MANF regulates neuronal survival and UPR through its ER-located receptor IRE1a

Mesencephalic astrocyte-derived neurotrophic factor (MANF) is an endoplasmic reticulum (ER)-located pro-tein with cytoprotective effects in neurons and pancreatic b cells in vitro and in models of neurodegeneration and diabetes in vivo. However, the exact mode of MANF action has remained elusive. Here, we show that MANF directly interacts with the ER transmembrane unfolded protein response (UPR) sensor IRE1a, and we identify the binding interface between MANF and IRE1a. The expression of wild-type MANF, but not its IRE1a binding-deficient mutant, attenuates UPR signaling by decreasing IRE1a oligomerization; phosphor-ylation; splicing of Xbp1, Atf6, and Txnip levels; and protecting neurons from ER stress-induced death. MANF-IRE1a interaction and not MANF-BiP interaction is crucial for MANF pro-survival activity in neurons in vitro and is required to protect dopamine neurons in an animal model of Parkinson's disease. Our data show IRE1a as an intracellular receptor for MANF and regulator of neuronal survival.Peer reviewe

Forecasts of the Scottish economy [February 2009]

The global economy is experiencing a significant contraction due to the spreading effects of the sub-prime and financial crisis. This has greater effects in the main developed economies (US, UK, France, Germany and Japan) but the repercussions affect countries such as Russia, China and India, where a sharp downturn in global growth adversely impacts on their growth

Telerobotic research at NASA Langley Research Center

An overview of Automation Technology Branch facilities and research is presented. Manipulator research includes dual-arm coordination studies, space manipulator dynamics, end-effector controller development, automatic space structure assembly, and the development of a dual-arm master-slave telerobotic manipulator system. Sensor research includes gravity-compensated force control, real-time monovision techniques, and laser ranging. Artificial intelligence techniques are being explored for supervisory task control, collision avoidance, and connectionist system architectures. A high-fidelity dynamic simulation of robotic systems, ROBSIM, is being supported and extended. Cooperative efforts with Oak Ridge National Laboratory have verified the ability of teleoperators to perform complex structural assembly tasks, and have resulted in the definition of a new dual-arm master-slave telerobotic manipulator. A bibliography of research results and a list of technical contacts are included

新規高感度定量法を用いた小胞体ストレス応答性因子 mesencephalic astrocyte-derived neurotrophic factor とその paralog の性状解析

岐阜大学(Gifu University)博士(薬科学)博士論文 (Doctoral dissertation)doctoral thesi

Electrical and Optical properties of GaSe-SnO_2 Heterojunctions

Capacitance-voltage (C-V) characteristics, current-voltage (I-V) characteristics, electroluminescence (E.L) and photovoltaic effect (I_) of GaSe-SnO_2 heterojunction diodes are measured.SnO_2 layer is deposited on the c-plane of GaSe by spray method. C-V characteristics of these diodes reveal that the high resistivity layer whose width is about 2.6 μm is formed. The origin of the high resistivity layer is probably due to the diffusion of Sn into GaSe.Forward I-V characteristics show the current transport mechanism at low voltage is space-charge-limited-current. The trap density and the energy level of the trap from the valence band estimated by Lampert theory are about 5x10^ -- 1x10^ cm^ and 0.4 -- 0.6 eV, respectively. E.L spectra at 275 K show only one emission band due to free exciton recombination. On the otherhand, E.L spectra at 4.2 K show many fine structures, and they are assigned to be due to bound excitons and its L.O. phonon replicas.III-VI族半導体GaSeはマイカ状の層状構造をしている。そのため電気的，光学的特性の異方性，特にエネルギー面の2次元性に興味が持たれ，多くの研究者によって実験的，理論的に研究されて来た。しかし，この結晶のエネルギー面は，その結晶構造の特異性にもかかわらず，やはり3次元的であることが最近明らかにされた。一方，GaSeの低温における発光スペクトルは多数の微細構造を示すが，その発光機構については末だ議論が多い。我々は，GaSeの発光機構を調べるため，GaSe-SnO_2ヘテロ接合を用いている。GaSeの禁制帯幅は，室温で2.03eVであるが，沃素を用いて閉管法で成長させた場合以外は常にp型である。又，SnO_2は3.5eVと広い禁制帯幅を持つn型半導体である。従って，GaSe-SnO_2接合をつくれば，p-nヘテロ接合となり，SnO_2からGaSeへの電子の注入が容易になると考えられる。SnO_2はスプレー法でGaSeのc面上に形成される。GaSeの裏面はInでオーミック電極をつける。この構造は，GaSeの同じc面上に2つの電極をつける従来の構造と比較して，かなり低電圧で発光するので，GaSeの発光機構を調べる上で有利である。本稿では，GaSe-SnO_2ヘテロ接合の作成と，その容量―電圧(C-V)特性，電流―電圧(I-V)特性，電界発光，光起電力効果の実験結果を報告し，電流輸送機構，発光機構について考察する

Scene illuminate effect on additive two-color reproduction

Whether a two-color reproduction from a red and green additive process is acceptable depends largely on the nature of the scene and its illumination. Additive two-color reproduction will be explored here using different scene illuminants and also different red and green filter combinations. Two projection methods are also examined. Red and green separations were produced by photographing a scene through 16 different filter combinations and 3 different scene illuminants. These separations are then projected two different ways. One method involves projecting the separations back through the red and green filters they were taken through, while in the other method the green filter is removed and replaced with a neutral density. It was found that tungsten was the best scene illuminant at reproducing the original scene colors, and that the best overall filter combination at doing this was a 26,61 Wratten filter combination

Spatiotemporal Expression of MANF in the Developing Rat Brain

Mesencephalic astrocyte-derived neurotrophic factor (MANF) is an evolutionarily conserved neurotrophic factor which exhibited neuroprotective properties. Recent studies suggested that MANF may play a role in the neural development of Drosophila and zebra fishes. In this study, we investigated the spatiotemporal expression of MANF in the brain of postnatal and adult rats. MANF expression appeared wide spread and mainly localized in neurons. In the cerebral cortex, neurons in layer IV and VI displayed particularly strong MANF immunoreactivity. In the hippocampus, intensive MANF expression was observed throughout the subfields of Cornu Amonis (CA1, CA2, and CA3) and the granular layer of the dentate gyrus (DG). In the substantia nigra, high MANF expression was shown in the substantia nigra pars compacta (SNpc). In the thalamus, the anterodorsal thalamic nucleus (ADTN) exhibited the highest MANF immunoreactivity. In the hypothalamus, intensive MANF immunoreactivity was shown in the supraoptic nucleus (SON) and tuberomammillary nucleus (TMN). In the cerebellum, MANF was localized in the external germinal layer (EGL), Purkinje cell layer (PCL), internal granule layer (IGL) and the deep cerebellar nuclei (DCN). We examined the developmental expression of MANF on postnatal day (PD) 3, 5, 7, 9, 15, 21, 30 and adulthood. In general, the levels of MANF were high in the early PDs (PD3 and PD5), and declined gradually as the brain matured; MANF expression in the adult brain was the lowest among all time points examined. However, in some structures, such as PCL, IGL, SON, TMN and locus coeruleus (LC), high expression of MANF sustained throughout the postnatal period and persisted into adulthood. Our results indicated that MANF was developmentally regulated and may play a role in the maturation of the central nervous system (CNS)

Hydrological Modeling of the Peruvian-Ecuadorian Amazon Basin Using GPM-IMERG Satellite-Based Precipitation Dataset

In the last two decades, rainfall estimates provided by the Tropical Rainfall Measurement Mission (TRMM) have proven applicable in hydrological studies. The Global Precipitation Measurement (GPM) mission, which provides the new generation of rainfall estimates, is now considered a global successor to TRMM. The usefulness of GPM data in hydrological applications, however, has not yet been evaluated over the Andean and Amazonian regions. This study uses GPM data provided by the Integrated Multi-satellite Retrievals (IMERG) (productfinal run) as input to a distributed hydrological model for the Amazon Basin of Peru and Ecuador for a 16-month period (from March 2014 to June 2015) when all datasets are available. TRMM products (TMPA V7, TMPA RT datasets) and a gridded precipitation dataset processed from observed rainfall are used for comparison. The results indicate that precipitation data derived from GPM-IMERG correspond more closely to TMPA V7 than TMPA RT datasets, but both GPM-IMERG and TMPA V7 precipitation data tend to overestimate, compared to observed rainfall (by 11.1 and 15.7 , respectively). In general, GPM-IMERG, TMPA V7 and TMPA RT correlate with observed rainfall, with a similar number of rain events correctly detected (20). Statistical analysis of modeled streamflows indicates that GPM-IMERG is as useful as TMPA V7 or TMPA RT datasets in southern regions (Ucayali basin). GPM-IMERG, TMPA V7 and TMPA RT do not properly simulate streamflows in northern regions (Maran and Napo basins), probably because of the lack of adequate rainfall estimates in northern Peru and the Ecuadorian Amazon

Modelling the evolutionarily conserved MANF/CDNF protein family in Drosophila melanogaster

Mesencephalic astrocyte-derived neurotrophic factor (MANF) and cerebral dopamine neurotrophic factor (CDNF) proteins form a family of neurotrophic factors. Neurotrophic factors have been intensively studied as a putative therapeutic approach to treat neuronal injuries and neurodegenerative diseases. Mammalian MANF and CDNF have been shown to have protective and restorative effects on the nigrostriatal dopaminergic system. In addition, several studies have reported a role for MANF in the endoplasmic reticulum (ER) stress response. A recently established MANF knockout mouse model revealed that MANF functions in the pancreatic insulin-producing beta cells and might be involved in the pathogenesis of diabetes mellitus. Beyond their neurotrophic properties, MANF and CDNF appear to play a more general role in the maintenance of cellular homeostasis. In this study, Drosophila melanogaster was used as a model organism to explore the function and interaction of the MANF/CDNF protein family in vivo. The sole member of the MANF/CDNF family in Drosophila, DmManf, was discovered to be crucial for fly development. The human orthologues, HsMANF and HsCDNF, were found to be able to substitute the endogenous DmManf. Likewise, DmManf had the cytoprotective properties of mammalian MANF in cultured murine neurons. These results support that the findings from the Drosophila model can be adapted for research in mammalian systems. MANF/CDNF proteins consist of amino (N) - and carboxy (C) -terminal domains. In this work, several functional features identified in mammalian MANF structure were explored in the Drosophila model. Separate N- or C-terminal domain constructs, even when co-expressed together, failed to complement for the loss of endogenous DmManf. The ER retention of DmManf, mediated by the C-terminal signal sequence, and the positive charge of the N-terminal surface amino acid residues were found to be important for appropriate DmManf function. Furthermore, entering the secretory pathway via ER was essential for the stability of DmManf protein. A CXXC motif characteristic for oxidoreductases is located in the C-terminal domain of MANF. In this study, effects of a point mutation (C129S) in CXXC motif of DmManf were analysed in vivo. Intact CXXC motif was discovered to be vital for DmManf function. Furthermore, the expression of DmManf-C129S in wild type background was harmful for fly viability suggesting that this specific mutation represents either a dominant negative or a gain-of-function allele of DmManf. Utilising the unique potential of Drosophila model for in vivo screening, interactions of DmManf were studied in this work. Consistent with a previous in vitro study, a genetic interaction was found between DmManf and the fly homologue of the major ER chaperone GRP78. Moreover, DmManf interacted with other genes that encode components of ER function and the unfolded protein response. Finally, novel interactions with DmManf and genes involved in the ubiquinone synthesis pathway and mitochondria were discovered. Taken together, this study demonstrates the functional conservation of mammalian and fly proteins and provides meaningful information on structural and functional features of the MANF/CDNF protein family in vivo. The genetic interaction studies confirmed and expanded the previous knowledge on the ER-associated functions of MANF. Furthermore, novel interactions with mitochondria-related genes and DmManf were discovered.Hermoston rappeutumissairaudet ovat yleistyneet ihmisten eliniän kohotessa. Tämänhetkisten hoitokeinojen avulla pystytään ainoastaan lievittämään hermoston rappeutumisesta aiheutuvia oireita eikä hermosolujen tuhoutumista voida pysäyttää. Yhtenä mahdollisena hoitomuotona tutkitaan erityisiä hermosolujen kasvua ja erilaistumista tukevia proteiineja, hermokasvutekijöitä. Hermokasvutekijöillä on tärkeä rooli yksilönkehityksen aikana hermoston muodostuessa, mutta niillä on merkittävä hermoston toimintaa ylläpitäviä tehtäviä myös aikuisiässä. MANF (mesencephalic astrocyte derived neurotrophic factor) ja CDNF (cerebral dopamine neurotrophic factor) muodostavat viimeisimpänä löydetyn hermokasvutekijäperheen. Nisäkkäiden MANF- ja CDNF-proteiinien on todettu edistävän dopamiinivälitteisten hermosolujen toimintaa ja jopa uudistavan niitä rotan Parkinsonin tautimallissa. Parkinsonin taudin oireet aiheutuvat dopamiinivälitteisten hermosolujen tuhoutumisesta ja näin ollen MANF- ja CDNF-proteiineja tutkitaan mahdollisena uutena hoitokeinona tähän sairauteen. MANF-proteiinilla tehdyt tutkimukset ovat osoittaneet, että MANF:lla on myös yleisempi rooli solujen tasapainon ylläpitämisessä, erityisesti solulimakalvoston stressitilassa. Hiirikokeissa on todettu MANF:n olevan korvaamattoman tärkeä insuliinia tuottavien haimasolujen toiminnalle ja MANF-poistogeenisille hiirille kehittyykin diabetes. Täten MANF- ja CDNF-proteiineja voidaan mahdollisesti hyödyntää myös diabeteksen hoidossa. Ennen kuin näitä proteiineja voidaan turvallisesti käyttää sairauksien hoidossa, on niiden toiminta tunnettava tarkasti mahdollisten haittavaikutusten tunnistamiseksi ja välttämiseksi. MANF/CDNF-perhe on ainoa hermokasvutekijäperhe, jolle löytyy vastine myös selkärangattomista eliöistä. Tässä työssä kartoitettiin MANF/CDNF-proteiiniperheen toimintaa elävässä mallieliössä, banaanikärpäsessä (Drosophila melanogaster), käyttäen sekä poistogeenisiä että siirtogeenisiä menetelmiä. Työn alkuvaiheissa kuvattiin banaanikärpäsen DmManf-proteiini ja sen ilmentyminen kärpäsen eri kehitysvaiheissa ja kudoksissa. DmManf-proteiinin puutos johti kärpästen kuolemaan varhaisessa kehitysvaiheessa ja sen huomattiin vähentävän dopamiinivälitteisten hermosolujen haarakkeiden määrää. Ihmisen MANF ja CDNF pystyivät korvaamaan kärpäsen DmManf:n puutoksen. Vastaavasti DmManf suojasi nisäkkäiden MANF:n tavoin hiiren viljeltyjä hermosoluja kemikaalein aiheutetulta solukuolemalta. Nämä tulokset osoittivat, että MANF/CDNF-proteiiniperheen toiminta on kehittynyt jo varhaisessa evoluution vaiheessa ja säilynyt samankaltaisena vuosimiljoonien ajan. Banaanikärpäsmallin monipuolisia siirtogeenimenetelmiä sekä kantakirjastoja hyödyntämällä tutkittiin DmManf-proteiinin rakenteellisia ominaisuuksia sekä DmManf-geenin vuorovaikutusta muiden geenien kanssa. DmManf-proteiinin kaksiosaisen rakenteen molempien palasten sekä pintavarauksen todettiin olevan oleellisia sen toiminnallisuuden kannalta. DmManf:n oikeanlaiselle toiminnalle oli myös tärkeää, että se ohjataan solulimakalvostolle. Lisäksi aiempien nisäkkäiden MANF-proteiinilla saatujen tutkimustulosten mukaisesti DmManf-geenin huomattiin vuorovaikuttavan GRP78-kaitsijaproteiinia tuottavan geenin kanssa. Työn viimeisessä osassa tunnistettiin DmManf:n kanssa vuorovaikuttavia geenejä, joista merkittävimpänä ryhmänä esiintyi mitokondrioiden toimintaan liittyvät geenit. Työ on kokonaisuudessaan tärkeä osa MANF/CDNF-proteiiniperheen perustutkimusta ja saadut löydökset selventävät näiden proteiinien toimintaa sekä mahdollistavat tarkempia jatkotutkimuksia lääkinnällistä käyttöä silmälläpitäen