DOCSLIB.ORG

Loss of Hfe Leads to Progression of Tumor Phenotype in Primary Retinal Pigment Epithelial Cells

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Loss of Hfe Leads to Progression of Tumor Phenotype in Primary Retinal Pigment Epithelial Cells Retina Loss of Hfe Leads to Progression of Tumor Phenotype in Primary Retinal Pigment Epithelial Cells Jaya P. Gnana-Prakasam,1 Rajalakshmi Veeranan-Karmegam,1 Veena Coothankandaswamy,1 Sushma K. Reddy,1 Pamela M. Martin,1 Muthusamy Thangaraju,1 Sylvia B. Smith,2 and Vadivel Ganapathy1 PURPOSE. Hemochromatosis is a disorder of iron overload ereditary hemochromatosis is an autosomal recessive arising mostly from mutations in HFE. HFE is expressed in Hdisorder of iron overload.1–5 Excess iron is toxic because retinal pigment epithelium (RPE), and HfeÀ/À mice develop age- it can undergo Fenton reaction, catalyzing the conversion of related iron accumulation and retinal degeneration associated H2O2 to hydroxyl radical. Hydroxyl radicals in turn cause lipid with RPE hyperproliferation. Here, the mechanism underlying peroxidation, DNA strand breaks, and degradation of cellular the hyperproliferative phenotype in RPE was investigated. components resulting in tissue damage.6 Hemochromatosis METHODS. Cellular senescence was monitored by b-galactosi- patients have morbid iron accumulation in various organs, dase activity. Gene expression was monitored by real-time PCR. including the liver, pancreas, kidney, heart, and brain resulting Survivin was analyzed by Western blot and immunofluores- in diverse symptoms such as hepatocarcinoma/cirrhosis, cence. Migration and invasion were monitored using appro- diabetes, nephropathy, cardiomyopathy, and pituitary dysfunc- priate kits. Glucose transporters (GLUTs) were monitored by 3- tion.1–3 Most (>85%) cases of hemochromatosis are associated O-methyl-D-glucose uptake. Histone deacetylases (HDACs) with mutations in HFE [Histocompatability leukocyte antigen were studied by monitoring catalytic activity and acetylation class I-like protein involved in iron (FE) homeostasis].7 The status of histones H3/H4. clinical symptoms of the disease caused by mutations in HFE À/À begin to appear only at relatively older ages (>50 years). The RESULTS. Hfe RPE cells exhibited slower senescence rate and remaining approximately 15% of mutations occur in hepcidin, higher survivin expression than wild type cells. À/À cells Hfe hemojuvelin (HJV; also known as HFE2), ferroportin, and migrated faster and showed greater glucose uptake and transferrin receptor 2 (TfR2), all of which are also important increased expression of GLUTs. The expression of HDACs determinants of iron homeostasis.8 Mutations in HJV and and DNA methyltransferase (DNMTs) also was increased. hepcidin lead to iron overload at a much younger age, resulting Similarly, RPE cells from hemojuvelin (Hjv)-knockout mice, in juvenile hemochromatosis. HFE interacts with b -micro- another model of hemochromatosis, also had increased 2 globulin, and this interaction is obligatory for the presentation expression of GLUTs, HDACs, and DNMTs. The expression of of HFE to the cell surface. In the plasma membrane, HFE Slc5a8 was decreased in HfeÀ/À RPE cells, but treatment with a DNA methylation inhibitor restored the transporter expression, interacts with transferrin receptors, TfR1 and TfR2, and indicating involvement of DNA methylation in the silencing of inhibits cellular iron uptake. Irrespective of the gene that is Slc5a8 in HfeÀ/À cells. mutated in hemochromatosis, there is an increase in intestinal absorption of iron, leading to systemic iron overload. The CONCLUSIONS. RPE cells from iron-overloaded mice exhibit prevalence of hemochromatosis is quite high, with homozy- several features of tumor cells: decreased senescence, gosity for HFE mutations in the range of approximately 1 in enhanced migration, increased glucose uptake, and elevated 300.7 À/À levels of HDACs and DNMTs. These features are seen in Hfe In the retina, there are many iron-containing proteins that À/À RPE cells as well as in Hjv RPE cells, providing a molecular are involved in the phototransduction cascade9,10; hence, it is À/À À/À basis for the hyperproliferative phenotype of Hfe and Hjv essential to maintain iron homeostasis for effective functioning RPE cells. (Invest Ophthalmol Vis Sci. 2013;54:63–71) DOI: of the retina. Apart from reactive oxygen species mediated by 10.1167/iovs.12-10312 photo-oxidation, hemochromatosis patients may have addi- tional oxidative stress due to iron overload in the retina. In the last decade, several studies have shown the expression and function of iron-regulatory proteins in the retina.11–14 Disrup- tion or loss of iron-regulatory proteins results in AMD-like phenotype in mouse retinas.15,16 Recently, we reported age- From the Departments of 1Biochemistry and Molecular Biology and 2Cellular Biology and Anatomy, Georgia Health Sciences dependent retinal degeneration in Hfe and Hjv knockout 17,18 University, Augusta, Georgia. mice. Interestingly, we found evidence of hypertrophy and Supported by a grant from the National Eye Institute hyperproliferation in retinal pigment epithelial cells (RPE) in (EY019672). both of these mouse models of hemochromatosis. Here we Submitted for publication June 2, 2012; revised October 3 and report that HfeÀ/À RPE cells exhibit several features of tumor November 9, 2012; accepted November 9, 2012. cells. These alterations were also found in HjvÀ/À RPE cells, Disclosure: J.P. Gnana-Prakasam,None;R. Veeranan-Kar- indicating that iron overload is the common underlying cause megam,None;V. Coothankandaswamy,None;S.K. Reddy, for these phenotypic changes. None; P.M. Martin,None;M. Thangaraju,None;S.B. Smith, None; V. Ganapathy,None Cancer cells are dependent on large amounts of micronu- Corresponding author: Jaya P. Gnana-Prakasam, Department of trients like iron for their rapid growth and cell division. There Biochemistry and Molecular Biology, Georgia Health Sciences is evidence that high body iron stores increase risk of cancer in University, Augusta, GA 30912; [email protected]. humans.19 Cancer cells have an uncontrolled capacity for Investigative Ophthalmology & Visual Science, January 2013, Vol. 54, No. 1 Copyright 2013 The Association for Research in Vision and Ophthalmology, Inc. 63 Downloaded from iovs.arvojournals.org on 09/27/2021 64 Gnana-Prakasam et al. IOVS, January 2013, Vol. 54, No. 1 proliferation, migration and invasion. Increase in glucose top of the membrane filter, the inserts were stained with 0.1% crystal uptake and utilization protects cancer cells from starvation. violet, washed, and dried. The crystal violet dye retained on the bottom Tumor progression is not only restricted to the above genetic of the filters (representing the cells that migrated to the underside of modifications, it also involves epigenetic changes. Histone the membrane filter) was extracted with 10% acetic acid. The acetylation mediated by histone acetyltransferases (HATs) absorbance of the extract was measured at 560 nm. Cell invasion induces gene transcription, whereas histone hypoacetylation assay was performed similarly using the QCM cell invasion assay kit. mediated by histone deacetylases (HDACs) is associated with The inserts used in this assay were coated with ECMatrix (Millipore, gene silencing. Altered expression of HDACs has been linked to Billerica, MA), representing the extracellular matrix. Serum-starved tumor development since they affect transcription of genes cells were allowed to invade the matrix for 48 hours in a transwell in that regulate critical cellular functions such as cell prolifera- the presence of serum. After removing the cells on the top of the tion, cell-cycle regulation, and apoptosis.20 Although hemo- membrane filter, the inserts were stained with 0.1% crystal violet, chromatosis patients have been well documented to have an washed, and dried. The crystal violet dye retained on the bottom of the increased risk in developing tumors, the molecular mechanism filters (representing the cells that invaded the matrix to the underside by which iron overload leads to increased predisposition to of the membrane filter) was extracted with 10% acetic acid. The cancer is unknown. In the present study, we found that HfeÀ/À absorbance of the extract was measured at 560 nm. RPE cells have a characteristic tumor phenotype such as increased glucose uptake, enhanced migration, upregulation of Real-Time Polymerase Chain Reaction survivin, and also epigenetic changes that are found in tumor cells. Most of these findings hold true in HjvÀ/À RPE cells as RNA was isolated from wild type and HfeÀ/À or HjvÀ/À primary RPE well. cells in passages three to five, and used for RT-PCR using GeneAmp RT- PCR kit (Applied Biosystems Inc., Carlsbad, CA). Semi-quantitative PCR was performed using Taq polymerase kit (TaKaRa, Shiga, Japan). Real- METHODS time amplifications, using SYBR green detection chemistry, were run in triplicates on 96-well reaction plates. The expression levels of target Animals mRNA was quantified by normalizing with the corresponding internal Breeding pairs of Hfeþ/À mice (C56Bl/6 genetic background) were control (18S RNA). Each PCR experiment was repeated at least three obtained from the Jackson Laboratory (Bar Harbor, ME). Breeding pairs times with similar results. The primers for real-time PCR (Table) were of Hjvþ/À mice (Sv129 genetic background) were provided by Nancy designed based on the sequence information available in GenBank Andrews (Duke University School of Medicine, Durham, NC). (http://www.ncbi.nlm.nih.gov, provided in the public domain by the Genotyping was done to identify wild type, heterozygous, and National Institutes of Health, Bethesda, MD) for mouse cDNAs. homozygous mice in the litters.
Load more

Recommended publications
  • Iron Transport Proteins: Gateways of Cellular and Systemic Iron Homeostasis
    Iron transport proteins: Gateways of cellular and systemic iron homeostasis Mitchell D. Knutson, PhD University of Florida Essential Vocabulary Fe Heme Membrane Transport DMT1 FLVCR Ferroportin HRG1 Mitoferrin Nramp1 ZIP14 Serum Transport Transferrin Transferrin receptor 1 Cytosolic Transport PCBP1, PCBP2 Timeline of identification in mammalian iron transport Year Protein Original Publications 1947 Transferrin Laurell and Ingelman, Acta Chem Scand 1959 Transferrin receptor 1 Jandl et al., J Clin Invest 1997 DMT1 Gunshin et al., Nature; Fleming et al. Nature Genet. 1999 Nramp1 Barton et al., J Leukocyt Biol 2000 Ferroportin Donovan et al., Nature; McKie et al., Cell; Abboud et al. J. Biol Chem 2004 FLVCR Quigley et al., Cell 2006 Mitoferrin Shaw et al., Nature 2006 ZIP14 Liuzzi et al., Proc Natl Acad Sci USA 2008 PCBP1, PCBP2 Shi et al., Science 2013 HRG1 White et al., Cell Metab DMT1 (SLC11A2) • Divalent metal-ion transporter-1 • Former names: Nramp2, DCT1 Fleming et al. Nat Genet, 1997; Gunshin et al., Nature 1997 • Mediates uptake of Fe2+, Mn2+, Cd2+ • H+ coupled transporter (cotransporter, symporter) • Main roles: • intestinal iron absorption Illing et al. JBC, 2012 • iron assimilation by erythroid cells DMT1 (SLC11A2) Yanatori et al. BMC Cell Biology 2010 • 4 different isoforms: 557 – 590 a.a. (hDMT1) Hubert & Hentze, PNAS, 2002 • Function similarly in iron transport • Differ in tissue/subcellular distribution and regulation • Regulated by iron: transcriptionally (via HIF2α) post-transcriptionally (via IRE) IRE = Iron-Responsive Element Enterocyte Lumen DMT1 Fe2+ Fe2+ Portal blood Enterocyte Lumen DMT1 Fe2+ Fe2+ Fe2+ Fe2+ Ferroportin Portal blood Ferroportin (SLC40A1) • Only known mammalian iron exporter Donovan et al., Nature 2000; McKie et al., Cell 2000; Abboud et al.
    [Show full text]
  • Trafficking to the Cell Surface of Amino Acid Transporter SLC6A14
    cells Article Trafficking to the Cell Surface of Amino Acid Transporter SLC6A14 Upregulated in Cancer Is Controlled by Phosphorylation of SEC24C Protein by AKT Kinase Vasylyna Kovalchuk and Katarzyna A. Nał˛ecz* Nencki Institute of Experimental Biology, Polish Academy of Sciences, 3 Pasteur Street, PL-02-093 Warsaw, Poland; [email protected] * Correspondence: [email protected]; Tel.: +48-225892303 Abstract: Cancer cells need a constant supply of nutrients. SLC6A14, an amino acid transporter B0,+ (ATB0,+) that is upregulated in many cancers, transports all but acidic amino acids. In its exit from the endoplasmic reticulum (ER), it is recognized by the SEC24C subunit of coatomer II (COPII) for further vesicular trafficking to the plasma membrane. SEC24C has previously been shown to be phosphorylated by protein kinase B/AKT, which is hyper-activated in cancer; therefore, we analyzed the influence of AKT on SLC6A14 trafficking to the cell surface. Studies on overexpressed and en- dogenous transporters in the breast cancer cell line MCF-7 showed that AKT inhibition with MK-2206 correlated with a transient increase of the transporter in the plasma membrane, not resulting from the inhibition of ER-associated protein degradation. Two-dimensional electrophoresis demonstrated the decreased phosphorylation of SLC6A14 and SEC24C upon AKT inhibition. A proximity ligation assay confirmed this conclusion: AKT inhibition is correlated with decreased SLC6A14 phosphothreonine Citation: Kovalchuk, V.; Nał˛ecz,K.A. Trafficking to the Cell Surface of and SEC24C phosphoserine. Augmented levels of SLC6A14 in plasma membrane led to increased Amino Acid Transporter SLC6A14 leucine transport. These results show that the inactivation of AKT can rescue amino acid delivery Upregulated in Cancer Is Controlled through SLC6A14 trafficking to the cell surface, supporting cancer cell survival.
    [Show full text]
  • A Computational Approach for Defining a Signature of Β-Cell Golgi Stress in Diabetes Mellitus
    Page 1 of 781 Diabetes A Computational Approach for Defining a Signature of β-Cell Golgi Stress in Diabetes Mellitus Robert N. Bone1,6,7, Olufunmilola Oyebamiji2, Sayali Talware2, Sharmila Selvaraj2, Preethi Krishnan3,6, Farooq Syed1,6,7, Huanmei Wu2, Carmella Evans-Molina 1,3,4,5,6,7,8* Departments of 1Pediatrics, 3Medicine, 4Anatomy, Cell Biology & Physiology, 5Biochemistry & Molecular Biology, the 6Center for Diabetes & Metabolic Diseases, and the 7Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202; 2Department of BioHealth Informatics, Indiana University-Purdue University Indianapolis, Indianapolis, IN, 46202; 8Roudebush VA Medical Center, Indianapolis, IN 46202. *Corresponding Author(s): Carmella Evans-Molina, MD, PhD ([email protected]) Indiana University School of Medicine, 635 Barnhill Drive, MS 2031A, Indianapolis, IN 46202, Telephone: (317) 274-4145, Fax (317) 274-4107 Running Title: Golgi Stress Response in Diabetes Word Count: 4358 Number of Figures: 6 Keywords: Golgi apparatus stress, Islets, β cell, Type 1 diabetes, Type 2 diabetes 1 Diabetes Publish Ahead of Print, published online August 20, 2020 Diabetes Page 2 of 781 ABSTRACT The Golgi apparatus (GA) is an important site of insulin processing and granule maturation, but whether GA organelle dysfunction and GA stress are present in the diabetic β-cell has not been tested. We utilized an informatics-based approach to develop a transcriptional signature of β-cell GA stress using existing RNA sequencing and microarray datasets generated using human islets from donors with diabetes and islets where type 1(T1D) and type 2 diabetes (T2D) had been modeled ex vivo. To narrow our results to GA-specific genes, we applied a filter set of 1,030 genes accepted as GA associated.
    [Show full text]
  • Ferredoxin Reductase Is Critical for P53-Dependent Tumor Suppression Via Iron Regulatory Protein 2
    Downloaded from genesdev.cshlp.org on October 11, 2021 - Published by Cold Spring Harbor Laboratory Press Ferredoxin reductase is critical for p53- dependent tumor suppression via iron regulatory protein 2 Yanhong Zhang,1,9 Yingjuan Qian,1,2,9 Jin Zhang,1 Wensheng Yan,1 Yong-Sam Jung,1,2 Mingyi Chen,3 Eric Huang,4 Kent Lloyd,5 Yuyou Duan,6 Jian Wang,7 Gang Liu,8 and Xinbin Chen1 1Comparative Oncology Laboratory, Schools of Veterinary Medicine and Medicine, University of California at Davis, Davis, California 95616, USA; 2College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210014, China; 3Department of Pathology, University of Texas Southwestern Medical Center, Dallas, Texas 75390, USA; 4Department of Pathology, School of Medicine, University of California at Davis Health, Sacramento, California 95817, USA; 5Department of Surgery, School of Medicine, University of California at Davis Health, Sacramento, California 95817, USA; 6Department of Dermatology and Internal Medicine, University of California at Davis Health, Sacramento, California 95616, USA; 7Department of Pathology, School of Medicine, Wayne State University, Detroit, Michigan 48201 USA; 8Department of Medicine, School of Medicine, University of Alabama at Birmingham, Birmingham, Alabama 35294, USA Ferredoxin reductase (FDXR), a target of p53, modulates p53-dependent apoptosis and is necessary for steroido- genesis and biogenesis of iron–sulfur clusters. To determine the biological function of FDXR, we generated a Fdxr- deficient mouse model and found that loss of Fdxr led to embryonic lethality potentially due to iron overload in developing embryos. Interestingly, mice heterozygous in Fdxr had a short life span and were prone to spontaneous tumors and liver abnormalities, including steatosis, hepatitis, and hepatocellular carcinoma.
    [Show full text]
  • Supplementary Table S4. FGA Co-Expressed Gene List in LUAD
    Supplementary Table S4. FGA co-expressed gene list in LUAD tumors Symbol R Locus Description FGG 0.919 4q28 fibrinogen gamma chain FGL1 0.635 8p22 fibrinogen-like 1 SLC7A2 0.536 8p22 solute carrier family 7 (cationic amino acid transporter, y+ system), member 2 DUSP4 0.521 8p12-p11 dual specificity phosphatase 4 HAL 0.51 12q22-q24.1histidine ammonia-lyase PDE4D 0.499 5q12 phosphodiesterase 4D, cAMP-specific FURIN 0.497 15q26.1 furin (paired basic amino acid cleaving enzyme) CPS1 0.49 2q35 carbamoyl-phosphate synthase 1, mitochondrial TESC 0.478 12q24.22 tescalcin INHA 0.465 2q35 inhibin, alpha S100P 0.461 4p16 S100 calcium binding protein P VPS37A 0.447 8p22 vacuolar protein sorting 37 homolog A (S. cerevisiae) SLC16A14 0.447 2q36.3 solute carrier family 16, member 14 PPARGC1A 0.443 4p15.1 peroxisome proliferator-activated receptor gamma, coactivator 1 alpha SIK1 0.435 21q22.3 salt-inducible kinase 1 IRS2 0.434 13q34 insulin receptor substrate 2 RND1 0.433 12q12 Rho family GTPase 1 HGD 0.433 3q13.33 homogentisate 1,2-dioxygenase PTP4A1 0.432 6q12 protein tyrosine phosphatase type IVA, member 1 C8orf4 0.428 8p11.2 chromosome 8 open reading frame 4 DDC 0.427 7p12.2 dopa decarboxylase (aromatic L-amino acid decarboxylase) TACC2 0.427 10q26 transforming, acidic coiled-coil containing protein 2 MUC13 0.422 3q21.2 mucin 13, cell surface associated C5 0.412 9q33-q34 complement component 5 NR4A2 0.412 2q22-q23 nuclear receptor subfamily 4, group A, member 2 EYS 0.411 6q12 eyes shut homolog (Drosophila) GPX2 0.406 14q24.1 glutathione peroxidase
    [Show full text]
  • Essential Trace Elements in Human Health: a Physician's View
    Margarita G. Skalnaya, Anatoly V. Skalny ESSENTIAL TRACE ELEMENTS IN HUMAN HEALTH: A PHYSICIAN'S VIEW Reviewers: Philippe Collery, M.D., Ph.D. Ivan V. Radysh, M.D., Ph.D., D.Sc. Tomsk Publishing House of Tomsk State University 2018 2 Essential trace elements in human health UDK 612:577.1 LBC 52.57 S66 Skalnaya Margarita G., Skalny Anatoly V. S66 Essential trace elements in human health: a physician's view. – Tomsk : Publishing House of Tomsk State University, 2018. – 224 p. ISBN 978-5-94621-683-8 Disturbances in trace element homeostasis may result in the development of pathologic states and diseases. The most characteristic patterns of a modern human being are deficiency of essential and excess of toxic trace elements. Such a deficiency frequently occurs due to insufficient trace element content in diets or increased requirements of an organism. All these changes of trace element homeostasis form an individual trace element portrait of a person. Consequently, impaired balance of every trace element should be analyzed in the view of other patterns of trace element portrait. Only personalized approach to diagnosis can meet these requirements and result in successful treatment. Effective management and timely diagnosis of trace element deficiency and toxicity may occur only in the case of adequate assessment of trace element status of every individual based on recent data on trace element metabolism. Therefore, the most recent basic data on participation of essential trace elements in physiological processes, metabolism, routes and volumes of entering to the body, relation to various diseases, medical applications with a special focus on iron (Fe), copper (Cu), manganese (Mn), zinc (Zn), selenium (Se), iodine (I), cobalt (Co), chromium, and molybdenum (Mo) are reviewed.
    [Show full text]
  • Analysis of Dynamic Molecular Networks for Pancreatic Ductal
    Pan et al. Cancer Cell Int (2018) 18:214 https://doi.org/10.1186/s12935-018-0718-5 Cancer Cell International PRIMARY RESEARCH Open Access Analysis of dynamic molecular networks for pancreatic ductal adenocarcinoma progression Zongfu Pan1†, Lu Li2†, Qilu Fang1, Yiwen Zhang1, Xiaoping Hu1, Yangyang Qian3 and Ping Huang1* Abstract Background: Pancreatic ductal adenocarcinoma (PDAC) is one of the deadliest solid tumors. The rapid progression of PDAC results in an advanced stage of patients when diagnosed. However, the dynamic molecular mechanism underlying PDAC progression remains far from clear. Methods: The microarray GSE62165 containing PDAC staging samples was obtained from Gene Expression Omnibus and the diferentially expressed genes (DEGs) between normal tissue and PDAC of diferent stages were profled using R software, respectively. The software program Short Time-series Expression Miner was applied to cluster, compare, and visualize gene expression diferences between PDAC stages. Then, function annotation and pathway enrichment of DEGs were conducted by Database for Annotation Visualization and Integrated Discovery. Further, the Cytoscape plugin DyNetViewer was applied to construct the dynamic protein–protein interaction networks and to analyze dif- ferent topological variation of nodes and clusters over time. The phosphosite markers of stage-specifc protein kinases were predicted by PhosphoSitePlus database. Moreover, survival analysis of candidate genes and pathways was per- formed by Kaplan–Meier plotter. Finally, candidate genes were validated by immunohistochemistry in PDAC tissues. Results: Compared with normal tissues, the total DEGs number for each PDAC stage were 994 (stage I), 967 (stage IIa), 965 (stage IIb), 1027 (stage III), 925 (stage IV), respectively. The stage-course gene expression analysis showed that 30 distinct expressional models were clustered.
    [Show full text]
  • Microarray Analysis Reveals the Inhibition of Intestinal Expression Of
    www.nature.com/scientificreports OPEN Microarray analysis reveals the inhibition of intestinal expression of nutrient transporters in piglets infected with porcine epidemic diarrhea virus Junmei Zhang1,3, Di Zhao1,3, Dan Yi1,3, Mengjun Wu1, Hongbo Chen1, Tao Wu1, Jia Zhou1, Peng Li1, Yongqing Hou1* & Guoyao Wu2 Porcine epidemic diarrhea virus (PEDV) infection can induce intestinal dysfunction, resulting in severe diarrhea and even death, but the mode of action underlying these viral efects remains unclear. This study determined the efects of PEDV infection on intestinal absorption and the expression of genes for nutrient transporters via biochemical tests and microarray analysis. Sixteen 7-day-old healthy piglets fed a milk replacer were randomly allocated to one of two groups. After 5-day adaption, piglets (n = 8/ group) were orally administrated with either sterile saline or PEDV (the strain from Yunnan province) 4.5 at 10 TCID50 (50% tissue culture infectious dose) per pig. All pigs were orally infused D-xylose (0.1 g/ kg BW) on day 5 post PEDV or saline administration. One hour later, jugular vein blood samples as well as intestinal samples were collected for further analysis. In comparison with the control group, PEDV infection increased diarrhea incidence, blood diamine oxidase activity, and iFABP level, while reducing growth and plasma D-xylose concentration in piglets. Moreover, PEDV infection altered plasma and jejunal amino acid profles, and decreased the expression of aquaporins and amino acid transporters (L-type amino acid
    [Show full text]
  • Comparative Transcriptome Profiling of Selected Osmotic Regulatory Proteins in the Gill During Seawater Acclimation of Chum Salm
    www.nature.com/scientificreports OPEN Comparative transcriptome profling of selected osmotic regulatory proteins in the gill during seawater acclimation of chum salmon (Oncorhynchus keta) fry Sang Yoon Lee 1, Hwa Jin Lee2 & Yi Kyung Kim1,2* Salmonid fshes, chum salmon (Oncorhynchus keta) have the developed adaptive strategy to withstand wide salinity changes from the early life stage. This study investigated gene expression patterns of cell membrane proteins in the gill of chum salmon fry on the transcriptome level by tracking the salinity acclimation of the fsh in changing environments ranging from freshwater (0 ppt) to brackish water (17.5 ppt) to seawater (35 ppt). Using GO analysis of DEGs, the known osmoregulatory genes and their functional groups such as ion transport, transmembrane transporter activity and metal ion binding were identifed. The expression patterns of membrane protein genes, including pump-mediated protein (NKA, CFTR), carrier-mediated protein (NKCC, NHE3) and channel-mediated protein (AQP) were similar to those of other salmonid fshes in the smolt or adult stages. Based on the protein-protein interaction analysis between transmembrane proteins and other related genes, we identifed osmotic-related genes expressed with salinity changes and analyzed their expression patterns. The fndings of this study may facilitate the disentangling of the genetic basis of chum salmon and better able an understanding of the osmophysiology of the species. Salinity is one of the critical factors limiting the distribution patterns of all aquatic organisms1–4. Salmonid fshes display diverse life-history traits; anadromous individuals that mature in the river from hatching through to juveniles acquire the capacity to tolerate salinity associated with parr–smolt transformation and undergo ocean migrations before returning to rivers for spawning, whereas landlocked types spend their entire life within fresh- water5,6.
    [Show full text]
  • Sudden Sensorineural Hearing Loss and Polymorphisms in Iron Homeostasis Genes: New Insights from a Case-Control Study
    Hindawi Publishing Corporation BioMed Research International Volume 2015, Article ID 834736, 10 pages http://dx.doi.org/10.1155/2015/834736 Research Article Sudden Sensorineural Hearing Loss and Polymorphisms in Iron Homeostasis Genes: New Insights from a Case-Control Study Alessandro Castiglione,1 Andrea Ciorba,2 Claudia Aimoni,2 Elisa Orioli,3 Giulia Zeri,3 Marco Vigliano,3 and Donato Gemmati3 1 Department of Neurosciences-Complex Operative Unit of Otorhinolaryngology and Otosurgery, University Hospital of Padua, Via Giustiniani 2, 35128 Padua, Italy 2ENT & Audiology Department, University Hospital of Ferrara, Via Aldo Moro 8, 44124 Cona, Ferrara, Italy 3Centre for Haemostasis & Thrombosis, Haematology Section, Department of Medical Sciences, University of Ferrara, 44100 Ferrara, Italy Correspondence should be addressed to Alessandro Castiglione; [email protected] Received 15 September 2014; Revised 15 December 2014; Accepted 6 January 2015 Academic Editor: Song Liu Copyright © 2015 Alessandro Castiglione et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Background. Even if various pathophysiological events have been proposed as explanations, the putative cause of sudden hearing loss remains unclear. Objectives. To investigate and to reveal associations (if any) between the main iron-related gene variants and idiopathic sudden sensorineural hearing loss.
    [Show full text]
  • A Short Review of Iron Metabolism and Pathophysiology of Iron Disorders
    medicines Review A Short Review of Iron Metabolism and Pathophysiology of Iron Disorders Andronicos Yiannikourides 1 and Gladys O. Latunde-Dada 2,* 1 Faculty of Life Sciences and Medicine, Henriette Raphael House Guy’s Campus King’s College London, London SE1 1UL, UK 2 Department of Nutritional Sciences, School of Life Course Sciences, King’s College London, Franklin-Wilkins-Building, 150 Stamford Street, London SE1 9NH, UK * Correspondence: [email protected] Received: 30 June 2019; Accepted: 2 August 2019; Published: 5 August 2019 Abstract: Iron is a vital trace element for humans, as it plays a crucial role in oxygen transport, oxidative metabolism, cellular proliferation, and many catalytic reactions. To be beneficial, the amount of iron in the human body needs to be maintained within the ideal range. Iron metabolism is one of the most complex processes involving many organs and tissues, the interaction of which is critical for iron homeostasis. No active mechanism for iron excretion exists. Therefore, the amount of iron absorbed by the intestine is tightly controlled to balance the daily losses. The bone marrow is the prime iron consumer in the body, being the site for erythropoiesis, while the reticuloendothelial system is responsible for iron recycling through erythrocyte phagocytosis. The liver has important synthetic, storing, and regulatory functions in iron homeostasis. Among the numerous proteins involved in iron metabolism, hepcidin is a liver-derived peptide hormone, which is the master regulator of iron metabolism. This hormone acts in many target tissues and regulates systemic iron levels through a negative feedback mechanism. Hepcidin synthesis is controlled by several factors such as iron levels, anaemia, infection, inflammation, and erythropoietic activity.
    [Show full text]
  • Investigating the Role of Amino Acid Transporter SLC6A14 As a Modifier of Pseudomonas Aeruginosa Lung Infection in Cystic Fibrosis
    Investigating the Role of Amino Acid Transporter SLC6A14 as a Modifier of Pseudomonas aeruginosa Lung Infection in Cystic Fibrosis by Michelle Di Paola A thesis submitted in conformity with the requirements for the degree of Master of Science Biochemistry University of Toronto © Copyright by Michelle Di Paola (2016) Investigating the Role of Amino Acid Transporter SLC6A14 as a Modifier of Pseudomonas aeruginosa Lung Infection in Cystic Fibrosis Michelle Di Paola Master of Science Biochemistry University of Toronto 2016 Abstract The most common disease causing mutation in CF is F508del, however there is considerable variability in the clinical phenotype of patients homozygous for this mutation. To address this variation, genetic modifiers are studied. Li et al found that SNPs in the putative promoter region of SLC6A14 were significantly associated with severity and age of first Pseudomonas aeruginosa lung infection in CF. Experiments were aimed at studying if SLC6A14-mediated amino acid uptake is regulated by bacterial pathogens and in turn regulates bacterial growth. Arginine uptake from the ASL of non-CF and CF primary bronchial cultures was measured, after treatment with purified flagellin from Pseudomonas. This treatment resulted in a 26.4% (n=5; p<0.01) and 16.6% (n=6; p<0.0204) increase in arginine uptake, in non-CF and CF cultures respectively. Currently, the impact of SLC6A14 function on Pseudomonas growth in co-culture is being investigated, to better understand the role which SLC6A14 plays in modulating infection. ii Acknowledgments I first need to extend my deepest gratitude to my supervisor, Dr. Christine Bear, for providing me with endless support and guidance.
    [Show full text]