DOCSLIB.ORG

History of Iga Nephropathy Mouse Models

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
History of Iga Nephropathy Mouse Models Journal of Clinical Medicine Review History of IgA Nephropathy Mouse Models Batoul Wehbi 1, Virginie Pascal 1, Lina Zawil 1, Michel Cogné 2 and Jean-Claude Aldigier 1,* 1 Immunology Department, UMR CNRS 7276 INSERM 1262, Limoges University, 87032 Limoges, France; [email protected] (B.W.); [email protected] (V.P.); [email protected] (L.Z.) 2 Immunology Department, EFS Bretagne, INSERM 1236, Rennes 1 University, 35000 Rennes, France; [email protected] * Correspondence: [email protected] Abstract: IgA nephropathy (IgAN) is the most common primary glomerulonephritis in the world. It was first described in 1968 by Jean Berger and Nicole Hinglais as the presence of intercapillary deposits of IgA. Despite this simple description, patients with IgAN may present very broad clinical features ranging from the isolated presence of IgA in the mesangium without clinical or biological manifestations to rapidly progressive kidney failure. These features are associated with a variety of histological lesions, from the discrete thickening of the mesangial matrix to diffuse cell proliferation. Immunofluorescence on IgAN kidney specimens shows the isolated presence of IgA or its inconsistent association with IgG and complement components. This clinical heterogeneity of IgAN clearly echoes its complex and multifactorial pathophysiology in humans, inviting further analyses of its various aspects through the use of experimental models. Small-animal models of IgAN provide the most pertinent strategies for studying the multifactorial aspects of IgAN pathogenesis and progression. Although only primates have the IgA1 subclass, several murine models have been developed in which various aspects of immune responses are deregulated and which are useful in the understanding of IgAN physiopathology as well as in the assessment of IgAN therapeutic approaches. In this Citation: Wehbi, B.; Pascal, V.; manuscript, we review all murine IgAN models developed since 1968 and discuss their remarkable Zawil, L.; Cogné, M.; Aldigier, J.-C. History of IgA Nephropathy Mouse contribution to understanding the disease. Models. J. Clin. Med. 2021, 10, 3142. https://doi.org/10.3390/ Keywords: IgA nephropathy; IgA; kidney mesangium; mouse model jcm10143142 Academic Editors: Hitoshi Suzuki and Jan Novak 1. IgAN Epidemiology The true prevalence of IgAN cannot be exactly determined. IgA deposits on the kidney Received: 6 May 2021 are indeed frequent in asymptomatic patients and were reported in 16.1% of a population of Accepted: 9 July 2021 kidney donors in Japan [1]. However, the overt disease can evolve to a rapidly progressive Published: 16 July 2021 glomerulopathy (Figure1). Since the diagnosis of IgAN requires a kidney biopsy, available data only refer to cases recorded after this procedure. The prevalence of IgAN varies widely Publisher’s Note: MDPI stays neutral across different geographic regions and ethnic groups [2]: IgAN is most prevalent in East with regard to jurisdictional claims in Asian people (with more than 40% of biopsy cases in Japan), followed by Caucasians and published maps and institutional affil- African individuals (respectively with about 25% and less than 5% of biopsy cases) [3]. iations. This disparity in population distribution can be attributed to the different health screening policies as well as several genetic and environmental factors. Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/). J. Clin. Med. 2021, 10, 3142. https://doi.org/10.3390/jcm10143142 https://www.mdpi.com/journal/jcm J. Clin. Med. 2021, 10, 3142 2 of 11 J. Clin. Med. 2021, 10, x FOR PEER REVIEW 2 of 11 FigureFigure 1.1. IgANIgAN represent represent the the emergent emergent part part of of the the IgA IgA deposition deposition iceberg iceberg with with a heterogeneity a heterogeneity un- underscoringderscoring the the multifactorial multifactorial pathogenesis pathogenesis of ofthe the disease. disease. 2.2. IgAN:IgAN: ClinicalClinical andand HistopathologicalHistopathological PresentationPresentation ThereThere isis significantsignificant heterogeneityheterogeneity inin thethe clinicalclinical manifestationsmanifestations ofof IgANIgAN sincesince eveneven healthyhealthy subjectssubjects withwith nono clinicalclinical signssigns ofof IgANIgAN maymay havehave mesangialmesangial deposits.deposits. ByBy contrast,contrast, forfor thosethose patientspatients developingdeveloping glomerulopathyglomerulopathy withwith proteinuriaproteinuria and/orand/or hematuria,hematuria, one-one- thirdthird ofof thethe casescases willwill progressprogress toto endend stagestage renalrenal diseasedisease (ESRD)(ESRD) [[4].4]. HighHigh bloodblood pressurepressure maymay alsoalso be an indicative indicative sign sign of of IgAN; IgAN; it itis isalways always present present as assoon soon as askidney kidney failure failure be- begins.gins. In Inpatients patients presenting presenting such such features, features, renal renal biopsy biopsy is the is the only only method method to confirm to confirm di- diagnosis.agnosis. The The anatomo anatomo-pathological-pathological study study of ofrenal renal biopsy biopsy specimens specimens reveals reveals mesangial mesangial le- lesions:sions: mesangial mesangial matrix matrix expansion expansion and and hypercellularity sometimes associated withwith aa mesangialmesangial cellcell proliferationproliferation withwith possiblepossible endo-endo- oror extra-capillaryextra-capillary proliferationproliferation oror eveneven significantsignificant glomerularglomerular inflammationinflammation andand crescentcrescent formation.formation. OtherOther lesionslesions cancan alsoalso bebe identified:identified: segmentalsegmental sclerosis,sclerosis, tubulartubular atrophy,atrophy, andand interstitialinterstitial fibrosisfibrosis [5[5].]. TheThe analysisanalysis ofof thesethese differentdifferent lesionslesions makesmakes itit possiblepossible toto establishestablish thethe OxfordOxford predictivepredictive scorescore forfor diseasedisease progressionprogression [[6].6]. Immunofluorescence Immunofluorescence on on biopsies biopsies reveals reveals the the presence presence of ofmore more or orless less diffuse diffuse granular granular or or filamentous filamentous IgA IgA deposits deposits in in the the form form of “dead“dead trees”trees” oror “nail“nail strokes”strokes”[ [5].5]. TheThe presencepresence ofof lambdalambda andand kappakappa lightlight chainchain depositsdeposits hashas beenbeen reportedreported [[7].7]. IgAIgA deposits deposits most most often often co-localize co-localize with with C3, C3, sometimes sometimes with with IgG IgG and and IgM IgM [3]. Moreover,[3]. Moreover, the presencethe presence of deposits of deposits at the at the level level of singleof single glomeruli glomeruli is is sufficient sufficient to to make make the the diagnosis diagnosis [[5]5].. 3. Enigmatic Functions of IgA 3. Enigmatic Functions of IgA IgA antibodies (Ab) are exceptionally abundant at mucosal surfaces; more than 80% IgA antibodies (Ab) are exceptionally abundant at mucosal surfaces; more than 80% of mammalian Ab secreting plasma cells reside in the gut and secrete IgA. Notably, with of mammalian Ab secreting plasma cells reside in the gut and secrete IgA. Notably, with the exception of germ-free animals or germ-free mucosal tissues, these antibodies arise the exception of germ-free animals or germ-free mucosal tissues, these antibodies arise prominently during homeostasis in the absence of overt inflammation and immunization. Consistently,prominently during the influence homeo ofstasis super in the antigens absence expressed of overt inflammation by bacteria present and immunization. early in the intestinalConsistently, flora the has influence recently of been super demonstrated antigens expressed [8]. Despite by bacteria its abundance, present early the in in vivo the specificityintestinal flora and dualhas recently functions been of IgAdemonstrated still remain [8]. somehow Despite enigmatic,its abundance, although the in these vivo antibodiesspecificity and appear dual as functions major players of IgA instill the remain functional somehow balance enigmatic, opposing although tolerance these and an- inflammationtibodies appear in as the major context players of mucosal in the functional immunity balance and as opposing important tolerance organizers and inflam- of the microbiotamation in the composition context of mucosal in healthy immunity subjects and [9]. as At important the gut-associated organizers lymphoidof the microbiota tissue (GALT)compositio level,n in there healthy are two subjects distinct [9]. aspects At the of gut humoral-associated immunity: lymphoid first, tissue the mechanisms (GALT) level, of J. Clin. Med. 2021, 10, 3142 3 of 11 humoral homeostasis against commensal bacteria seem to involve the so-called “natural IgA”. Their repertoires are almost germline with slightly mature variable regions [10]. These IgA share an interspecies reactivity because they are often polyreactive and partly produced by B-lymphocytes in a T-independent context. Second, during the local immune response induced by pathogens, protective high affinity IgA are produced in the mucosa-associated lymphoid tissue (MALT) germinal centers. This T-dependent response is similar to that of a general response establishing specific immunity as effective as a response
Load more

Recommended publications
  • Supplemental Figure 1. Vimentin
    Double mutant specific genes Transcript gene_assignment Gene Symbol RefSeq FDR Fold- FDR Fold- FDR Fold- ID (single vs. Change (double Change (double Change wt) (single vs. wt) (double vs. single) (double vs. wt) vs. wt) vs. single) 10485013 BC085239 // 1110051M20Rik // RIKEN cDNA 1110051M20 gene // 2 E1 // 228356 /// NM 1110051M20Ri BC085239 0.164013 -1.38517 0.0345128 -2.24228 0.154535 -1.61877 k 10358717 NM_197990 // 1700025G04Rik // RIKEN cDNA 1700025G04 gene // 1 G2 // 69399 /// BC 1700025G04Rik NM_197990 0.142593 -1.37878 0.0212926 -3.13385 0.093068 -2.27291 10358713 NM_197990 // 1700025G04Rik // RIKEN cDNA 1700025G04 gene // 1 G2 // 69399 1700025G04Rik NM_197990 0.0655213 -1.71563 0.0222468 -2.32498 0.166843 -1.35517 10481312 NM_027283 // 1700026L06Rik // RIKEN cDNA 1700026L06 gene // 2 A3 // 69987 /// EN 1700026L06Rik NM_027283 0.0503754 -1.46385 0.0140999 -2.19537 0.0825609 -1.49972 10351465 BC150846 // 1700084C01Rik // RIKEN cDNA 1700084C01 gene // 1 H3 // 78465 /// NM_ 1700084C01Rik BC150846 0.107391 -1.5916 0.0385418 -2.05801 0.295457 -1.29305 10569654 AK007416 // 1810010D01Rik // RIKEN cDNA 1810010D01 gene // 7 F5 // 381935 /// XR 1810010D01Rik AK007416 0.145576 1.69432 0.0476957 2.51662 0.288571 1.48533 10508883 NM_001083916 // 1810019J16Rik // RIKEN cDNA 1810019J16 gene // 4 D2.3 // 69073 / 1810019J16Rik NM_001083916 0.0533206 1.57139 0.0145433 2.56417 0.0836674 1.63179 10585282 ENSMUST00000050829 // 2010007H06Rik // RIKEN cDNA 2010007H06 gene // --- // 6984 2010007H06Rik ENSMUST00000050829 0.129914 -1.71998 0.0434862 -2.51672
    [Show full text]
  • Appendix 2. Significantly Differentially Regulated Genes in Term Compared with Second Trimester Amniotic Fluid Supernatant
    Appendix 2. Significantly Differentially Regulated Genes in Term Compared With Second Trimester Amniotic Fluid Supernatant Fold Change in term vs second trimester Amniotic Affymetrix Duplicate Fluid Probe ID probes Symbol Entrez Gene Name 1019.9 217059_at D MUC7 mucin 7, secreted 424.5 211735_x_at D SFTPC surfactant protein C 416.2 206835_at STATH statherin 363.4 214387_x_at D SFTPC surfactant protein C 295.5 205982_x_at D SFTPC surfactant protein C 288.7 1553454_at RPTN repetin solute carrier family 34 (sodium 251.3 204124_at SLC34A2 phosphate), member 2 238.9 206786_at HTN3 histatin 3 161.5 220191_at GKN1 gastrokine 1 152.7 223678_s_at D SFTPA2 surfactant protein A2 130.9 207430_s_at D MSMB microseminoprotein, beta- 99.0 214199_at SFTPD surfactant protein D major histocompatibility complex, class II, 96.5 210982_s_at D HLA-DRA DR alpha 96.5 221133_s_at D CLDN18 claudin 18 94.4 238222_at GKN2 gastrokine 2 93.7 1557961_s_at D LOC100127983 uncharacterized LOC100127983 93.1 229584_at LRRK2 leucine-rich repeat kinase 2 HOXD cluster antisense RNA 1 (non- 88.6 242042_s_at D HOXD-AS1 protein coding) 86.0 205569_at LAMP3 lysosomal-associated membrane protein 3 85.4 232698_at BPIFB2 BPI fold containing family B, member 2 84.4 205979_at SCGB2A1 secretoglobin, family 2A, member 1 84.3 230469_at RTKN2 rhotekin 2 82.2 204130_at HSD11B2 hydroxysteroid (11-beta) dehydrogenase 2 81.9 222242_s_at KLK5 kallikrein-related peptidase 5 77.0 237281_at AKAP14 A kinase (PRKA) anchor protein 14 76.7 1553602_at MUCL1 mucin-like 1 76.3 216359_at D MUC7 mucin 7,
    [Show full text]
  • MALE Protein Name Accession Number Molecular Weight CP1 CP2 H1 H2 PDAC1 PDAC2 CP Mean H Mean PDAC Mean T-Test PDAC Vs. H T-Test
    MALE t-test t-test Accession Molecular H PDAC PDAC vs. PDAC vs. Protein Name Number Weight CP1 CP2 H1 H2 PDAC1 PDAC2 CP Mean Mean Mean H CP PDAC/H PDAC/CP - 22 kDa protein IPI00219910 22 kDa 7 5 4 8 1 0 6 6 1 0.1126 0.0456 0.1 0.1 - Cold agglutinin FS-1 L-chain (Fragment) IPI00827773 12 kDa 32 39 34 26 53 57 36 30 55 0.0309 0.0388 1.8 1.5 - HRV Fab 027-VL (Fragment) IPI00827643 12 kDa 4 6 0 0 0 0 5 0 0 - 0.0574 - 0.0 - REV25-2 (Fragment) IPI00816794 15 kDa 8 12 5 7 8 9 10 6 8 0.2225 0.3844 1.3 0.8 A1BG Alpha-1B-glycoprotein precursor IPI00022895 54 kDa 115 109 106 112 111 100 112 109 105 0.6497 0.4138 1.0 0.9 A2M Alpha-2-macroglobulin precursor IPI00478003 163 kDa 62 63 86 72 14 18 63 79 16 0.0120 0.0019 0.2 0.3 ABCB1 Multidrug resistance protein 1 IPI00027481 141 kDa 41 46 23 26 52 64 43 25 58 0.0355 0.1660 2.4 1.3 ABHD14B Isoform 1 of Abhydrolase domain-containing proteinIPI00063827 14B 22 kDa 19 15 19 17 15 9 17 18 12 0.2502 0.3306 0.7 0.7 ABP1 Isoform 1 of Amiloride-sensitive amine oxidase [copper-containing]IPI00020982 precursor85 kDa 1 5 8 8 0 0 3 8 0 0.0001 0.2445 0.0 0.0 ACAN aggrecan isoform 2 precursor IPI00027377 250 kDa 38 30 17 28 34 24 34 22 29 0.4877 0.5109 1.3 0.8 ACE Isoform Somatic-1 of Angiotensin-converting enzyme, somaticIPI00437751 isoform precursor150 kDa 48 34 67 56 28 38 41 61 33 0.0600 0.4301 0.5 0.8 ACE2 Isoform 1 of Angiotensin-converting enzyme 2 precursorIPI00465187 92 kDa 11 16 20 30 4 5 13 25 5 0.0557 0.0847 0.2 0.4 ACO1 Cytoplasmic aconitate hydratase IPI00008485 98 kDa 2 2 0 0 0 0 2 0 0 - 0.0081 - 0.0
    [Show full text]
  • Urinary Proteomics for the Early Diagnosis of Diabetic Nephropathy in Taiwanese Patients Authors
    Urinary Proteomics for the Early Diagnosis of Diabetic Nephropathy in Taiwanese Patients Authors: Wen-Ling Liao1,2, Chiz-Tzung Chang3,4, Ching-Chu Chen5,6, Wen-Jane Lee7,8, Shih-Yi Lin3,4, Hsin-Yi Liao9, Chia-Ming Wu10, Ya-Wen Chang10, Chao-Jung Chen1,9,+,*, Fuu-Jen Tsai6,10,11,+,* 1 Graduate Institute of Integrated Medicine, China Medical University, Taichung, 404, Taiwan 2 Center for Personalized Medicine, China Medical University Hospital, Taichung, 404, Taiwan 3 Division of Nephrology and Kidney Institute, Department of Internal Medicine, China Medical University Hospital, Taichung, 404, Taiwan 4 Institute of Clinical Medical Science, China Medical University College of Medicine, Taichung, 404, Taiwan 5 Division of Endocrinology and Metabolism, Department of Medicine, China Medical University Hospital, Taichung, 404, Taiwan 6 School of Chinese Medicine, China Medical University, Taichung, 404, Taiwan 7 Department of Medical Research, Taichung Veterans General Hospital, Taichung, 404, Taiwan 8 Department of Social Work, Tunghai University, Taichung, 404, Taiwan 9 Proteomics Core Laboratory, Department of Medical Research, China Medical University Hospital, Taichung, 404, Taiwan 10 Human Genetic Center, Department of Medical Research, China Medical University Hospital, China Medical University, Taichung, 404, Taiwan 11 Department of Health and Nutrition Biotechnology, Asia University, Taichung, 404, Taiwan + Fuu-Jen Tsai and Chao-Jung Chen contributed equally to this work. Correspondence: Fuu-Jen Tsai, MD, PhD and Chao-Jung Chen, PhD FJ Tsai: Genetic Center, China Medical University Hospital, No.2 Yuh-Der Road, 404 Taichung, Taiwan; Telephone: 886-4-22062121 Ext. 2041; Fax: 886-4-22033295; E-mail: [email protected] CJ Chen: Graduate Institute of Integrated Medicine, China Medical University, No.91, Hsueh-Shih Road, 404, Taichung, Taiwan; Telephone: 886-4-22053366 Ext.
    [Show full text]
  • Supplementary Table 1
    Supplementary Table 1. 492 genes are unique to 0 h post-heat timepoint. The name, p-value, fold change, location and family of each gene are indicated. Genes were filtered for an absolute value log2 ration 1.5 and a significance value of p ≤ 0.05. Symbol p-value Log Gene Name Location Family Ratio ABCA13 1.87E-02 3.292 ATP-binding cassette, sub-family unknown transporter A (ABC1), member 13 ABCB1 1.93E-02 −1.819 ATP-binding cassette, sub-family Plasma transporter B (MDR/TAP), member 1 Membrane ABCC3 2.83E-02 2.016 ATP-binding cassette, sub-family Plasma transporter C (CFTR/MRP), member 3 Membrane ABHD6 7.79E-03 −2.717 abhydrolase domain containing 6 Cytoplasm enzyme ACAT1 4.10E-02 3.009 acetyl-CoA acetyltransferase 1 Cytoplasm enzyme ACBD4 2.66E-03 1.722 acyl-CoA binding domain unknown other containing 4 ACSL5 1.86E-02 −2.876 acyl-CoA synthetase long-chain Cytoplasm enzyme family member 5 ADAM23 3.33E-02 −3.008 ADAM metallopeptidase domain Plasma peptidase 23 Membrane ADAM29 5.58E-03 3.463 ADAM metallopeptidase domain Plasma peptidase 29 Membrane ADAMTS17 2.67E-04 3.051 ADAM metallopeptidase with Extracellular other thrombospondin type 1 motif, 17 Space ADCYAP1R1 1.20E-02 1.848 adenylate cyclase activating Plasma G-protein polypeptide 1 (pituitary) receptor Membrane coupled type I receptor ADH6 (includes 4.02E-02 −1.845 alcohol dehydrogenase 6 (class Cytoplasm enzyme EG:130) V) AHSA2 1.54E-04 −1.6 AHA1, activator of heat shock unknown other 90kDa protein ATPase homolog 2 (yeast) AK5 3.32E-02 1.658 adenylate kinase 5 Cytoplasm kinase AK7
    [Show full text]
  • Composition of the Peptide Fraction in Human Blood Plasma: Database of Circulating Human Peptides
    Journal of Chromatography B, 726 (1999) 25±35 Composition of the peptide fraction in human blood plasma: database of circulating human peptides Rudolf Richtera,* , Peter Schulz-Knappe b , Michael Schrader b , Ludger StandkerÈ a , Michael JurgensÈ ba , Harald Tammen , Wolf-Georg Forssmann a aLower Saxony Institute for Peptide Research, Feodor-Lynen-Strasse 31, D-30625 Hannover, Germany bBioVisioN, Feodor-Lynen-Strasse 5, D-30625 Hannover, Germany Received 7 September 1998; received in revised form 28 December 1998; accepted 28 December 1998 Abstract A database was established from human hemo®ltrate (HF) that consisted of a mass database and a sequence database, with the aim of analyzing the composition of the peptide fraction in human blood. To establish a mass database, all 480 fractions of a peptide bank generated from HF were analyzed by MALDI-TOF mass spectrometry. Using this method, over 20 000 molecular masses representing native, circulating peptides were detected. Estimation of repeatedly detected masses suggests that approximately 5000 different peptides were recorded. More than 95% of the detected masses are smaller than 15 000, indicating that HF predominantly contains peptides. The sequence database contains over 340 entries from 75 different protein and peptide precursors. 55% of the entries are fragments from plasma proteins (®brinogen A 13%, albumin 10%, b2-microglobulin 8.5%, cystatin C 7%, and ®brinogen B 6%). Seven percent of the entries represent peptide hormones, growth factors and cytokines. Thirty-three percent belong to protein families such as complement factors, enzymes, enzyme inhibitors and transport proteins. Five percent represent novel peptides of which some show homology to known peptide and protein families.
    [Show full text]
  • Diagnostic Urinary Proteome Profile for Immunoglobulin a Nephropathy
    Archive of SID KIDNEY DISEASES Diagnostic Urinary Proteome Profile for Immunoglobulin A Nephropathy Shiva Samavat,1,2 Shiva Kalantari,3 Mohsen Nafar,3 Dorothea Rutishauser,4,5 Mostafa Rezaei-Tavirani,6,7 Mahmud Parvin,8 Roman A Zubarev4,5 1Department of Nephrology, Introduction. Immunoglobulin A (IgA) nephropathy, the most Shahid Labbafinejad Medical common type of glomerulonephritis, is only diagnosed by invasive Center, Shahid Beheshti kidney biopsy. Urine proteome panel might help in noninvasive University of Medical Sciences, Tehran, Iran diagnosis and also better understanding of pathogenesis of IgA 2Urology and Nephrology nephropathy. Research Center, Shahid Materials and Methods. Second mid-stream urine samples of 13 Beheshti University of Medical patients with biopsy-proven IgA nephropathy and 8 healthy controls Sciences, Tehran, Iran were investigated by means of nanoscale liquid chromatography 3Chronic Kidney Diseases Research Center, Shahid tandem mass spectrometry. Multivariate analysis of quantified Beheshti University of Medical label-free proteins was performed by the principal component Sciences, Tehran, Iran analysis and partial least squares models. 4Department of Medical Results. A total number of 493 unique proteins were quantified Biochemistry and Biophysics, by nanoscale liquid chromatography tandem mass spectrometry, Karolinska Institute, Stockholm, of which 46 proteins were considered as putative biomarkers of Sweden 5SciLifeLab, Stockholm, Sweden IgA nephropathy, after multivariate analysis and additional filter 6Department of Basic Sciences, criterion and comparing the patients and the controls. Some of the Faculty of Paramedical Sciences, significant differentially expressed proteins were CD44, glycoprotein Shahid Beheshti University of 2, vasorin, epidermal growth factor, CLM9, protocadherin, Medical Sciences, Tehran, Iran utreoglobin, dipeptidyl peptidase IV, NHL repeat-containing protein 7Proteomics Research Center, Faculty of Paramedical Sciences, 3, and SLAM family member 5.
    [Show full text]
  • Identification of Genomic Targets of Krüppel-Like Factor 9 in Mouse Hippocampal
    Identification of Genomic Targets of Krüppel-like Factor 9 in Mouse Hippocampal Neurons: Evidence for a role in modulating peripheral circadian clocks by Joseph R. Knoedler A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy (Neuroscience) in the University of Michigan 2016 Doctoral Committee: Professor Robert J. Denver, Chair Professor Daniel Goldman Professor Diane Robins Professor Audrey Seasholtz Associate Professor Bing Ye ©Joseph R. Knoedler All Rights Reserved 2016 To my parents, who never once questioned my decision to become the other kind of doctor, And to Lucy, who has pushed me to be a better person from day one. ii Acknowledgements I have a huge number of people to thank for having made it to this point, so in no particular order: -I would like to thank my adviser, Dr. Robert J. Denver, for his guidance, encouragement, and patience over the last seven years; his mentorship has been indispensable for my growth as a scientist -I would also like to thank my committee members, Drs. Audrey Seasholtz, Dan Goldman, Diane Robins and Bing Ye, for their constructive feedback and their willingness to meet in a frequently cold, windowless room across campus from where they work -I am hugely indebted to Pia Bagamasbad and Yasuhiro Kyono for teaching me almost everything I know about molecular biology and bioinformatics, and to Arasakumar Subramani for his tireless work during the home stretch to my dissertation -I am grateful for the Neuroscience Program leadership and staff, in particular
    [Show full text]
  • Increased Alveolar Soluble Annexin V Promotes Lung Inflammation and Fibrosis
    Increased alveolar soluble Annexin V promotes lung inflammation and fibrosis Supplemental data - Materials and methods Human BALF. Bronchoalveolar Lavage (BALF) was performed at the National Institute of Respiratory Diseases (INER), Mexico City, Mexico,as approved by the Ethics Committee at INER. is performed through flexible fiberoptic bronchoscopy under local anesthesia. Briefly, 200 ml of normal saline is instilled in 50-ml aliquots, with an average recovery of 60%–70%. The cells are pelleted from the BALF, and the cell-free supernatant is frozen at -80oC. The control group for Figure 1 included 3 healthy volunteers, and 2 individuals without HRCT-scan evidence of lung disease who required a bronchoscopy for the diagnosis of unexplained cough. In all cases bronchoscopy did not display any abnormality. Removal of Annexin V from BALF by immunoprecipitation. BALF samples with known elevated Annexin V levels were pre-cleared by mixing with 1g rabbit IgG and 20l of protein A/G agarose at 4o for 30 mins, followed by centrifuging at 1000g for 30s at 4oC. The supernatants were removed to clean tubes and incubated with mixing for 1h with 4g rabbit anti-Annexin V antibody or 4g rabbit IgG, then. 20l of protein A/G agarose was added and the tubes were incubated with mixing for 1h at 40C. The tubes were centrifuged for 30s at 1000g, and the supernatant carefully removed. PBS washes of the immunoprecipitate were pooled with the supernatant. Successful reduction of Annexin V levels was confirmed by ELISA. Intra-tracheal instillation of Annexin V solution. 8 week C57 female black mice were anaesthetized with isofluorane and suspended at a 45-60°C angle on an intubation platform.
    [Show full text]
  • The Detection of Circulating Breast Cancer Cells in Blood a M Gilbey, D Burnett, R E Coleman, I Holen
    903 REVIEW J Clin Pathol: first published as 10.1136/jcp.2003.013755 on 27 August 2004. Downloaded from The detection of circulating breast cancer cells in blood A M Gilbey, D Burnett, R E Coleman, I Holen ............................................................................................................................... J Clin Pathol 2004;57:903–911. doi: 10.1136/jcp.2003.013755 At present, sampling of the lymph nodes or bone marrow demand for supportive, palliative, and medical services. for the detection of regions of metastatic disease in patients with breast cancer can only be undertaken at the time of BREAST CANCER METASTASES initial diagnosis and surgery. However, the sampling of The metastatic process these tissues and the methods used are inaccurate, time Metastasis is a cascade of linked sequential steps involving multiple host–tumour interactions.4 consuming, and cannot be used for easy routine screening Cancer cells possessing multiple genetic abnorm- to determine disease recurrence and response to treatment. alities grow unregulated and eventually lose the Because of the problems encountered with current methods ability to adhere to one another.4 Thus, tumour cells detach from the primary tumour, migrate and tissues sampled at the time of breast cancer diagnosis, through the basement membrane and extracel- this review discusses the urgent requirement for and lular matrix, intravasate, and travel in the potential development of a quick, simple, and accurate lymphatic and/or blood systems to a new site, before attachment, extravasation, and the devel- diagnostic test utilising the haematogenous system, a opment of a new focus and neovascularistion.5 source of circulating tumour cells in patients with breast When cancer cells enter the lymphatic system, cancer, and highly sensitive molecular biological they travel to the lymph nodes (LN)4—in the case of breast cancer, to the sentinel nodes in the techniques, such as reverse transcription polymerase chain axilla and intercostal spaces—before entering the reaction.
    [Show full text]
  • Neutrophil Recruitment and Phagocytosis of Boar Spermatozoa
    REPRODUCTIONRESEARCH Effect of low-dose mifepristone administration on day 2 after ovulation on transcript profiles in implantation-stage endometrium of rhesus monkeys D Ghosh, A M Sharkey1, D S Charnock-Jones2, S K Smith3 and Jayasree Sengupta Department of Physiology, All India Institute of Medical Sciences, New Delhi 110029, India, Departments of 1Pathology and 2Obstetrics and Gynaecology, University of Cambridge, Cambridge CB2 2SW, UK and 3Faculty of Medicine, Imperial College London, London SW7 2AZ, UK Correspondence should be addressed to D Ghosh; Email: [email protected] Abstract Progesterone is essential for endometrial receptivity in primates. In studies previously performed using global gene profiling based on microarray technology, attempts have been made to identify changes in gene expression between early luteal-phase and mid-luteal-phase endometria. However, the issue of the putative impact of preimplantation embryo-derived signal in the process of endometrial receptivity was missing in the previous studies. In the present study, an attempt has been made to delineate the transcripts profile in implantation-stage endometrium under combinatorial regulation of progesterone and embryo-derived signal in the rhesus monkey. To this effect, we have compared transcript profiles for 409 known genes between control receptive stage (nZ13), and mifepristone-induced desynchronized and non-receptive stage (nZ12) monkey endometrial samples collected on days 4 (nZ12) and 6 (nZ13) after ovulation from mated, potential conception cycles,
    [Show full text]
  • Article Urinary Angiotensinogen and Risk of Severe
    Article Urinary Angiotensinogen and Risk of Severe AKI Joseph L. Alge,* Nithin Karakala,*† Benjamin A. Neely,* Michael G. Janech,*† James A. Tumlin,‡ Lakhmir S. Chawla,§ | Andrew D. Shaw, ¶ and John M. Arthur,*† for the SAKInet Investigators Summary Background Biomarkers of AKI that can predict which patients will develop severe renal disease at the time of diagnosis will facilitate timely intervention in populations at risk of adverse outcomes. *Medical University Design, setting, participants, & measurements Liquid chromatography/tandem mass spectrometry was used to of South Carolina, identify 30 potential prognostic urinary biomarkers of severe AKI in a group of patients that developed AKI after Charleston, South cardiac surgery. Angiotensinogen had the best discriminative characteristics. Urinary angiotensinogen was Carolina; †Ralph H. subsequently measured by ELISA and its prognostic predictive power was verified in 97 patients who underwent Johnson Veterans cardiac surgery between August 1, 2008 and October 6, 2011. Affairs Medical Center, Charleston, South Carolina; Results The urine angiotensinogen/creatinine ratio (uAnCR) predicted worsening of AKI, Acute Kidney Injury ‡University of Network (AKIN) stage 3, need for renal replacement therapy, discharge .7 days from sample collection, and Tennessee College of Medicine, composite outcomes of AKIN stage 2 or 3, AKIN stage 3 or death, and renal replacement therapy or death. The Chattanooga, prognostic predictive power of uAnCR was improved when only patients classified as AKIN stage 1 at the time of Tennessee; §George urine sample collection (n=79) were used in the analysis, among whom it predicted development of stage 3 AKI or Washington death with an area under the curve of 0.81.
    [Show full text]