DOCSLIB.ORG

Melanosome Morphologies in Murine Models of Hermansky–Pudlak

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Melanosome Morphologies in Murine Models of Hermansky–Pudlak PRIORITY PUBLICATION Melanosome Morphologies in Murine Models of Hermansky^Pudlak Syndrome Re£ect Blocks in Organelle Development Thuyen Nguyen, Edward K. Novak,w Maryam Kermani, Joachim Fluhr, Luanne L. Peters,n Richard T. Swank,w and Maria L. Wei Department of Dermatology,Veterans A¡airs Medical Center, University of California, San Francisco, CA, U.S.A.; nJackson Laboratory, Bar Harbor, ME and wDepartment of Molecular and Cellular Biology, Roswell Park Cancer Center, Bu¡alo, NY, U.S.A. Hermansky^Pudlak syndrome is an autosomal recessive fer. The Hermansky^Pudlak syndrome strains are disease characterized by pigment dilution and pro- classi¢ed into morphologic groups characterized by longed bleeding time. At least 15 mutant mouse strains the step at which melanosome biogenesis or transfer to have been classi¢ed as models of Hermansky^Pudlak keratinocytes is inhibited, with the cappuccino strain syndrome. Some of the genes are implicated in intracel- observed to be blocked at the earliest step and gunmetal lular vesicle tra⁄cking: budding, targeting, and secre- blocked at the latest step. We show that all Hermansky^ tion. Many of the Hermansky^Pudlak syndrome genes Pudlak syndrome mutant strains except gunmetal have remain uncharacterized and their functions are un- an increase in unpigmented or hypopigmented imma- known. Clues to the functions of these genes can be ture melanosomal forms, leading to the hypopigmen- found by analyzing the physiologic and cellular pheno- ted coat colors seen in these strains. In contrast, the types. Here we have examined the morphology of the hypopigmentation seen in the gunmetal strain is due melanosomes in the skin of 10 of the mutant mouse to the retention of melanosomes in melanocytes, and Hermansky^Pudlak syndrome strains by transmission ine⁄cient transfer into keratinocytes. Key words: mela- electron microscopy.We demonstrate that the morphol- nocytes/organelle biogenesis/pigmentation J Invest Dermatol ogies re£ect inhibition of organelle maturation or trans- 119: 1156 ^1164,2002 he Hermansky^Pudlak syndrome (HPS) is observed subunits of the adaptor complex AP3 (Kantheti et al,1998; in humans and mice and is caused by mutations Dell’Angelica et al, 1999; Feng et al, 1999), which binds to endo- in genes that regulate the biogenesis of lysosomes somes and the trans-Golginetwork. A neuronal-speci¢c isoform and lysosome-related organelles such as melanosomes of AP3 is required for synaptic vesicle formation from endosomes and platelet-dense granules (Dell’Angelica et al,2000), (Faundez et al,1998).Pldn (palladin) protein binds syntaxin 13 Twhich suggests that these seemingly disparate organelles share a (Huang et al, 1999), a member of the family of soluble N-ethylma- common pathway of biogenesis or maturation. The resulting de- leimide-sensitive factor attachment protein receptors (SNARES), fects in these organelles cause prolonged bleeding and oculocuta- which mediate fusion of intracellular membranes. Rabgtta encodes neous albinism in a¡ected individuals (Huizing et al,2000).HPS a subunit of a Rab geranylgeranyl transferase (Detter et al,2000), exhibits genetic heterogeneity: in humans, four genes have been and is required for e⁄cient polarization of cytolytic T cell gran- cloned, and each when defective in isolation gives rise to a clinical ules (which are also specialized lysosome-like organelles) to the syndrome classi¢ed as HPS. In mice, at least 15 HPS genes have immunologic synapse (Stinchcombe et al,2001).Rab27a encodes been noted (Swank et al,1998). a member of the family of GTPases that controls intracellular Whereas the functions of many of the HPS gene products re- vesicular transport (Novick and Zerial, 1997; Chavrier and Goud, main unknown, it is likely that most of them will function in 1999) and is required for melanosome transport from the cell vesicle/organelle biogenesis and membrane tra⁄cking along the body to the dendritic processes (Wilson et al, 2000; Bahadoran pathway of melanosome formation. The functions of ¢ve genes et al, 2001; Hume et al, 2001). In mice with defective HPS1, (Ap3b1, Ap3d, Pldn, Rabgtta, Rab27a) have been implicated in ve- HPS3, HPS4, Pldn,ormu genes, melanosomes in eye tissue have sicle/organelle formation and tra⁄cking. Ap3b1 and Ap3d encode been observed to be abnormal (Ito et al, 1982; Gardner et al,1997; Suzuki et al, 2001, 2002; Zhang et al, 2002a). Eukaryotic cells are distinguished by the presence of mem- Manuscript received August 18, 2002; revised September 6, 2002; ac- brane-bound organelles. The study of organelle biogenesis has cepted for publication September 12, 2002 been greatly advanced by the study of organisms such as Sacchar- Reprint requests to: Dr Maria L.Wei, Dermatology Service (190), Veter- ans A¡airs Medical Center, 4150 Clement Street, San Francisco, CA 94121, omyces cerevisiae and Drosophila melanogaster that exhibit mutations U.S.A. Email: [email protected] in genes that regulate organelle development (Dell’Angelica et al, Abbreviations: HPS, Hermansky^Pudlak syndrome; TEM, transmission 2000). Morphologic studies of yeast cells bearing mutations in electron microscopy; Tyrp1/TRP-1, tyrosinase-related protein 1; TRP-2, genes that govern the biogenesis of the yeast vacuole (an orga- tyrosinase-related protein 2. nelle analogous to the mammalian lysosome) have helped to 0022-202X/02/$15.00 Copyright r 2002 by The Society for Investigative Dermatology, Inc. 1156 VOL. 119, NO. 5 NOVEMBER 2002 MELANOSOMES IN HERMANSKY^PUDLAK SYNDROME 1157 identify and classify more than 40 genes that were involved (Novak et al, 1980, 1984; Gibb et al, 1981; Zhang et al, 2002b).The cappuccino (Banta et al, 1988; Raymond et al, 1992;Wada et al, 1992). Although strain arose on the C3H/HeJ background (Gwynn et al, 2000). The cocoa the human HPS1, HPS3,andHPS4 genes have no sequence mouse arose on the C57BL/10J background (Novak et al, 1988). The three homologs in yeast, the HPS2/Ap3b1 gene has a yeast homolog control strains had very similar melanosome morphology (data not shown), so C57BL/6 was used as a representative control. The HPS mice and some of the as yet uncharacterized mouse genes may have examined and the corresponding human diseases and genes are listed in yeast homologs as well. HPS mutations result in mammalian dis- Tabl e I . All animal procedures were approved by Institutional Animal eases due to defects in organelle biogenesis. We have undertaken Care Use Committee. the current morphologic study in order to characterize and classi- fy HPS mouse strains as a step towards understanding HPS gene Measurement of coat color The dorsal back coat pigmentation (P) was product function. measured using a Mexameter MX 18 (Courage & Khazaka, Cologne, The melanosome is an organelle uniquely suited for morpho- Germany), which gives a calculated value of melanin content based on logic examination in tissue samples by transmission electron mi- the absorption of light emitted at 568 nm and 880 nm. Background was subtracted by subtracting Ptyr, the coat color measurement for the white croscopy (TEM) due to its electron-dense pigmentation. Other albino mouse C57BL/6J-Ty r c^2J (TyrG291 T), which is lacking tyrosinase organelles consistently a¡ected in HPS are not readily amenable activity and has no pigment production. A comparison with the coat to examination by TEM. Platelet-dense granules in HPS lose pigment of the control strain (Pcon) was done: their electron density (Witkop et al, 1987) and lysosomes are only easily identi¢ed after internalization of electron-dense material or Percent pigment ¼ðP À PtyrÞ=ðPcon À PtyrÞ100 by antibody labeling, and thus are more di⁄cult to identify in tissue samples. Electron microscopy Tissue was ¢xed in modi¢ed Karnovsky’s ¢xative The melanosome has four distinct stages of maturation (Nordlund (2% paraformaldehyde/2% glutaraldehyde/0.1 M cacodylate bu¡er, pH 7.3/ et al, 1998), stages I^IV.Type I melanosomes have intralumenal vesi- 0.06% CaCl2) for 24 h, washed twice in 0.1 M cacodylate bu¡er, post¢xed cles (Raposo et al, 2001) and resemble multivesicular bodies, struc- in reduced osmium (1.5% potassium ferrocyanide/2% osmium tetroxide) tures found along the endosomal pathway. Type II is characterized for 2 h, rinsed in H2O, dehydrated with increasing ethanol concentrations, by an elongated, elliptical shape, with intralumenal ¢ne ¢brils giv- and embedded in Epon resin. Grids were stained in 10% uranyl acetate/ ing a striated appearance; type III exhibits pigment deposition along 50% methanol. Cross-sections of melanosomes were measured at  50,000 magni¢cation for major axis and minor axis, in order to assess the ¢brils, and type IV has dense pigmentation ¢lling the organelle relative size and shape. Images of a calibration grid were taken to control and obscuring the ¢brillar structure (Seiji et al, 1963). In normal mel- for any variability in magni¢cation. Melanocytes were identi¢ed by their anosome development, the type I melanosome is distinguished by organization in hair follicles, as well as their lack of keratin ¢laments and its accumulation of the Pmel17/gp100 protein (silver locus product) desmosomes (both are present in keratinocytes). Each melanosome was (Kushimoto et al,2001;Raposoet al, 2001; Raposo and Marks, measured and classi¢ed by morphology, i.e.: (i) multiple intralumenal 2002), which is cleaved and localized to the intralumenal vesicles vesicles present, no pigment or striations seen; (ii) striated matrix present, (Berson et al, 2001). Two enzymes involved in pigment synthesis, round cross-section; (iii) striated matrix, elliptical cross-section; (iv) fully Tyrp1/ tyrosinase-related protein (TRP)-1 and dopachrome tauto- pigmented, round cross-section; and (v) fully pigmented, elliptical cross- merase/TRP-2, may also be in the type I melanosome, but appear section. Noted also were any melanosomal features unique to any particular strain. For each strain at least 100 melanosomes were examined to be inactivated in this compartment by proteolytic cleavage (Kush- in each of at least two di¡erent animals.
Load more

Recommended publications
  • Heřmanský–Pudlák Syndrome: a Case Report
    Int J Case Rep Images 2018;9:100886Z01SB2018. Basu et al. 1 www.ijcasereportsandimages.com CASE REPORT PEER REVIEWED | OPEN ACCESS Heřmanský–Pudlák syndrome: A case report Sonali Someek Basu, Pranav Kumar ABSTRACT care, rehabilitation and multidisciplinary team approach. Heřmanský–Pudlák syndrome (HPS) is hereditary multi system, a rare autosomal Keywords: Gene and data base, Heřmanský– recessive disorder with heterogeneous Pudlák syndrome, Multiple skin cancer, Oculocu- locus characterized by tyrosine positive taneous albinism, Pulmonary fibrosis oculocutaneous albinism, congenital nystagmus, platelet storage pool deficiency - How to cite this article hemorrhagic bleeding tendency and systemic manifestations due to lysosomal accumulation Basu SS, Kumar P. Heřmanský–Pudlák of ceroid lipofuscin in different organs includes syndrome: A case report. Int J Case Rep Images pulmonary fibrosis and granulomatous 2018;9:100886Z01SB2018. enteropathy colitis and renal failure. Depending upon the organ system symptoms and signs, the presentation of HPS can be varied and diagnosis Article ID: 100886Z01SB2018 can be challenging. The most frequently used criteria for diagnosis of HPS as per National Organization of Rare Disease (NORD) revised ********* 2015 are albinism and prolong bleeding. doi: 10.5348/100886Z01SB2018CR Symptoms due to ceroid lipofuscin deposition in lungs, colon, heart and kidneys can occur. Differentiating this condition from other diseases that cause oculocutaneous albinism INTRODUCTION such as ocular albinism, Griscelli syndrome and Chédiak–Higashi syndrome by presence of Heřmanský–Pudlák syndrome (HPS), a rare neurological symptoms and frequent bacterial hereditary disease caused by autosomal recessive infections due to immunodeficiency and analysis inheritance, was first described by Hermansky and of peripheral blood smear with prolong bleeding Pudlakin 1959 [1].
    [Show full text]
  • A Gene for Freckles Maps to Chromosome 4Q32–Q34
    A Gene for Freckles Maps to Chromosome 4q32–q34 Xue-Jun Zhang,Ãz1 Ping-Ping He,Ãwz Yan-Hua Liang,Ãwz Sen Yang,Ãz Wen-Tao Yuan,w Shi-Jie Xu,w and Wei Huangw1 ÃInstitute of Dermatology & Department of Dermatology at No.1 Hospital, Anhui Medical University, Hefei, China, wChinese National Human Genome Center at Shanghai, Shanghai, China, zKey Laboratory of Genome Research at Anhui, Hefei, China Freckles are numerous pigmented macules on the face commonly occurring in the Caucasian and Chinese population. As freckling is considered as an independent trait, no gene or locus for it has been identified to date. Here we performed genome-wide scan for linkage analysis in a multigeneration Chinese family with freckles. A maximum LOD score of 4.26 at a recombination fraction of 0 has been obtained with marker D4S1566. Haplotype analysis localized the freckles locus to a 16 Mbp region flanked by D4S2952 and D4S1607. We have thus mapped the gene for freckles to chromosome 4q32–q34. This will aid future identification of the responsible gene, which will be very useful for the understanding of the molecular mechanism of freckles. Key words: freckles/gene mapping/genome-wide scan/linkage analysis. J Invest Dermatol 122:286 –290, 2004 Freckles, or ephelides are numerous pigmented spots of the tion study with the melanocorin-1-receptor (MC1R) gene, skin commonly occurring in the Caucasian population freckles, and solar lentigines, and found that the MC1R (Bastiaens et al, 1999). They are mainly confined to the gene variants could play an important part in the develop- face, even arms and back.
    [Show full text]
  • Supplementary Table 1 Final.Xlsx
    Platelet Biology & Its Disorders SUPPLEMENTARY APPENDIX Whole exome sequencing identifies genetic variants in inherited thrombocytopenia with secondary qualitative function defects Ben Johnson, 1 Gillian C. Lowe, 1 Jane Futterer, 1 Marie Lordkipanidzé, 1 David MacDonald, 1 Michael A. Simpson, 2 Isabel Sanchez-Guiú, 3 Sian Drake, 1 Danai Bem, 1 Vincenzo Leo, 4 Sarah J. Fletcher, 1 Ban Dawood, 1 José Rivera, 3 David Allsup, 5 Tina Biss, 6 Paula HB Bolton-Maggs, 7 Peter Collins, 8 Nicola Curry, 9 Charlotte Grimley, 10 Beki James, 11 Mike Makris, 4 Jayashree Motwani, 12 Sue Pavord, 13 Katherine Talks, 6 Jecko Thachil, 7 Jonathan Wilde, 14 Mike Williams, 12 Paul Harrison, 15 Paul Gis - sen, 16 Stuart Mundell, 17 Andrew Mumford, 18 Martina E. Daly, 4 Steve P. Watson, 1 and Neil V. Morgan 1 on behalf of the UK GAPP Study Group 1Institute for Cardiovascular Sciences, College of Medical and Dental Sciences, University of Birmingham, UK; 2Division of Genetics and Molecular Medicine, King's College, London, UK; 3Centro Regional de Hemodonación, Universidad de Murcia, IMIB-Arrixaca, Mur - cia, Spain; 4Department of Infection, Immunity and Cardiovascular Disease, University of Sheffield Medical School, University of Sheffield, UK; 5Hull Haemophilia Treatment Centre, Hull and East Yorkshire Hospitals NHS Trust, Castle Hill Hospital, Hull, UK; 6Depart - ment of Haematology, Royal Victoria Infirmary, Newcastle Upon Tyne, UK; 7Department of Haematology, Manchester Royal Infirmary, Manchester, UK; 8Arthur Bloom Haemophilia Centre, School of Medicine, Cardiff
    [Show full text]
  • Development of the Swimbladder Surfactant System and Biogenesis of Lysosome-Related Organelles Is Regulated by BLOS1 in Zebrafish
    | INVESTIGATION Development of the Swimbladder Surfactant System and Biogenesis of Lysosome-Related Organelles Is Regulated by BLOS1 in Zebrafish Tianbing Chen,*,† Guili Song,* Huihui Yang,*,† Lin Mao,*,† Zongbin Cui,*,1 and Kaiyao Huang*,1 *Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China and †University of Chinese Academy of Sciences, Beijing 100049, China ABSTRACT Hermansky-Pudlak syndrome (HPS) is a human autosomal recessive disorder that is characterized by oculocutaneous albinism and a deficiency of the platelet storage pool resulting from defective biogenesis of lysosome-related organelles (LROs). To date, 10 HPS genes have been identified, three of which belong to the octamer complex BLOC-1 (biogenesis of lysosome-related organelles complex 1). One subunit of the BLOC-1 complex, BLOS1, also participates in the BLOC-1-related complex (BORC). Due to lethality at the early embryo stage in BLOS1 knockout mice, the function of BLOS1 in the above two complexes and whether it has a novel function are unclear. Here, we generated three zebrafish mutant lines with a BLOC-1 deficiency, in which melanin and silver pigment formation was attenuated as a result of mutation of bloc1s1, bloc1s2, and dtnbp1a, suggesting that they function in the same complex. In addition, mutations of bloc1s1 and bloc1s2 caused an accumulation of clusters of lysosomal vesicles at the posterior part of the tectum, representing a BORC-specific function in zebrafish. Moreover, bloc1s1 is highly expressed in the swimbladder during postembryonic stages and is required for positively regulating the expression of the genes, which is known to govern surfactant production and lung development in mammals.
    [Show full text]
  • MUTED Antibody - C-Terminal Region (ARP63301 P050) Data Sheet
    MUTED antibody - C-terminal region (ARP63301_P050) Data Sheet Product Number ARP63301_P050 Product Name MUTED antibody - C-terminal region (ARP63301_P050) Size 50ug Gene Symbol BLOC1S5 Alias Symbols DKFZp686E2287; MU; MUTED Nucleotide Accession# NM_201280 Protein Size (# AA) 187 amino acids Molecular Weight 21kDa Product Format Lyophilized powder NCBI Gene Id 63915 Host Rabbit Clonality Polyclonal Official Gene Full Name Muted homolog (mouse) Gene Family BLOC1S This is a rabbit polyclonal antibody against MUTED. It was validated on Western Blot by Aviva Systems Biology. At Aviva Systems Biology we manufacture rabbit polyclonal antibodies on a large scale (200-1000 Description products/month) of high throughput manner. Our antibodies are peptide based and protein family oriented. We usually provide antibodies covering each member of a whole protein family of your interest. We also use our best efforts to provide you antibodies recognize various epitopes of a target protein. For availability of antibody needed for your experiment, please inquire (). Partner Proteins BLOC1S2,DTNBP1,BLOC1S1,BLOC1S2,CNO,DTNBP1,SNAPIN,BLOC1S2,BLOC1S3,DTNBP1,PLDN,SQS TM1,YOD1 This gene encodes a component of BLOC-1 (biogenesis of lysosome-related organelles complex 1). Components of this complex are involved in the biogenesis of organelles such as melanosomes and platelet- Description of Target dense granules. A mouse model for Hermansky-Pudlak Syndrome is mutated in the murine version of this gene. Alternative splicing results in multiple transcript variants. Read-through transcription exists between this gene and the upstream EEF1E1 (eukaryotic translation elongation factor 1 epsilon 1) gene, as well as with the downstream TXNDC5 (thioredoxin domain containing 5) gene.
    [Show full text]
  • Product Information
    Product information PLDN, 1-172aa Human, His-tagged, Recombinant, E.coli Cat. No. IBATGP1472 Full name: pallidin NCBI Accession No.: NP_036520 Synonyms: HPS9, PA, PALLID Description: PLDN (Pallidin) may play a role in intracellular vesicle trafficking. It interacts with Syntaxin 13 which mediates intracellular membrane fusion. Several alternatively spliced transcript variants of this gene have been described, but the full-length nature of some of these variants has not been determined. This protein involved in the development of lysosome-related organelles, such as melanosomes and platelet-dense granules. PLDN has been shown to interact with BLOC1S1, STX12, Dysbindin, CNO, BLOC1S2, MUTED and SNAPAP. Recombinant human PLDN protein, fused to His-tag at N-terminus, was expressed in E.coli and purified by using conventional chromatography techniques. Form: Liquid. In 20mM Tris-HCl buffer (pH 8.0) containing 2mM DTT, 10% glycerol, 100mM NaCl Molecular Weight: 21.9kDa (192aa), confirmed by MALDI-TOF (Molecular weight on SDS-PAGE will appear higher) Purity: > 90% by SDS - PAGE Concentration: 1mg/ml (determined by Bradford assay) 15% SDS-PAGE (3ug) Sequences of amino acids: MGSSHHHHHH SSGLVPRGSH MSVPGPSSPD GALTRPPYCL EAGEPTPGLS DTSPDEGLIE DLTIEDKAVE QLAEGLLSHY LPDLQRSKQA LQELTQNQVV LLDTLEQEIS KFKECHSMLD INALFAEAKH YHAKLVNIRK EMLMLHEKTS KLKKRALKLQ QKRQKEELER EQQREKEFER EKQLTARPAK RM General references: Huang L, et al. (1999). Nat Genet 23 (3): 329–32. Moriyama K., et al. (2002) Traffic 3:666-677 Storage: Can be stored at +4°C short term (1-2 weeks). For long term storage, aliquot and store at -20°C or -70°C. Avoid repeated freezing and thawing cycles. For research use only. This product is not intended or approved for human, diagnostics or veterinary use.
    [Show full text]
  • E-Mutpath: Computational Modelling Reveals the Functional Landscape of Genetic Mutations Rewiring Interactome Networks
    bioRxiv preprint doi: https://doi.org/10.1101/2020.08.22.262386; this version posted August 24, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. e-MutPath: Computational modelling reveals the functional landscape of genetic mutations rewiring interactome networks Yongsheng Li1, Daniel J. McGrail1, Brandon Burgman2,3, S. Stephen Yi2,3,4,5 and Nidhi Sahni1,6,7,8,* 1Department oF Systems Biology, The University oF Texas MD Anderson Cancer Center, Houston, TX 77030, USA 2Department oF Oncology, Livestrong Cancer Institutes, Dell Medical School, The University oF Texas at Austin, Austin, TX 78712, USA 3Institute For Cellular and Molecular Biology (ICMB), The University oF Texas at Austin, Austin, TX 78712, USA 4Institute For Computational Engineering and Sciences (ICES), The University oF Texas at Austin, Austin, TX 78712, USA 5Department oF Biomedical Engineering, Cockrell School of Engineering, The University oF Texas at Austin, Austin, TX 78712, USA 6Department oF Epigenetics and Molecular Carcinogenesis, The University oF Texas MD Anderson Science Park, Smithville, TX 78957, USA 7Department oF BioinFormatics and Computational Biology, The University oF Texas MD Anderson Cancer Center, Houston, TX 77030, USA 8Program in Quantitative and Computational Biosciences (QCB), Baylor College oF Medicine, Houston, TX 77030, USA *To whom correspondence should be addressed. Nidhi Sahni. Tel: +1 512 2379506; Email: [email protected] 1 bioRxiv preprint doi: https://doi.org/10.1101/2020.08.22.262386; this version posted August 24, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder.
    [Show full text]
  • Gene Expression in the Mouse Eye: an Online Resource for Genetics Using 103 Strains of Mice
    Molecular Vision 2009; 15:1730-1763 <http://www.molvis.org/molvis/v15/a185> © 2009 Molecular Vision Received 3 September 2008 | Accepted 25 August 2009 | Published 31 August 2009 Gene expression in the mouse eye: an online resource for genetics using 103 strains of mice Eldon E. Geisert,1 Lu Lu,2 Natalie E. Freeman-Anderson,1 Justin P. Templeton,1 Mohamed Nassr,1 Xusheng Wang,2 Weikuan Gu,3 Yan Jiao,3 Robert W. Williams2 (First two authors contributed equally to this work) 1Department of Ophthalmology and Center for Vision Research, Memphis, TN; 2Department of Anatomy and Neurobiology and Center for Integrative and Translational Genomics, Memphis, TN; 3Department of Orthopedics, University of Tennessee Health Science Center, Memphis, TN Purpose: Individual differences in patterns of gene expression account for much of the diversity of ocular phenotypes and variation in disease risk. We examined the causes of expression differences, and in their linkage to sequence variants, functional differences, and ocular pathophysiology. Methods: mRNAs from young adult eyes were hybridized to oligomer microarrays (Affymetrix M430v2). Data were embedded in GeneNetwork with millions of single nucleotide polymorphisms, custom array annotation, and information on complementary cellular, functional, and behavioral traits. The data include male and female samples from 28 common strains, 68 BXD recombinant inbred lines, as well as several mutants and knockouts. Results: We provide a fully integrated resource to map, graph, analyze, and test causes and correlations of differences in gene expression in the eye. Covariance in mRNA expression can be used to infer gene function, extract signatures for different cells or tissues, to define molecular networks, and to map quantitative trait loci that produce expression differences.
    [Show full text]
  • BLOS2 Negatively Regulates Notch Signaling During Neural and Hematopoietic Stem And
    1 BLOS2 negatively regulates Notch signaling during neural and hematopoietic stem and 2 progenitor cell development 3 4 Wenwen Zhou 1, 6 #, Qiuping He 2, 6 #, Chunxia Zhang 2, 6, Xin He 1, Zongbin Cui 3, 5 Feng Liu 2, 6 *, and Wei Li 1, 4, 5 * 6 7 1 State Key Laboratory of Molecular Developmental Biology, Institute of Genetics and 8 Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China; 9 2 State Key Laboratory of Membrane Biology, Institute of Zoology, Chinese Academy of 10 Sciences, Beijing 100101, China 11 3 State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, 12 Chinese Academy of Sciences, Wuhan 430072, China 13 4 Center for Medical Genetics, Beijing Children’s Hospital, Capital Medical University; 14 Beijing Pediatric Research Institute; MOE Key Laboratory of Major Diseases in Children, 15 Beijing 100045, China; 16 5 Center of Alzheimer's Disease, Beijing Institute for Brain Disorders, Beijing 100053, China; 17 6 University of Chinese Academy of Sciences, Beijing 100039, China. 18 19 # Co-first authors. 20 * Correspondence should be addressed to: 21 Wei Li, Ph.D., Beijing Children’s Hospital, Capital Medical University, 56 Nan-Li-Shi Road, 1 22 Xicheng District, Beijing 100045, China. Tel: +86-10-5961-6628; Fax: +86-5971-8699; 23 e-mail: [email protected]; 24 Feng Liu, Ph.D., State Key Laboratory of Membrane Biology, Institute of Zoology, Chinese 25 Academy of Sciences, Beijing 100101, China. Tel.: +86 (10) 64807307; Fax: +86 (10) 26 64807313; Email: [email protected]. 27 28 Running Title: BLOS2 negatively regulates Notch signaling 29 30 Key Words: BLOS2; Notch; stem and progenitor cell development; neurogenesis; 31 hematopoiesis; endo-lysosomal trafficking 32 33 The authors declare that no competing interests exist.
    [Show full text]
  • Downregulation of Carnitine Acyl-Carnitine Translocase by Mirnas
    Page 1 of 288 Diabetes 1 Downregulation of Carnitine acyl-carnitine translocase by miRNAs 132 and 212 amplifies glucose-stimulated insulin secretion Mufaddal S. Soni1, Mary E. Rabaglia1, Sushant Bhatnagar1, Jin Shang2, Olga Ilkayeva3, Randall Mynatt4, Yun-Ping Zhou2, Eric E. Schadt6, Nancy A.Thornberry2, Deborah M. Muoio5, Mark P. Keller1 and Alan D. Attie1 From the 1Department of Biochemistry, University of Wisconsin, Madison, Wisconsin; 2Department of Metabolic Disorders-Diabetes, Merck Research Laboratories, Rahway, New Jersey; 3Sarah W. Stedman Nutrition and Metabolism Center, Duke Institute of Molecular Physiology, 5Departments of Medicine and Pharmacology and Cancer Biology, Durham, North Carolina. 4Pennington Biomedical Research Center, Louisiana State University system, Baton Rouge, Louisiana; 6Institute for Genomics and Multiscale Biology, Mount Sinai School of Medicine, New York, New York. Corresponding author Alan D. Attie, 543A Biochemistry Addition, 433 Babcock Drive, Department of Biochemistry, University of Wisconsin-Madison, Madison, Wisconsin, (608) 262-1372 (Ph), (608) 263-9608 (fax), [email protected]. Running Title: Fatty acyl-carnitines enhance insulin secretion Abstract word count: 163 Main text Word count: 3960 Number of tables: 0 Number of figures: 5 Diabetes Publish Ahead of Print, published online June 26, 2014 Diabetes Page 2 of 288 2 ABSTRACT We previously demonstrated that micro-RNAs 132 and 212 are differentially upregulated in response to obesity in two mouse strains that differ in their susceptibility to obesity-induced diabetes. Here we show the overexpression of micro-RNAs 132 and 212 enhances insulin secretion (IS) in response to glucose and other secretagogues including non-fuel stimuli. We determined that carnitine acyl-carnitine translocase (CACT, Slc25a20) is a direct target of these miRNAs.
    [Show full text]
  • Human BLOC1S2 / BLOS2 Protein (GST Tag)
    Human BLOC1S2 / BLOS2 Protein (GST Tag) Catalog Number: 13916-H09E General Information SDS-PAGE: Gene Name Synonym: BLOS2; CEAP; CEAP11 Protein Construction: A DNA sequence encoding the human BLOC1S2 (Q6QNY1-2) (Met1- Arg99) was fused with the GST tag at the N-terminus. Source: Human Expression Host: E. coli QC Testing Purity: > 74 % as determined by SDS-PAGE Endotoxin: Protein Description Please contact us for more information. BLOC1S2, also known as BLOS2, belongs to the BLOC1S2 family. It is a Stability: component of BLOC-1 complex. The BLOC-1 complex is composed of BLOC1S1, BLOC1S2, BLOC1S3, DTNBP1, MUTED, PLDN, Samples are stable for up to twelve months from date of receipt at -70 ℃ CNO/cappuccino and SNAPIN. The BLOC-1 complex is required for normal biogenesis of lysosome-related organelles, such as platelet dense Predicted N terminal: Met granules and melanosomes. BLOC1S2 interacts directly with BLOC1S1, Molecular Mass: BLOC1S3, MUTED, CNO/cappuccino and SNAPIN. It may play a role in cell proliferation. It also plays a role in intracellular vesicle trafficking. The recombinant human BLOC1S2 /GST chimera consists of 333 amino Functionally, BLOC1S2 gene has been proposed to participate in acids and has a predicted molecular mass of 38.7 kDa. It migrates as an processes (melanosome organization, microtubule nucleation, platelet approximately 39 KDa band in SDS-PAGE under reducing conditions. dense granule organization, positive regulation of cell proliferation, positive regulation of transcription, regulation of apoptosis, positive regulation of Formulation: transcription from RNA polymerase II promoter). Lyophilized from sterile PBS, pH 7.4 References Normally 5 % - 8 % trehalose, mannitol and 0.01% Tween80 are added as 1.Sowa ME, et al.
    [Show full text]
  • A BLOC-1 Mutation Screen Reveals That PLDN Is Mutated in Hermansky-Pudlak Syndrome Type 9
    CORE Metadata, citation and similar papers at core.ac.uk Provided by Elsevier - Publisher Connector ARTICLE A BLOC-1 Mutation Screen Reveals that PLDN Is Mutated in Hermansky-Pudlak Syndrome Type 9 Andrew R. Cullinane,1,* James A. Curry,1 Carmelo Carmona-Rivera,1 C. Gail Summers,2 Carla Ciccone,1 Nicholas D. Cardillo,1 Heidi Dorward,1 Richard A. Hess,1 James G. White,3 David Adams,1,4 Marjan Huizing,1 and William A. Gahl1,4 Hermansky-Pudlak Syndrome (HPS) is an autosomal-recessive condition characterized by oculocutaneous albinism and a bleeding diath- esis due to absent platelet delta granules. HPS is a genetically heterogeneous disorder of intracellular vesicle biogenesis. We first screened all our patients with HPS-like symptoms for mutations in the genes responsible for HPS-1 through HPS-6 and found no functional muta- tions in 38 individuals. We then examined all eight genes encoding the biogenesis of lysosome-related organelles complex-1, or BLOC-1, proteins in these individuals. This identified a homozygous nonsense mutation in PLDN in a boy with characteristic features of HPS. PLDN is mutated in the HPS mouse model pallid and encodes the protein pallidin, which interacts with the early endosomal t-SNARE syntaxin-13. We could not detect any full-length pallidin in our patient’s cells despite normal mRNA expression of the mutant transcript. We could detect an alternative transcript that would skip the exon that harbored the mutation, but we demonstrate that if this transcript is translated into protein, although it correctly localizes to early endosomes, it does not interact with syntaxin-13.
    [Show full text]