Sortilin, SorCS1b, and SorLA Vps10p sorting receptors, are novel γ-secretase substrates

BACKGROUND: The mammalian Vps10p sorting receptor family is a group of 5 type I membrane homologs (Sortilin, SorLA, and SorCS1-3). These receptors bind various cargo proteins via their luminal Vps10p domains and have been shown to mediate a variety of intracellular sorting and trafficking functions. These proteins are highly expressed in the brain. SorLA has been shown to be down regulated in Alzheimer's disease brains, interact with ApoE, and modulate Aβ production. Sortilin has been shown to be part of proNGF mediated death signaling that results from a complex of Sortilin, p75(NTR )and proNGF. We have investigated and provide evidence for γ-secretase cleavage of this family of proteins. RESULTS: We provide evidence that these receptors are substrates for presenilin dependent γ-secretase cleavage. γ-Secretase cleavage of these sorting receptors is inhibited by γ-secretase inhibitors and does not occur in PS1/PS2 knockout cells. Like most γ-secretase substrates, we find that ectodomain shedding precedes γ-secretase cleavage. The ectodomain cleavage is inhibited by a metalloprotease inhibitor and activated by PMA suggesting that it is mediated by an α-secretase like cleavage. CONCLUSION: These data indicate that the α- and γ-secretase cleavages of the mammalian Vps10p sorting receptors occur in a fashion analogous to other known γ-secretase substrates, and could possibly regulate the biological functions of these proteins

Signal peptide peptidase (SPP) dimer formation as assessed by fluorescence lifetime imaging microscopy (FLIM) in intact cells

BACKGROUND: Signal peptide peptidase (SPP) is an intramembrane cleaving protease identified by its cleavage of several type II membrane signal peptides. Conservation of intramembrane active site residues demonstrates that SPP, SPP family members, and presenilins (PSs) make up a family of intramembrane cleaving proteases. Because SPP appears to function without additional protein cofactors, the study of SPP may provide structural insights into the mechanism of intramembrane proteolysis by this biomedically important family of proteins. Previous studies have shown that SPP isolated from cells appears to be a homodimer, but some evidence exists that in vitro SPP may be active as a monomer. We have conducted additional experiments to determine if SPP exists as a monomer or dimer in vivo. RESULTS: Fluorescence lifetime imaging microscopy (FLIM) can be is used to determine intra- or intermolecular interactions by fluorescently labeling epitopes on one or two different molecules. If the donor and acceptor fluorophores are less than 10 nm apart, the donor fluorophore lifetime shortens proportionally to the distance between the fluorophores. In this study, we used two types of fluorescence energy transfer (FRET) pairs; cyan fluorescent protein (CFP) with yellow fluorescent protein (YFP) or Alexa 488 with Cy3 to differentially label the NH2- or COOH-termini of SPP molecules. A cell based SPP activity assay was used to show that all tagged SPP proteins are proteolytically active. Using FLIM we were able to show that the donor fluorophore lifetime of the CFP tagged SPP construct in living cells significantly decreases when either a NH2- or COOH-terminally YFP tagged SPP construct is co-transfected, indicating close proximity between two different SPP molecules. These data were then confirmed in cell lines stably co-expressing V5- and FLAG-tagged SPP constructs. CONCLUSION: Our FLIM data strongly suggest dimer formation between two separate SPP proteins. Although the tagged SPP constructs are expressed throughout the cell, SPP dimer detection by FLIM is seen predominantly at or near the plasma membrane

Cholestenoic acid, an endogenous cholesterol metabolite, is a potent γ-secretase modulator.

BackgroundAmyloid-β (Aβ) 42 has been implicated as the initiating molecule in the pathogenesis of Alzheimer's disease (AD); thus, therapeutic strategies that target Aβ42 are of great interest. γ-Secretase modulators (GSMs) are small molecules that selectively decrease Aβ42. We have previously reported that many acidic steroids are GSMs with potencies ranging in the low to mid micromolar concentration with 5β-cholanic acid being the most potent steroid identified GSM with half maximal effective concentration (EC50) of 5.7 μM.ResultsWe find that the endogenous cholesterol metabolite, 3β-hydroxy-5-cholestenoic acid (CA), is a steroid GSM with enhanced potency (EC50 of 250 nM) relative to 5β-cholanic acid. CA i) is found in human plasma at ~100-300 nM concentrations ii) has the typical acidic GSM signature of decreasing Aβ42 and increasing Aβ38 levels iii) is active in in vitro γ-secretase assay iv) is made in the brain. To test if CA acts as an endogenous GSM, we used Cyp27a1 knockout (Cyp27a1-/-) and Cyp7b1 knockout (Cyp7b1-/-) mice to investigate if manipulation of cholesterol metabolism pathways relevant to CA formation would affect brain Aβ42 levels. Our data show that Cyp27a1-/- had increased brain Aβ42, whereas Cyp7b1-/- mice had decreased brain Aβ42 levels; however, peripheral dosing of up to 100 mg/kg CA did not affect brain Aβ levels. Structure-activity relationship (SAR) studies with multiple known and novel CA analogs studies failed to reveal CA analogs with increased potency.ConclusionThese data suggest that CA may act as an endogenous GSM within the brain. Although it is conceptually attractive to try and increase the levels of CA in the brain for prevention of AD, our data suggest that this will not be easily accomplished

Molecular line mapping of the giant molecular cloud associated with RCW 106 - II. Column density and dynamical state of the clumps

We present a fully sampled C^{18}O (1-0) map towards the southern giant molecular cloud (GMC) associated with the HII region RCW 106, and use it in combination with previous ^{13}CO (1-0) mapping to estimate the gas column density as a function of position and velocity. We find localized regions of significant ^{13}CO optical depth in the northern part of the cloud, with several of the high-opacity clouds in this region likely associated with a limb-brightened shell around the HII region G333.6-0.2. Optical depth corrections broaden the distribution of column densities in the cloud, yielding a log-normal distribution as predicted by simulations of turbulence. Decomposing the ^{13}CO and C^{18}O data cubes into clumps, we find relatively weak correlations between size and linewidth, and a more sensitive dependence of luminosity on size than would be predicted by a constant average column density. The clump mass spectrum has a slope near -1.7, consistent with previous studies. The most massive clumps appear to have gravitational binding energies well in excess of virial equilibrium; we discuss possible explanations, which include magnetic support and neglect of time-varying surface terms in the virial theorem. Unlike molecular clouds as a whole, the clumps within the RCW 106 GMC, while elongated, appear to show random orientations with respect to the Galactic plane.Comment: 17 pages, to appear in MNRA

Researcher Degrees of Freedom in the Psychology of Religion

There is a push in psychology toward more transparent practices, stemming partially as a response to the replication crisis. We argue that the psychology of religion should help lead the way toward these new, more transparent practices to ensure a robust and dynamic subfield. One of the major issues that proponents of Open Science practices hope to address is researcher degrees of freedom (RDF). We pre-registered and conducted a systematic review of the 2017 issues from three psychology of religion journals. We aimed to identify the extent to which the psychology of religion has embraced Open Science practices and the role of RDF within the subfield. We found that many of the methodologies that help to increase transparency, such as pre-registration, have yet to be adopted by those in the subfield. In light of these findings, we present recommendations for addressing the issue of transparency in the psychology of religion and outline how to move toward these new Open Science practices

Habitat quality differentiation and consequences for ecosystem service provision of an amazonian hyperdominant tree species.

Ecosystem services of Amazonian forests are disproportionally produced by a limited set of hyperdominant tree species. Yet the spatial variation in the delivery of ecosystem services by individual hyperdominant species across their distribution ranges and corresponding environmental gradients is poorly understood. Here, we use the concept of habitat quality to unravel the effect of environmental gradients on seed production and aboveground biomass (AGB) of the Brazil nut, one of Amazonia's largest and most long-lived hyperdominants. We find that a range of climate and soil gradients create trade-offs between density and fitness of Brazil nut trees. Density responses to environmental gradients were in line with predictions under the Janzen-Connell and Herms-Mattson hypotheses, whereas tree fitness responses were in line with resource requirements of trees over their life cycle. These trade-offs resulted in divergent responses in area-based seed production and AGB. While seed production and AGB of individual trees (i.e., fitness) responded similarly to most environmental gradients, they showed opposite tendencies to tree density for almost half of the gradients. However, for gradients creating opposite fitness-density responses, area-based seed production was invariable, while trends in area-based AGB tended to mirror the response of tree density. We conclude that while the relation between environmental gradients and tree density is generally indicative of the response of AGB accumulation in a given area of forest, this is not necessarily the case for fruit production

Global regulation of alternative splicing during myogenic differentiation

Recent genome-wide analyses have elucidated the extent of alternative splicing (AS) in mammals, often focusing on comparisons of splice isoforms between differentiated tissues. However, regulated splicing changes are likely to be important in biological transitions such as cellular differentiation, or response to environmental stimuli. To assess the extent and significance of AS in myogenesis, we used splicing-sensitive microarray analysis of differentiating C2C12 myoblasts. We identified 95 AS events that undergo robust splicing transitions during C2C12 differentiation. More than half of the splicing transitions are conserved during differentiation of avian myoblasts, suggesting the products and timing of transitions are functionally significant. The majority of splicing transitions during C2C12 differentiation fall into four temporal patterns and were dependent on the myogenic program, suggesting that they are integral components of myogenic differentiation. Computational analyses revealed enrichment of many sequence motifs within the upstream and downstream intronic regions near the alternatively spliced regions corresponding to binding sites of splicing regulators. Western analyses demonstrated that several splicing regulators undergo dynamic changes in nuclear abundance during differentiation. These findings show that within a developmental context, AS is a highly regulated and conserved process, suggesting a major role for AS regulation in myogenic differentiation.National Institutes of Health (U.S.) (grant number R01GM076493)Ford Foundation (Predoctoral Diversity Fellowship)Baylor College of Medicine. Graduate School of Biomedical Sciences (Baylor Research Advocates for Student Scientists

Psychology as a natural science in the eighteenth century

Psychology considered as a natural science began as Aristotelian "physics" or "natural philosophy" of the soul. C. Wolff placed psychology under metaphysics, coordinate with cosmology. Scottish thinkers placed it within moral philosophy, but distinguished its "physical" laws from properly moral laws (for guiding conduct). Several Germans sought to establish an autonomous empirical psychology as a branch of natural science. British and French visual theorists developed mathematically precise theories of size and distance perception; they created instruments to test these theories and to measure visual phenomena such as the duration of visual impressions. These investigators typically were dualists who included mental phenomena within nature