Mice Lacking Gpr37 Exhibit Decreased Expression of the Myelin-Associated Glycoprotein Mag and Increased Susceptibility to Demyelination

GPR37 is an orphan G protein-coupled receptor that is predominantly expressed in the brain and found at particularly high levels in oligodendrocytes. GPR37 has been shown to exert effects on oligodendrocyte differentiation and myelination during development, but the molecular basis of these actions is incompletely understood and moreover nothing is known about the potential role(s) of this receptor under demyelinating conditions. To shed light on the fundamental biology of GPR37, we performed proteomic studies comparing protein expression levels in the brains of mice lacking GPR37 and its close relative GPR37-like 1 (GPR37L1). These studies revealed that one of the proteins most sharply decreased in the brains of Gpr37/Gpr37L1 double knockout mice is the myelin-associated glycoprotein MAG. Follow-up Western blot studies confirmed this finding and demonstrated that genetic deletion of Gpr37, but not Gpr37L1, results in strikingly decreased brain expression of MAG. Further in vitro studies demonstrated that GPR37 and MAG form a complex when expressed together in cells. As loss of MAG has previously been shown to result in increased susceptibility to brain insults, we additionally assessed Gpr37-knockout (Gpr37−/−) vs. wild-type mice in the cuprizone model of demyelination. These studies revealed that Gpr37−/− mice exhibit dramatically increased loss of myelin in response to cuprizone, yet do not show any increased loss of oligodendrocyte precursor cells or mature oligodendrocytes. These findings reveal that loss of GPR37 alters oligodendrocyte physiology and increases susceptibility to demyelination, indicating that GPR37 could be a potential drug target for the treatment of demyelinating diseases such as multiple sclerosis

A New Human NHERF1 Mutation Decreases Renal Phosphate Transporter NPT2a Expression by a PTH-Independent Mechanism

Background: The sodium-hydrogen exchanger regulatory factor 1 (NHERF1) binds to the main renal phosphate transporter NPT2a and to the parathyroid hormone (PTH) receptor. We have recently identified mutations in NHERF1 that decrease renal phosphate reabsorption by increasing PTH-induced cAMP production in the renal proximal tubule. Methods: We compared relevant parameters of phosphate homeostasis in a patient with a previously undescribed mutation in NHERF1 and in control subjects. We expressed the mutant NHERF1 protein in Xenopus Oocytes and in cultured cells to study its effects on phosphate transport and PTH-induced cAMP production. Results: We identified in a patient with inappropriate renal phosphate reabsorption a previously unidentified mutation (E68A) located in the PDZ1 domain of NHERF1.We report the consequences of this mutation on NHERF1 function. E68A mutation did not modify cAMP production in the patient. PTH-induced cAMP synthesis and PKC activity were not altered by E68A mutation in renal cells in culture. In contrast to wild-type NHERF1, expression of the E68A mutant in Xenopus oocytes and in human cells failed to increase phosphate transport. Pull down experiments showed that E68A mutant did not interact with NPT2a, which robustly interacted with wild type NHERF1 and previously identified mutants. Biotinylation studies revealed that E68A mutant was unable to increase cell surface expression of NPT2a. Conclusions: Our results indicate that the PDZ1 domain is critical for NHERF1- NPT2a interaction in humans and for th

User's Guide Virtual Hydropower Prospector Version 1.1

The Virtual Hydropower Prospector is a web-based geographic information system (GIS) application for displaying U.S. water energy resource sites on hydrologic region maps. The application assists the user in locating sites of interest and performing preliminary, development feasibility assessments. These assessments are facilitated by displaying contextual features in addition to the water energy resource sites such as hydrograpy, roads, power infrastructure, populated places, and land use and control. This guide provides instructions for operating the application to select what features are displayed and the extent of the map view. It also provides tools for selecting features of particular interest and displaying their attribute information

Quantitative Proteomics Reveal an Altered Pattern of Protein Expression in Brain Tissue from Mice Lacking Gpr37 and Gpr37l1

GPR37 and GPR37L1 are glia-enriched G protein-coupled receptors that have been implicated in several neurological and neurodegenerative diseases. To gain insight into the potential molecular mechanisms by which GPR37 and GPR37L1 regulate cellular physiology, proteomic analyses of whole mouse brain tissue from wild-type (WT) versus GPR37/GPR37L1 double knockout (DKO) mice were performed in order to identify proteins regulated by the absence versus presence of these receptors (data are available via ProteomeXchange with identifier PXD015202). These analyses revealed a number of proteins that were significantly increased or decreased by the absence of GPR37 and GPR37L1. One of the most decreased proteins in the DKO versus WT brain tissue was S100A5, a calcium-binding protein, and the reduction of S100A5 expression in KO brain tissue was validated via Western blot. Coexpression of S100A5 with either GPR37 or GPR37L1 in HEK293T cells did not result in any change in S100A5 expression but did robustly increase secretion of S100A5. To dissect the mechanism by which S100A5 secretion was enhanced, cells coexpressing S100A5 with the receptors were treated with different pharmacological reagents. These studies revealed that calcium is essential for the secretion of S100A5 downstream of GPR37 and GPR37L1 signaling, as treatment with BAPTA-AM, an intracellular Ca2+ chelator, reduced S100A5 secretion from transfected HEK293T cells. Collectively, these findings provide a panoramic view of proteomic changes resulting from loss of GPR37 and GPR37L1 and also impart mechanistic insight into the regulation of S100A5 by these receptors, thereby shedding light on the functions of GPR37 and GPR37L1 in brain tissue

Extracellular Calcium Modulates Actions of Orthosteric and Allosteric Ligands on Metabotropic Glutamate Receptor 1alpha

SUMMARY: Metabotropic glutamate receptor 1α (mGluR1α), a member of the family C G protein-coupled receptors (GPCRs), is emerging as a potential drug target for various disorders including chronic neuronal degenerative diseases. In addition to being activated by glutamate, mGluR1α is also modulated by extracellular Ca2+. However, the underlying mechanism is unknown. Moreover, it has long been challenging to develop receptor-specific agonists due to homologies within the mGluR family, and the Ca2+-binding site(s) on mGluR1α may provide an opportunity for receptor-selective targeting by therapeutics. In the present study, we show that our previously predicted Ca2+-binding site in the hinge region of mGluR1α is adjacent to the site where orthosteric agonists and antagonists bind on the extracellular domain of the receptor. Moreover, we have found that extracellular Ca2+ enhances mGluR1α-mediated intracellular Ca2+ responses evoked by the orthosteric agonist, L-quisqualate. Conversely, extracellular Ca2+ diminishes the inhibitory effect of the mGluR1α orthosteric antagonist, (s)-MCPG. In addition, selective positive (Ro 67-4853) and negative (CPCCOEt) allosteric modulators of mGluR1α potentiate and inhibit responses to extracellular Ca2+, respectively, in a manner similar to their effects on the response of mGluR1α to glutamate. Mutations at residues predicted to be involved in Ca2+-binding, including E325I, have significant effects on the modulation of responses to the orthosteric agonist, L-quisqualate, and the allosteric modulator Ro 67-4853 by extracellular Ca2+. These studies reveal that binding of extracellular Ca2+ to the predicted Ca2+-binding site in the ECD of mGluR1α modulates not only glutamate-evoked signaling but also the actions of both orthosteric ligands and allosteric modulators on mGluR1α

Star formation in 30 Doradus

Using observations obtained with the Wide Field Camera 3 (WFC3) on board the Hubble Space Telescope (HST), we have studied the properties of the stellar populations in the central regions of 30 Dor, in the Large Magellanic Cloud. The observations clearly reveal the presence of considerable differential extinction across the field. We characterise and quantify this effect using young massive main sequence stars to derive a statistical reddening correction for most objects in the field. We then search for pre-main sequence (PMS) stars by looking for objects with a strong (> 4 sigma) Halpha excess emission and find about 1150 of them over the entire field. Comparison of their location in the Hertzsprung-Russell diagram with theoretical PMS evolutionary tracks for the appropriate metallicity reveals that about one third of these objects are younger than ~4Myr, compatible with the age of the massive stars in the central ionising cluster R136, whereas the rest have ages up to ~30Myr, with a median age of ~12Myr. This indicates that star formation has proceeded over an extended period of time, although we cannot discriminate between an extended episode and a series of short and frequent bursts that are not resolved in time. While the younger PMS population preferentially occupies the central regions of the cluster, older PMS objects are more uniformly distributed across the field and are remarkably few at the very centre of the cluster. We attribute this latter effect to photoevaporation of the older circumstellar discs caused by the massive ionising members of R136.Comment: 15 pages, 12 figures. Accepted for publication in The Astrophysical Journa

The Luminosity, Mass, and Age Distributions of Compact Star Clusters in M83 Based on HST/WFC3 Observations

The newly installed Wide Field Camera 3 (WFC3) on the Hubble Space Telescope has been used to obtain multi-band images of the nearby spiral galaxy M83. These new observations are the deepest and highest resolution images ever taken of a grand-design spiral, particularly in the near ultraviolet, and allow us to better differentiate compact star clusters from individual stars and to measure the luminosities of even faint clusters in the U band. We find that the luminosity function for clusters outside of the very crowded starburst nucleus can be approximated by a power law, dN/dL \propto L^{alpha}, with alpha = -2.04 +/- 0.08, down to M_V ~ -5.5. We test the sensitivity of the luminosity function to different selection techniques, filters, binning, and aperture correction determinations, and find that none of these contribute significantly to uncertainties in alpha. We estimate ages and masses for the clusters by comparing their measured UBVI,Halpha colors with predictions from single stellar population models. The age distribution of the clusters can be approximated by a power-law, dN/dt propto t^{gamma}, with gamma=-0.9 +/- 0.2, for M > few x 10^3 Msun and t < 4x10^8 yr. This indicates that clusters are disrupted quickly, with ~80-90% disrupted each decade in age over this time. The mass function of clusters over the same M-t range is a power law, dN/dM propto M^{beta}, with beta=-1.94 +/- 0.16, and does not have bends or show curvature at either high or low masses. Therefore, we do not find evidence for a physical upper mass limit, M_C, or for the earlier disruption of lower mass clusters when compared with higher mass clusters, i.e. mass-dependent disruption. We briefly discuss these implications for the formation and disruption of the clusters.Comment: 36 pages, 13 figures, 1 table; accepted for publication in the Astrophysical Journa