblies. Conclusions/Significance. We conclude that fluorescent membrane lipids spontaneously concentrate into distinct micrometric assemblies. We hypothesize that these might reflect preexisting compartmentation of endogenous PM lipids into non-overlapping domains of differential order: GSLs > SM > PC, resulting into differential self-adhesion of the two former, with exclusion of the latter

Oracle Efficient Estimation of Structural Breaks in Cointegrating Regressions

In this paper, we propose an adaptive group lasso procedure to efficiently estimate structural breaks in cointegrating regressions. It is well-known that the group lasso estimator is not simultaneously estimation consistent and model selection consistent in structural break settings. Hence, we use a first step group lasso estimation of a diverging number of breakpoint candidates to produce weights for a second adaptive group lasso estimation. We prove that parameter changes are estimated consistently by group lasso and show that the number of estimated breaks is greater than the true number but still sufficiently close to it. Then, we use these results and prove that the adaptive group lasso has oracle properties if weights are obtained from our first step estimation. Simulation results show that the proposed estimator delivers the expected results. An economic application to the long-run US money demand function demonstrates the practical importance of this methodology

Phosphorylation of SCG10/stathmin-2 determines multipolar stage exit and neuronal migration rate

Cristina García-Frigola [et al.]. 11 p., 7 figures, 1 table and references.Cell migration is the consequence of the sum of positive and negative regulatory mechanisms. Although appropriate migration of neurons is a principal feature of brain development, the negative regulatory mechanisms remain obscure. We found that JNK1 was highly active in developing cortex and that selective inhibition of JNK in the cytoplasm markedly increased both the frequency of exit from the multipolar stage and radial migration rate and ultimately led to an ill-defined cellular organization. Moreover, regulation of multipolar-stage exit and radial migration in Jnk1 -/- (also known as Mapk8) mice, resulted from consequential changes in phosphorylation of the microtubule regulator SCG10 (also called stathmin-2). Expression of an SCG10 mutant that mimics the JNK1-phosphorylated form restored normal migration in the brains of Jnk1 -/- mouse embryos. These findings indicate that the phosphorylation of SCG10 by JNK1 is a fundamental mechanism that governs the transition from the multipolar stage and the rate of neuronal cell movement during cortical development.This work was supported by grants from the Academy of Finland (218125, 125860, 206497, 111870, 203520 and 110445), Turku Graduate School of Biomedical Sciences, the Finnish Graduate School of Neuroscience, Magnus Ehrnrooth’s Foundation, FP6 STRESSPROTECT and Åbo Akademi University. T.K. was supported by the Danish Cancer Society. L.N. is funded by Walloon Excellence in Lifesciences and Biotechnology.Peer reviewe