DOCSLIB.ORG

View of Untreated Versus DT Treated Glomeruli at 5 Days

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
View of Untreated Versus DT Treated Glomeruli at 5 Days BASIC RESEARCH www.jasn.org Proximal Tubules Have the Capacity to Regulate Uptake of Albumin † Mark C. Wagner,* Silvia B. Campos-Bilderback,* Mahboob Chowdhury, Brittany Flores,* ‡ Xianyin Lai, Jered Myslinski,* Sweekar Pandit,* Ruben M. Sandoval,* Sarah E. Wean,* † | Yuan Wei,§ Lisa M. Satlin,§ Roger C. Wiggins, Frank A. Witzmann, and | Bruce A. Molitoris* *Indiana University School of Medicine, The Roudebush Veterans Affair Medical Center, Indiana Center for Biological Microscopy, Indianapolis, Indiana; §Department of Pediatrics, The Icahn School of Medicine at Mount Sinai, New York; †Department of Internal Medicine, University of Michigan, Ann Arbor, Michigan; ‡Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, Indianapolis, Indiana; and |Department of Cellular and Integrative Physiology, Indiana University School of Medicine, Indianapolis, Indiana ABSTRACT Evidence from multiple studies supports the concept that both glomerular filtration and proximal tubule (PT) reclamation affect urinary albumin excretion rate. To better understand these roles of glomerular filtration and PT uptake, we investigated these processes in two distinct animal models. In a rat model of acute exogenous albumin overload, we quantified glomerular sieving coefficients (GSC) and PT uptake of Texas Red-labeled rat serum albumin using two-photon intravital microscopy. No change in GSC was observed, but a significant decrease in PT albumin uptake was quantified. In a second model, loss of endogenous albumin was induced in rats by podocyte-specific transgenic expression of diphtheria toxin receptor. In these albumin-deficient rats, exposure to diphtheria toxin induced an increase in albumin GSC and albumin filtration, resulting in increased exposure of the PTs to endogenous albumin. In this case, PT albumin reabsorption was markedly increased. Analysis of known albumin receptors and assessment of cortical protein expression in the albumin overload model, conducted to identify potential proteins and pathways affected by acute protein overload, revealed changes in the expression levels of calreticulin, disabled homolog 2, NRF2, angiopoietin-2, and proteins involved in ATP synthesis. Taken together, these results suggest that a regulated PT cell albumin uptake system can respond rapidly to different physiologic conditions to minimize alterations in serum albumin level. J Am Soc Nephrol 27: 482–494, 2016. doi: 10.1681/ASN.2014111107 While the clinical relevance of proteinuria, and disease conditions. This understanding is important especially albuminuria, has been well documented, because before appropriate therapies can be devel- the quantitative and mechanistic significance of oped to modulate albuminuria, the structural, func- different components to albumin excretion remains tional and mechanistic characterization need to be an area of considerable excitement and debate.1 Recent better understood and interrelated. data from several laboratories utilizing different ap- proaches have delineated a role of proximal tubules (PTs) in regulation of albumin reabsorption and rec- Received November 18, 2014. Accepted May 4, 2015. 2 lamation. Furthermore, albumin transcytosis has Published online ahead of print. Publication date available at 3 been visualized in PT cells and FcRn has been shown www.jasn.org. to mediate the transcytosis of albumin across PTCs,4 Correspondence: Dr. Bruce A. Molitoris, Nephrology Division, 2 as it is known to do for many other cell types. There- Department of Medicine, Indiana University School of Medicine, fore, the present studies were conducted to begin dif- 950 West Walnut Street, Building R2, Room 266, Indianapolis, IN ferentiating and quantifying glomerular and PT 46202. Email: [email protected] contributions to albuminuria under physiologic and Copyright © 2016 by the American Society of Nephrology 482 ISSN : 1046-6673/2702-482 J Am Soc Nephrol 27: 482–494, 2016 www.jasn.org BASIC RESEARCH PTs regulate glomerular filtrate uptake, sorting, degradation, albumin from baseline. An average of 75% of the amount of transcytosis, and secretion, thus serving as a key contributor in albumin injected was recovered in the urine on days 2–5. This determining urine composition. Further, PT cells have extremely increase in albuminuria is consistent with previous results, which active endocytosis mechanisms and are thus responsible also showed no change in hematocrit or total plasma protein.9 for retrieving most filtered proteins under normal conditions.5 Glomerular filtration rate (GFR, ml/min/100 g) measured using Given the recent data supporting increased filtration of proteins 24-hour creatinine clearances, showed no significant measurable (i.e., albumin), understanding their endocytic mechanisms be- change between day 0 and 3 or 4 day 1=0.7160.14, imaging comes even more important.2 A clear role of megalin/cubilin in day=0.6860.12. receptor-mediated clathrin-dependent endocytosis function ex- ists and in addition there is an active non-selective fluid-phase Acute Albumin Overload does not Correlate with endocytosis pathway.6 However, the only known transcytosis re- Glomerular Sieving Alterations ceptor for albumin is FcRn.7 While many questions remain con- This acute albumin overload model allowed us to determine cerning these processes it is clear that multiple diseases result whether elevated albumin excretion resulted from increased from protein trafficking defects in PT.2,8 Therefore, to begin ad- passageviatheglomerularfiltrationbarrier.Intravitaltwo-photon dressing how the PTs handle filtered proteins two different rat microscopy was used to quantify GSCs on days 3 or 4, enabling us models of proteinuria have been investigated. to evaluate GSC for rats with variable but elevated albuminuria First, the acute albumin overload model, which has been used levels. Each rat had an average of five surface glomeruli that were for over 20 years and induces a significant rapid and reversible measured at two time points following Texas Red rat serum increase in albuminuria, was evaluated.9,10 The second model albumin(TR-RSA)infusion.Imagecollectiondidnotstartuntilat was a transgenic rat line that specifically expresses the diphtheria least 10 minutes post TR-RSA infusion to allow equilibration to toxin (DT) receptor in podocytes under the control of the podocyte- occur. Figure 1D shows the GSC values on the y axis (overall specific podocin promoter. Under physiologic conditions urinary mean of 0.008060.0013) and the corresponding 24-hour urine albumin was unaltered, but the presence of DTresults in selective albumin values plotted on the x axis. Note, these values were also podocyte injury/death resulting in increased albumin leak across no different from GSCs for control MWF female rats (mean GSC the glomerular filtration barrier.11 of 0.007560.0029). If increased passage of albumin across the An implicit assumption has always been that in both models glomerular filtration barrier was the cause of albuminuria then albuminuria results from enhanced passage of albumin across the one would expect that rats with the highest albumin excretion glomerular filtration barrier. However, recent data from multiple levels would have the highest GSC values. However, there was no laboratories supports a role for both the glomerulus and PT cell in correlation between the measured GSC for albumin and the determining the level of albuminuria in disease processes.2 Pre- 24-hour urinary albumin excretion. This result raised the possi- viously it had not been possible to evaluate the role of the PTC in bility that a nonglomerular event was responsible for increased these models. However, intravital two-photon microscopy of the albumin excretion. The other well-studied mechanism that has kidney now permits a direct evaluation of changes occurring in been shown to contribute to elevated urinary albumin is an al- glomerular filtration and PTC uptake of fluorescent molecules teration in PTC uptake of filtered proteins.2 simultaneously in the same nephron.12,13 An increase in the glo- merular sieving coefficient (GSC) would be consistent with a PT Albumin Reabsorption Decreased During Albumin glomerular effect while changes in the level of PTC uptake would Loading support a role for the PTC in albuminuria. The ability to simul- Intravital microscopy was used to quantify tracer amounts of TR- taneously monitor both mechanisms is essential for understand- RSAuptake by PTCs. Less TR-RSAappeared in the early S1 segment ing the mechanism of albuminuria. of the PTs (visible opening to glomerulus) in protein overload rats compared with control rats. An example of this is shown in Figure 2A where the control PT S1 had increased TR-RSA fluorescence RESULTS than a PT S1 in the protein overloaded rat, images captured 20–30 min after TR-RSA infusion. To quantify TR-RSA uptake we used Albuminuria in the MWF Rats Following Albumin our established method that determines total PTC uptake without Overload distinguishing between different PT segments, Figure 2B.12 Abox Initial studies were undertaken to characterize the albumin plot of these data gave a mean value of 335761339 total integrated overload model in the female Munich Wistar Fromter (MWF) fluorescence (TIF)/mm2 for control and 12556976 TIF/mm2 for rats. Their superficial glomeruli have allowed our laboratory, and protein overload with a P value of 0.014. To further characterize others, to directly observe and quantify glomerular and tubular uptake changes we focused on the robust endocytic activity of the events
Load more

Recommended publications
  • Proteomic Profile of Human Spermatozoa in Healthy And
    Cao et al. Reproductive Biology and Endocrinology (2018) 16:16 https://doi.org/10.1186/s12958-018-0334-1 REVIEW Open Access Proteomic profile of human spermatozoa in healthy and asthenozoospermic individuals Xiaodan Cao, Yun Cui, Xiaoxia Zhang, Jiangtao Lou, Jun Zhou, Huafeng Bei and Renxiong Wei* Abstract Asthenozoospermia is considered as a common cause of male infertility and characterized by reduced sperm motility. However, the molecular mechanism that impairs sperm motility remains unknown in most cases. In the present review, we briefly reviewed the proteome of spermatozoa and seminal plasma in asthenozoospermia and considered post-translational modifications in spermatozoa of asthenozoospermia. The reduction of sperm motility in asthenozoospermic patients had been attributed to factors, for instance, energy metabolism dysfunction or structural defects in the sperm-tail protein components and the differential proteins potentially involved in sperm motility such as COX6B, ODF, TUBB2B were described. Comparative proteomic analysis open a window to discover the potential pathogenic mechanisms of asthenozoospermia and the biomarkers with clinical significance. Keywords: Proteome, Spermatozoa, Sperm motility, Asthenozoospermia, Infertility Background fertilization failure [4] and it has become clear that iden- Infertility is defined as the lack of ability to achieve a tifying the precise proteins and the pathways involved in clinical pregnancy after one year or more of unprotected sperm motility is needed [5]. and well-timed intercourse with the same partner [1]. It is estimated that around 15% of couples of reproductive age present with infertility, and about half of the infertil- Application of proteomic techniques in male ity is associated with male partner [2, 3].
    [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]
  • 4-6 Weeks Old Female C57BL/6 Mice Obtained from Jackson Labs Were Used for Cell Isolation
    Methods Mice: 4-6 weeks old female C57BL/6 mice obtained from Jackson labs were used for cell isolation. Female Foxp3-IRES-GFP reporter mice (1), backcrossed to B6/C57 background for 10 generations, were used for the isolation of naïve CD4 and naïve CD8 cells for the RNAseq experiments. The mice were housed in pathogen-free animal facility in the La Jolla Institute for Allergy and Immunology and were used according to protocols approved by the Institutional Animal Care and use Committee. Preparation of cells: Subsets of thymocytes were isolated by cell sorting as previously described (2), after cell surface staining using CD4 (GK1.5), CD8 (53-6.7), CD3ε (145- 2C11), CD24 (M1/69) (all from Biolegend). DP cells: CD4+CD8 int/hi; CD4 SP cells: CD4CD3 hi, CD24 int/lo; CD8 SP cells: CD8 int/hi CD4 CD3 hi, CD24 int/lo (Fig S2). Peripheral subsets were isolated after pooling spleen and lymph nodes. T cells were enriched by negative isolation using Dynabeads (Dynabeads untouched mouse T cells, 11413D, Invitrogen). After surface staining for CD4 (GK1.5), CD8 (53-6.7), CD62L (MEL-14), CD25 (PC61) and CD44 (IM7), naïve CD4+CD62L hiCD25-CD44lo and naïve CD8+CD62L hiCD25-CD44lo were obtained by sorting (BD FACS Aria). Additionally, for the RNAseq experiments, CD4 and CD8 naïve cells were isolated by sorting T cells from the Foxp3- IRES-GFP mice: CD4+CD62LhiCD25–CD44lo GFP(FOXP3)– and CD8+CD62LhiCD25– CD44lo GFP(FOXP3)– (antibodies were from Biolegend). In some cases, naïve CD4 cells were cultured in vitro under Th1 or Th2 polarizing conditions (3, 4).
    [Show full text]
  • Supplemental Information
    Supplemental information Dissection of the genomic structure of the miR-183/96/182 gene. Previously, we showed that the miR-183/96/182 cluster is an intergenic miRNA cluster, located in a ~60-kb interval between the genes encoding nuclear respiratory factor-1 (Nrf1) and ubiquitin-conjugating enzyme E2H (Ube2h) on mouse chr6qA3.3 (1). To start to uncover the genomic structure of the miR- 183/96/182 gene, we first studied genomic features around miR-183/96/182 in the UCSC genome browser (http://genome.UCSC.edu/), and identified two CpG islands 3.4-6.5 kb 5’ of pre-miR-183, the most 5’ miRNA of the cluster (Fig. 1A; Fig. S1 and Seq. S1). A cDNA clone, AK044220, located at 3.2-4.6 kb 5’ to pre-miR-183, encompasses the second CpG island (Fig. 1A; Fig. S1). We hypothesized that this cDNA clone was derived from 5’ exon(s) of the primary transcript of the miR-183/96/182 gene, as CpG islands are often associated with promoters (2). Supporting this hypothesis, multiple expressed sequences detected by gene-trap clones, including clone D016D06 (3, 4), were co-localized with the cDNA clone AK044220 (Fig. 1A; Fig. S1). Clone D016D06, deposited by the German GeneTrap Consortium (GGTC) (http://tikus.gsf.de) (3, 4), was derived from insertion of a retroviral construct, rFlpROSAβgeo in 129S2 ES cells (Fig. 1A and C). The rFlpROSAβgeo construct carries a promoterless reporter gene, the β−geo cassette - an in-frame fusion of the β-galactosidase and neomycin resistance (Neor) gene (5), with a splicing acceptor (SA) immediately upstream, and a polyA signal downstream of the β−geo cassette (Fig.
    [Show full text]
  • Identification of Differentially Expressed Genes in Human Bladder Cancer Through Genome-Wide Gene Expression Profiling
    521-531 24/7/06 18:28 Page 521 ONCOLOGY REPORTS 16: 521-531, 2006 521 Identification of differentially expressed genes in human bladder cancer through genome-wide gene expression profiling KAZUMORI KAWAKAMI1,3, HIDEKI ENOKIDA1, TOKUSHI TACHIWADA1, TAKENARI GOTANDA1, KENGO TSUNEYOSHI1, HIROYUKI KUBO1, KENRYU NISHIYAMA1, MASAKI TAKIGUCHI2, MASAYUKI NAKAGAWA1 and NAOHIKO SEKI3 1Department of Urology, Graduate School of Medical and Dental Sciences, Kagoshima University, 8-35-1 Sakuragaoka, Kagoshima 890-8520; Departments of 2Biochemistry and Genetics, and 3Functional Genomics, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba 260-8670, Japan Received February 15, 2006; Accepted April 27, 2006 Abstract. Large-scale gene expression profiling is an effective CKS2 gene not only as a potential biomarker for diagnosing, strategy for understanding the progression of bladder cancer but also for staging human BC. This is the first report (BC). The aim of this study was to identify genes that are demonstrating that CKS2 expression is strongly correlated expressed differently in the course of BC progression and to with the progression of human BC. establish new biomarkers for BC. Specimens from 21 patients with pathologically confirmed superficial (n=10) or Introduction invasive (n=11) BC and 4 normal bladder samples were studied; samples from 14 of the 21 BC samples were subjected Bladder cancer (BC) is among the 5 most common to microarray analysis. The validity of the microarray results malignancies worldwide, and the 2nd most common tumor of was verified by real-time RT-PCR. Of the 136 up-regulated the genitourinary tract and the 2nd most common cause of genes we detected, 21 were present in all 14 BCs examined death in patients with cancer of the urinary tract (1-7).
    [Show full text]
  • In This Table Protein Name, Uniprot Code, Gene Name P-Value
    Supplementary Table S1: In this table protein name, uniprot code, gene name p-value and Fold change (FC) for each comparison are shown, for 299 of the 301 significantly regulated proteins found in both comparisons (p-value<0.01, fold change (FC) >+/-0.37) ALS versus control and FTLD-U versus control. Two uncharacterized proteins have been excluded from this list Protein name Uniprot Gene name p value FC FTLD-U p value FC ALS FTLD-U ALS Cytochrome b-c1 complex P14927 UQCRB 1.534E-03 -1.591E+00 6.005E-04 -1.639E+00 subunit 7 NADH dehydrogenase O95182 NDUFA7 4.127E-04 -9.471E-01 3.467E-05 -1.643E+00 [ubiquinone] 1 alpha subcomplex subunit 7 NADH dehydrogenase O43678 NDUFA2 3.230E-04 -9.145E-01 2.113E-04 -1.450E+00 [ubiquinone] 1 alpha subcomplex subunit 2 NADH dehydrogenase O43920 NDUFS5 1.769E-04 -8.829E-01 3.235E-05 -1.007E+00 [ubiquinone] iron-sulfur protein 5 ARF GTPase-activating A0A0C4DGN6 GIT1 1.306E-03 -8.810E-01 1.115E-03 -7.228E-01 protein GIT1 Methylglutaconyl-CoA Q13825 AUH 6.097E-04 -7.666E-01 5.619E-06 -1.178E+00 hydratase, mitochondrial ADP/ATP translocase 1 P12235 SLC25A4 6.068E-03 -6.095E-01 3.595E-04 -1.011E+00 MIC J3QTA6 CHCHD6 1.090E-04 -5.913E-01 2.124E-03 -5.948E-01 MIC J3QTA6 CHCHD6 1.090E-04 -5.913E-01 2.124E-03 -5.948E-01 Protein kinase C and casein Q9BY11 PACSIN1 3.837E-03 -5.863E-01 3.680E-06 -1.824E+00 kinase substrate in neurons protein 1 Tubulin polymerization- O94811 TPPP 6.466E-03 -5.755E-01 6.943E-06 -1.169E+00 promoting protein MIC C9JRZ6 CHCHD3 2.912E-02 -6.187E-01 2.195E-03 -9.781E-01 Mitochondrial 2-
    [Show full text]
  • Definition of the Landscape of Promoter DNA Hypomethylation in Liver Cancer
    Published OnlineFirst July 11, 2011; DOI: 10.1158/0008-5472.CAN-10-3823 Cancer Therapeutics, Targets, and Chemical Biology Research Definition of the Landscape of Promoter DNA Hypomethylation in Liver Cancer Barbara Stefanska1, Jian Huang4, Bishnu Bhattacharyya1, Matthew Suderman1,2, Michael Hallett3, Ze-Guang Han4, and Moshe Szyf1,2 Abstract We use hepatic cellular carcinoma (HCC), one of the most common human cancers, as a model to delineate the landscape of promoter hypomethylation in cancer. Using a combination of methylated DNA immunopre- cipitation and hybridization with comprehensive promoter arrays, we have identified approximately 3,700 promoters that are hypomethylated in tumor samples. The hypomethylated promoters appeared in clusters across the genome suggesting that a high-level organization underlies the epigenomic changes in cancer. In normal liver, most hypomethylated promoters showed an intermediate level of methylation and expression, however, high-CpG dense promoters showed the most profound increase in gene expression. The demethylated genes are mainly involved in cell growth, cell adhesion and communication, signal transduction, mobility, and invasion; functions that are essential for cancer progression and metastasis. The DNA methylation inhibitor, 5- aza-20-deoxycytidine, activated several of the genes that are demethylated and induced in tumors, supporting a causal role for demethylation in activation of these genes. Previous studies suggested that MBD2 was involved in demethylation of specific human breast and prostate cancer genes. Whereas MBD2 depletion in normal liver cells had little or no effect, we found that its depletion in human HCC and adenocarcinoma cells resulted in suppression of cell growth, anchorage-independent growth and invasiveness as well as an increase in promoter methylation and silencing of several of the genes that are hypomethylated in tumors.
    [Show full text]
  • Blood-Derived Mitochondrial DNA Copy Number Is Associated with Gene Expression Across Multiple
    bioRxiv preprint doi: https://doi.org/10.1101/2020.07.17.209023; this version posted July 18, 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. 1 Blood-derived mitochondrial DNA copy number is associated with gene expression across multiple 2 tissues and is predictive for incident neurodegenerative disease 3 4 Stephanie Y. Yang1, Christina A. Castellani1, Ryan J. Longchamps1, Vamsee K. Pillalamarri1, Brian 5 O’Rourke2, Eliseo Guallar3, Dan E. Arking1,2 6 7 8 1McKusick-Nathans Department of Genetic Medicine, Johns Hopkins University School of Medicine, 9 Baltimore, MD 10 2Division of Cardiology, Department of Medicine, Johns Hopkins University School of Medicine, 11 Baltimore, MD 12 3Departments of Epidemiology and Medicine, and Welch Center for Prevention, Epidemiology, and 13 Clinical Research, Johns Hopkins University Bloomberg School of Public Health, Baltimore, MD 14 15 Email addresses: 16 [email protected] 17 [email protected] 18 [email protected] 19 [email protected] 20 [email protected] 21 [email protected] 22 [email protected] 23 24 25 26 27 28 29 30 31 32 33 34 35 *Correspondence and address for reprints to: 36 Dan E. Arking, Ph.D. 37 Johns Hopkins University School of Medicine 38 733 N Broadway Ave 39 Miller Research Building Room 459 40 Baltimore, MD 21205 41 (410) 502-4867 (ph) 42 [email protected] 1 bioRxiv preprint doi: https://doi.org/10.1101/2020.07.17.209023; this version posted July 18, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder.
    [Show full text]
  • Bypass Budget Proposal for Fiscal Year 2020 �
    Open Science, � Big Data, and You � Working Together to Treat and Prevent Alzheimer’s Disease and Related Dementias NIH BYPASS BUDGET PROPOSAL FOR FISCAL YEAR 2020 � National Institutes of Health CONTENTS OPEN SCIENCE, BIG DATA, AND YOU: WORKING TOGETHER TO TREAT AND PREVENT ALZHEIMER’S DISEASE AND RELATED DEMENTIAS ....................................................................... 1 INTRODUCTION .............................................................................................................................. 3 A Sense of Urgency ..................................................................................................................... 4 The Nation’s Commitment Becomes Law ................................................................................... 5 Increasing and Targeting the Investment in Research ............................................................... 6 Budgeting in FY 2020 to Fight Dementia .................................................................................... 7 Together, We Make the Difference .......................................................................................... 11 FISCAL YEAR 2020 PROFESSIONAL JUDGMENT BUDGET: ALZHEIMER’S DISEASE AND RELATED DEMENTIAS ................................................................................................................................... 12 THE ROAD TO TREATMENT AND PREVENTION OF ALZHEIMER’S DISEASE AND RELATED DEMENTIAS ..................................................................................................................................
    [Show full text]
  • A High-Throughput Approach to Uncover Novel Roles of APOBEC2, a Functional Orphan of the AID/APOBEC Family
    Rockefeller University Digital Commons @ RU Student Theses and Dissertations 2018 A High-Throughput Approach to Uncover Novel Roles of APOBEC2, a Functional Orphan of the AID/APOBEC Family Linda Molla Follow this and additional works at: https://digitalcommons.rockefeller.edu/ student_theses_and_dissertations Part of the Life Sciences Commons A HIGH-THROUGHPUT APPROACH TO UNCOVER NOVEL ROLES OF APOBEC2, A FUNCTIONAL ORPHAN OF THE AID/APOBEC FAMILY A Thesis Presented to the Faculty of The Rockefeller University in Partial Fulfillment of the Requirements for the degree of Doctor of Philosophy by Linda Molla June 2018 © Copyright by Linda Molla 2018 A HIGH-THROUGHPUT APPROACH TO UNCOVER NOVEL ROLES OF APOBEC2, A FUNCTIONAL ORPHAN OF THE AID/APOBEC FAMILY Linda Molla, Ph.D. The Rockefeller University 2018 APOBEC2 is a member of the AID/APOBEC cytidine deaminase family of proteins. Unlike most of AID/APOBEC, however, APOBEC2’s function remains elusive. Previous research has implicated APOBEC2 in diverse organisms and cellular processes such as muscle biology (in Mus musculus), regeneration (in Danio rerio), and development (in Xenopus laevis). APOBEC2 has also been implicated in cancer. However the enzymatic activity, substrate or physiological target(s) of APOBEC2 are unknown. For this thesis, I have combined Next Generation Sequencing (NGS) techniques with state-of-the-art molecular biology to determine the physiological targets of APOBEC2. Using a cell culture muscle differentiation system, and RNA sequencing (RNA-Seq) by polyA capture, I demonstrated that unlike the AID/APOBEC family member APOBEC1, APOBEC2 is not an RNA editor. Using the same system combined with enhanced Reduced Representation Bisulfite Sequencing (eRRBS) analyses I showed that, unlike the AID/APOBEC family member AID, APOBEC2 does not act as a 5-methyl-C deaminase.
    [Show full text]
  • Proteomic Shifts in Embryonic Stem Cells with Gene Dose Modifications Suggest the Presence of Balancer Proteins in Protein Regulatory Networks
    Proteomic shifts in embryonic stem cells with gene dose modifications suggest the presence of balancer proteins in protein regulatory networks. Lei Mao, Claus Zabel, Marion Herrmann, Tobias Nolden, Florian Mertes, Laetitia Magnol, Caroline Chabert, Daniela Hartl, Yann Herault, Jean Maurice Delabar, et al. To cite this version: Lei Mao, Claus Zabel, Marion Herrmann, Tobias Nolden, Florian Mertes, et al.. Proteomic shifts in embryonic stem cells with gene dose modifications suggest the presence of balancer proteins in protein regulatory networks.. PLoS ONE, Public Library of Science, 2007, 2 (11), pp.e1218. 10.1371/jour- nal.pone.0001218. hal-00408296 HAL Id: hal-00408296 https://hal.archives-ouvertes.fr/hal-00408296 Submitted on 31 May 2020 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Proteomic Shifts in Embryonic Stem Cells with Gene Dose Modifications Suggest the Presence of Balancer Proteins in Protein Regulatory Networks Lei Mao1,2*, Claus Zabel1, Marion Herrmann1, Tobias Nolden2, Florian Mertes2, Laetitia Magnol3, Caroline Chabert4, Daniela
    [Show full text]
  • Dominant Mutations in Mtdna Maintenance Gene SSBP1 Cause Optic Atrophy and Foveopathy
    Dominant mutations in mtDNA maintenance gene SSBP1 cause optic atrophy and foveopathy Camille Piro-Mégy, … , Maria Sola, Cécile Delettre J Clin Invest. 2020;130(1):143-156. https://doi.org/10.1172/JCI128513. Research Article Genetics Ophthalmology Graphical abstract Find the latest version: https://jci.me/128513/pdf The Journal of Clinical Investigation RESEARCH ARTICLE Dominant mutations in mtDNA maintenance gene SSBP1 cause optic atrophy and foveopathy Camille Piro-Mégy,1 Emmanuelle Sarzi,1 Aleix Tarrés-Solé,2 Marie Péquignot,1 Fenna Hensen,3 Mélanie Quilès,1 Gaël Manes,1 Arka Chakraborty,2 Audrey Sénéchal,1 Béatrice Bocquet,1,4 Chantal Cazevieille,1 Agathe Roubertie,1,4 Agnès Müller,1,5 Majida Charif,6 David Goudenège,6 Guy Lenaers,6 Helmut Wilhelm,7 Ulrich Kellner,8 Nicole Weisschuh,9 Bernd Wissinger,9 Xavier Zanlonghi,10 Christian Hamel,1,4 Johannes N. Spelbrink,3 Maria Sola,2 and Cécile Delettre1 1Institute of Neurosciences of Montpellier, INSERM, University of Montpellier, Montpellier, France. 2Structural MitoLab, Department of Structural Biology, “Maria de Maeztu” Unit of Excellence, Molecular Biology Institute Barcelona (IBMB-CSIC), Barcelona, Spain. 3Radboud Center for Mitochondrial Medicine, Department of Paediatrics, Radboudumc, Nijmegen, Netherlands. 4CHU Montpellier, Centre of Reference for Genetic Sensory Diseases, Gui de Chauliac Hospital, Montpellier, France. 5Faculté de Pharmacie, Université de Montpellier, Montpellier, France. 6UMR CNRS 6015-INSERM U1083, MitoVasc Institute, Angers University, Angers, France. 7University Eye Hospital, Centre for Ophthalmology, University of Tübingen, Tübingen, Germany. 8Rare Retinal Disease Center, AugenZentrum Siegburg, MVZ Augenärztliches Diagnostik- und Therapiecentrum Siegburg GmbH, Siegburg, Germany. 9Institute for Ophthalmic Research, Centre for Ophthalmology, University of Tübingen, Tübingen, Germany.
    [Show full text]