DOCSLIB.ORG

Id4 Functions Downstream of Bmp Signaling to Restrict TCF Function in Endocardial Cells During Atrioventricular Valve Development

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Id4 Functions Downstream of Bmp Signaling to Restrict TCF Function in Endocardial Cells During Atrioventricular Valve Development Id4 functions downstream of Bmp signaling to restrict TCF function in endocardial cells during atrioventricular valve development Dissertation zur Erlangung des Doktorgrades der Naturwissenschaften vorgelegt beim Fachbereich 15 der Johann Wolfgang Goethe-Universität in Frankfurt am Main Von Suchit Ahuja aus Neu Delhi, Indien Frankfurt 2016 (D30) Vom Fachbereich der Goethe Universität als Dissertation angenommen. Dekan: Gutachter: Datum der Disputation: SUPERVISED BY Dr. Sven Reischauer, Ph.D. Department of Developmental Genetics Max Planck Institute for Heart and Lung Research Bad Nauheim, Germany REVIEWER Prof. Dr. Didier Stainier, Ph.D. Department of Developmental Genetics Max Planck Institute for Heart and Lung Research Bad Nauheim, Germany and Prof. Dr. Anna Starzinski-Powitz, Ph.D. Department of Molecular Cell Biology and Human Genetics Institute of Cell Biology and Neuroscience Johann Wolfgang Goethe University Frankfurt am Main, Germany ERKLÄRUNG Ich erkläre hiermit, dass ich mich bisher keiner Doktorprüfung im Mathematisch- Naturwissenschaftlichen Bereich unterzogen habe. Frankfurt am Main, den ............................................ (Unterschrift) Versicherung Ich erkläre hiermit, dass ich die vorgelegte Dissertation über Id4 functions downstream of Bmp signaling to restrict TCF function in endocardial cells during atrioventricular valve development selbständig angefertigt und mich anderer Hilfsmittel als der in ihr angegebenen nicht bedient habe, insbesondere, dass alle Entlehnungen aus anderen Schriften mit Angabe der betreffenden Schrift gekennzeichnet sind. Ich versichere, die Grundsätze der guten wissenschaftlichen Praxis beachtet, und nicht die Hilfe einer kommerziellen Promotionsvermittlung in Anspruch genommen zu haben. Vorliegende Ergebnisse der Arbeit sind in folgendem Publikationsorgan veröffentlicht: Ahuja S, et al. Dev. Biol. 2016; 412(1): 71-82. Id4 functions downstream of Bmp signaling to restrict TCF function in endocardial cells during atrioventricular valve development. Frankfurt am Main, den ............................................ (Unterschrift) You have to dream before your dreams can come true. Man needs his difficulties because they are necessary to enjoy success. Great dreams of great dreamers are always transcended. A.P.J. Abdul Kalam Aerospace Scientist 1931-2015 Contents Table of Contents Abbreviations ............................................................................................. 11 1. Introduction ........................................................................................... 14 1.1 Congenital Heart Disease .......................................................................... 14 1.2 Heart Valve Development ......................................................................... 16 1.2.1 Origin Of Valvular Cells: Contribution From Different Heart Layers .......................... 17 1.2.2 Endocardial Cushion Formation: Signaling Molecules Involved .................................. 17 1.3 Zebrafish Heart Development .................................................................... 22 1.4 Zebrafish Atrioventricular Canal/Valve Development................................. 25 1.5 Inhibitor of DNA Binding 4 (Id4) .............................................................. 27 1.6 AIM of the Project .................................................................................... 29 2. Materials ................................................................................................... 30 2.1 Antibiotics ................................................................................................ 30 2.2 Antibodies ................................................................................................ 30 2.3 Bacterial Culture medium .......................................................................... 31 2.4 Bacterial Strains ........................................................................................ 31 2.5 Buffers/Solutions ...................................................................................... 31 2.6 Centrifuges ............................................................................................... 33 2.7 Chemicals ................................................................................................. 34 2.8 Enzymes ................................................................................................... 36 2.9 Kits .......................................................................................................... 37 2.10 Microscopes ............................................................................................. 37 2.11 Miscellaneous Equipment .......................................................................... 38 2.12 Oligonucleotides ....................................................................................... 40 2.13 PCR enzymes and mastermix .................................................................... 41 2.14 Plasmids ................................................................................................... 42 2.15 Softwares .................................................................................................. 43 Contents 2.16 Tubes and containers ................................................................................. 43 2.17 Zebrafish Food .......................................................................................... 44 2.18 Zebrafish Lines ......................................................................................... 44 3. Methods ..................................................................................................... 45 3.1 Zebrafish Husbandry ................................................................................. 45 3.1.1 Zebrafish Maintenance .................................................................................................. 45 3.1.2 Zebrafish Breeding ........................................................................................................ 45 3.2 Zebrafish Microinjection ........................................................................... 45 3.2.1 Preparation of Injection plates ....................................................................................... 45 3.2.2 Preparation of Injection Needles ................................................................................... 46 3.2.3 Microinjection ............................................................................................................... 46 3.3 RNA Isolation ........................................................................................... 46 3.4 Reverse Transcription or cDNA Synthesis ................................................. 46 3.5 PCR amplification from cDNA .................................................................. 47 3.6 Agarose Gel Electrophoresis ...................................................................... 47 3.7 PCR Product Elution From Agarose Gel .................................................... 48 3.8 E.Coli Competent Cells Preparation ........................................................... 48 3.9 Transformation of Competent E.Coli ......................................................... 48 3.10 DNA Restriction Digestion ........................................................................ 48 3.11 Cloning .................................................................................................... 49 3.11.1 TA Cloning .................................................................................................................... 49 3.11.2 Cold Fusion ................................................................................................................... 49 3.11.3 Cloning id4 Overexpression Constructs ........................................................................ 50 3.12 Plasmid DNA Isolation ............................................................................. 50 3.13 In situ Hybridisation .................................................................................. 50 3.13.1 Probe Synthesis ............................................................................................................. 50 3.13.2 Embryo/Adult Heart preparation for in situ hybridization ............................................ 51 3.13.3 In situ Hybidization Day 1: Probe Hybridization .......................................................... 51 3.13.4 In situ Hybidization Day 2: Unbound Probe Removal .................................................. 52 3.13.5 In situ Hybidization Day 3: Unbound Antibody Removal and Staining ....................... 53 3.13.6 In situ Hybidization: Microscopy and Imaging ............................................................. 53 Contents 3.14 TALEN (Transcription Activator-Like Effector Nucleases) Induced Mutagenesis ....................................................................................................... 53 3.14.1 TALEN Designing......................................................................................................... 53 3.14.2 TALEN Assembly Day 1 .............................................................................................. 54 3.14.3 TALEN Assembly Day 2 .............................................................................................. 54 3.14.4 TALEN Assembly Day 3 .............................................................................................. 55 3.14.5 TALEN Assembly Day 4 .............................................................................................
Load more

Recommended publications
  • Uterine Double-Conditional Inactivation of Smad2 and Smad3 in Mice Causes Endometrial Dysregulation, Infertility, and Uterine Cancer
    Uterine double-conditional inactivation of Smad2 and Smad3 in mice causes endometrial dysregulation, infertility, and uterine cancer Maya Krisemana,b, Diana Monsivaisa,c, Julio Agnoa, Ramya P. Masanda, Chad J. Creightond,e, and Martin M. Matzuka,c,f,g,h,1 aDepartment of Pathology and Immunology, Baylor College of Medicine, Houston, TX 77030; bReproductive Endocrinology and Infertility, Baylor College of Medicine/Texas Children’s Hospital Women’s Pavilion, Houston, TX 77030; cCenter for Drug Discovery, Baylor College of Medicine, Houston, TX 77030; dDepartment of Medicine, Baylor College of Medicine, Houston, TX 77030; eDan L. Duncan Comprehensive Cancer Center, Baylor College of Medicine, Houston, TX 77030; fDepartment of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX 77030; gDepartment of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX 77030; and hDepartment of Pharmacology and Chemical Biology, Baylor College of Medicine, Houston, TX 77030 Contributed by Martin M. Matzuk, December 6, 2018 (sent for review April 30, 2018; reviewed by Milan K. Bagchi and Thomas E. Spencer) SMAD2 and SMAD3 are downstream proteins in the transforming in endometrial function. Notably, members of the transforming growth factor-β (TGF β) signaling pathway that translocate signals growth factor β (TGF β) family are involved in many cellular from the cell membrane to the nucleus, bind DNA, and control the processes and serve as principal regulators of numerous biological expression of target genes. While SMAD2/3 have important roles functions, including female reproduction. Previous studies have in the ovary, we do not fully understand the roles of SMAD2/3 in shown the TGF β family to have key roles in ovarian folliculo- the uterus and their implications in the reproductive system.
    [Show full text]
  • Table S1 the Four Gene Sets Derived from Gene Expression Profiles of Escs and Differentiated Cells
    Table S1 The four gene sets derived from gene expression profiles of ESCs and differentiated cells Uniform High Uniform Low ES Up ES Down EntrezID GeneSymbol EntrezID GeneSymbol EntrezID GeneSymbol EntrezID GeneSymbol 269261 Rpl12 11354 Abpa 68239 Krt42 15132 Hbb-bh1 67891 Rpl4 11537 Cfd 26380 Esrrb 15126 Hba-x 55949 Eef1b2 11698 Ambn 73703 Dppa2 15111 Hand2 18148 Npm1 11730 Ang3 67374 Jam2 65255 Asb4 67427 Rps20 11731 Ang2 22702 Zfp42 17292 Mesp1 15481 Hspa8 11807 Apoa2 58865 Tdh 19737 Rgs5 100041686 LOC100041686 11814 Apoc3 26388 Ifi202b 225518 Prdm6 11983 Atpif1 11945 Atp4b 11614 Nr0b1 20378 Frzb 19241 Tmsb4x 12007 Azgp1 76815 Calcoco2 12767 Cxcr4 20116 Rps8 12044 Bcl2a1a 219132 D14Ertd668e 103889 Hoxb2 20103 Rps5 12047 Bcl2a1d 381411 Gm1967 17701 Msx1 14694 Gnb2l1 12049 Bcl2l10 20899 Stra8 23796 Aplnr 19941 Rpl26 12096 Bglap1 78625 1700061G19Rik 12627 Cfc1 12070 Ngfrap1 12097 Bglap2 21816 Tgm1 12622 Cer1 19989 Rpl7 12267 C3ar1 67405 Nts 21385 Tbx2 19896 Rpl10a 12279 C9 435337 EG435337 56720 Tdo2 20044 Rps14 12391 Cav3 545913 Zscan4d 16869 Lhx1 19175 Psmb6 12409 Cbr2 244448 Triml1 22253 Unc5c 22627 Ywhae 12477 Ctla4 69134 2200001I15Rik 14174 Fgf3 19951 Rpl32 12523 Cd84 66065 Hsd17b14 16542 Kdr 66152 1110020P15Rik 12524 Cd86 81879 Tcfcp2l1 15122 Hba-a1 66489 Rpl35 12640 Cga 17907 Mylpf 15414 Hoxb6 15519 Hsp90aa1 12642 Ch25h 26424 Nr5a2 210530 Leprel1 66483 Rpl36al 12655 Chi3l3 83560 Tex14 12338 Capn6 27370 Rps26 12796 Camp 17450 Morc1 20671 Sox17 66576 Uqcrh 12869 Cox8b 79455 Pdcl2 20613 Snai1 22154 Tubb5 12959 Cryba4 231821 Centa1 17897
    [Show full text]
  • The Role of Neural Crest in Vertebrate Evolution
    The Role of Neural Crest in Vertebrate Evolution: Tissue-specific Genetic Labeling in Zebrafish (Danio rerio) Reveals Neural Crest Contribution to Gills, but Not to Scales Dissertation der Mathematisch-Naturwissenschaftlichen Fakultät der Eberhard Karls Universität Tübingen zur Erlangung des Grades eines Doktors der Naturwissenschaften (Dr. rer. nat.) vorgelegt von Alessandro Mongera aus Trento (Italien) Tübingen 2013 Tag der mündlichen Qualifikation: 09.12.2013 Dekan: Prof. Dr. Wolfgang Rosenstiel 1. Berichterstatter: Prof. Dr. Christiane Nüsslein-Volhard 2. Berichterstatter: Prof. Dr. Rolf Reuter 3. Berichterstatter: Dr. Didier Stainier to my friends Contents Abstract 7 Zusammenfassung 11 1 Introduction 15 1.1 Neural crest as a source of vertebrate innovations 15 1.2 Tools for cell lineage tracing . 18 1.2.1 Non-genetic labeling . 18 1.2.2 Cre/loxP-mediated genetic labeling . 21 1.3 The gills . 23 1.4 The post-cranial integumentary skeleton . 25 2 Results 29 Publication 1 . 29 Publication 2 . 43 3 Discussion 49 3.1 NC-driven expansion of the respiratory surface 49 3.2 Mesoderm, and not NC, contributes to fish scales 52 3.3 Tg(sox10:ERT2-Cre) line as a tool to under- stand vertebrate evolution . 55 Bibliography 56 5 Own contribution to the manuscripts 69 Curriculum vitae 71 Full publication list 73 Acknowledgments 75 Appendix 77 Publication 3 . 79 Publication 4 . 95 6 Abstract During vertebrate evolution, the development of an active, predatory lifestyle and an increased body size coincided with the appearance of many morphological innovations. Among these vertebrate-specific phenotypic features, a major role in promoting the massive radiation of this animal group has been played by a new, true head.
    [Show full text]
  • Co-Occupancy by Multiple Cardiac Transcription Factors Identifies
    Co-occupancy by multiple cardiac transcription factors identifies transcriptional enhancers active in heart Aibin Hea,b,1, Sek Won Konga,b,c,1, Qing Maa,b, and William T. Pua,b,2 aDepartment of Cardiology and cChildren’s Hospital Informatics Program, Children’s Hospital Boston, Boston, MA 02115; and bHarvard Stem Cell Institute, Harvard University, Cambridge, MA 02138 Edited by Eric N. Olson, University of Texas Southwestern, Dallas, TX, and approved February 23, 2011 (received for review November 12, 2010) Identification of genomic regions that control tissue-specific gene study of a handful of model genes (e.g., refs. 7–10), it has not been expression is currently problematic. ChIP and high-throughput se- evaluated using unbiased, genome-wide approaches. quencing (ChIP-seq) of enhancer-associated proteins such as p300 In this study, we used a modified ChIP-seq approach to define identifies some but not all enhancers active in a tissue. Here we genome wide the binding sites of these cardiac TFs (1). We show that co-occupancy of a chromatin region by multiple tran- provide unbiased support for collaborative TF interactions in scription factors (TFs) identifies a distinct set of enhancers. GATA- driving cardiac gene expression and use this principle to show that chromatin co-occupancy by multiple TFs identifies enhancers binding protein 4 (GATA4), NK2 transcription factor-related, lo- with cardiac activity in vivo. The majority of these multiple TF- cus 5 (NKX2-5), T-box 5 (TBX5), serum response factor (SRF), and “ binding loci (MTL) enhancers were distinct from p300-bound myocyte-enhancer factor 2A (MEF2A), here referred to as cardiac enhancers in location and functional properties.
    [Show full text]
  • Jdp2 Downregulates Trp53 Transcription to Promote Leukaemogenesis in the Context of Trp53 Heterozygosity
    Oncogene (2013) 32, 397 --402 & 2013 Macmillan Publishers Limited All rights reserved 0950-9232/13 www.nature.com/onc SHORT COMMUNICATION Jdp2 downregulates Trp53 transcription to promote leukaemogenesis in the context of Trp53 heterozygosity L van der Weyden1, AG Rust1,4, RE McIntyre1,4, CD Robles-Espinoza1, M del Castillo Velasco-Herrera1, R Strogantsev2, AC Ferguson-Smith2, S McCarthy1, TM Keane1, MJ Arends3 and DJ Adams1 We performed a genetic screen in mice to identify candidate genes that are associated with leukaemogenesis in the context of Trp53 heterozygosity. To do this we generated Trp53 heterozygous mice carrying the T2/Onc transposon and SB11 transposase alleles to allow transposon-mediated insertional mutagenesis to occur. From the resulting leukaemias/lymphomas that developed in these mice, we identified nine loci that are potentially associated with tumour formation in the context of Trp53 heterozygosity, including AB041803 and the Jun dimerization protein 2 (Jdp2). We show that Jdp2 transcriptionally regulates the Trp53 promoter, via an atypical AP-1 site, and that Jdp2 expression negatively regulates Trp53 expression levels. This study is the first to identify a genetic mechanism for tumour formation in the context of Trp53 heterozygosity. Oncogene (2013) 32, 397--402; doi:10.1038/onc.2012.56; published online 27 February 2012 Keywords: p53; Jdp2; transposon; heterozygosity; lymphoma; mice INTRODUCTION targeted by transposon insertion events leading to upregulated Genetic alterations of TP53 are frequent events in tumourigenesis Jdp2 expression and a decrease in Trp53 expression levels. Further and promote genomic instability, impair apoptosis, and contribute we illustrate that Jdp2 regulates the Trp53 promoter via an atypical to aberrant self-renewal.1--4 The spectrum of mutations that occur AP-1 binding site.
    [Show full text]
  • HAND2‑Mediated Proteolysis Negatively Regulates the Function of Estrogen Receptor Α
    5538 MOLECULAR MEDICINE REPORTS 12: 5538-5544, 2015 HAND2‑mediated proteolysis negatively regulates the function of estrogen receptor α TOMOHIKO FUKUDA*, AKIRA SHIRANE*, OSAMU WADA-HIRAIKE, KATSUTOSHI ODA, MICHIHIRO TANIKAWA, AYAKO SAKUABASHI, MANA HIRANO, HOUJU FU, YOSHIHIRO MORITA, YUICHIRO MIYAMOTO, KANAKO INABA, KEI KAWANA, YUTAKA OSUGA and TOMOYUKI FUJII Department of Obstetrics and Gynecology, Graduate School of Medicine, The University of Tokyo, Tokyo 113-8655, Japan Received October 2, 2014; Accepted June 11, 2015 DOI: 10.3892/mmr.2015.4070 Abstract. A previous study demonstrated that the transcriptional activation function of ERα was possibly attrib- progesterone-inducible HAND2 gene product is a basic uted to the proteasomic degradation of ERα by HAND2. These helix-loop-helix transcription factor and prevents mitogenic results indicate a novel anti-tumorigenic function of HAND2 effects of estrogen receptor α (ERα) by inhibiting fibroblast in regulating ERα-dependent gene expression. Considering growth factor signalling in mouse uteri. However, whether that HAND2 is commonly hypermethylated and silenced in HAND2 directly affects the transcriptional activation func- endometrial cancer, it is hypothesized that HAND2 may serve tion of ERα remains to be elucidated. In the present study, as a possible tumor suppressor, particularly in uterine tissue. the physical interaction between HAND2 and ERα was investigating by performing an immunoprecipitation assay Introduction and an in vitro pull-down assay. The results demonstrated that HAND2 and ERα interacted in a ligand-independent manner. Endometrial cancer is one of the most common types of The in vitro pull-down assays revealed a direct interaction gynecologic malignancy, increasing each year (1). Based on between HAND2 and the amino-terminus of ERα, termed a pathological view, endometrial cancer can be divided into the activation function-1 domain.
    [Show full text]
  • A Dissertation Entitled the Androgen Receptor
    A Dissertation entitled The Androgen Receptor as a Transcriptional Co-activator: Implications in the Growth and Progression of Prostate Cancer By Mesfin Gonit Submitted to the Graduate Faculty as partial fulfillment of the requirements for the PhD Degree in Biomedical science Dr. Manohar Ratnam, Committee Chair Dr. Lirim Shemshedini, Committee Member Dr. Robert Trumbly, Committee Member Dr. Edwin Sanchez, Committee Member Dr. Beata Lecka -Czernik, Committee Member Dr. Patricia R. Komuniecki, Dean College of Graduate Studies The University of Toledo August 2011 Copyright 2011, Mesfin Gonit This document is copyrighted material. Under copyright law, no parts of this document may be reproduced without the expressed permission of the author. An Abstract of The Androgen Receptor as a Transcriptional Co-activator: Implications in the Growth and Progression of Prostate Cancer By Mesfin Gonit As partial fulfillment of the requirements for the PhD Degree in Biomedical science The University of Toledo August 2011 Prostate cancer depends on the androgen receptor (AR) for growth and survival even in the absence of androgen. In the classical models of gene activation by AR, ligand activated AR signals through binding to the androgen response elements (AREs) in the target gene promoter/enhancer. In the present study the role of AREs in the androgen- independent transcriptional signaling was investigated using LP50 cells, derived from parental LNCaP cells through extended passage in vitro. LP50 cells reflected the signature gene overexpression profile of advanced clinical prostate tumors. The growth of LP50 cells was profoundly dependent on nuclear localized AR but was independent of androgen. Nevertheless, in these cells AR was unable to bind to AREs in the absence of androgen.
    [Show full text]
  • Transcriptional Adaptation in Caenorhabditis Elegans
    RESEARCH ARTICLE Transcriptional adaptation in Caenorhabditis elegans Vahan Serobyan1*, Zacharias Kontarakis1†, Mohamed A El-Brolosy1, Jordan M Welker1, Oleg Tolstenkov2,3‡, Amr M Saadeldein1, Nicholas Retzer1, Alexander Gottschalk2,3,4, Ann M Wehman5, Didier YR Stainier1* 1Department of Developmental Genetics, Max Planck Institute for Heart and Lung Research, Bad Nauheim, Germany; 2Institute for Biophysical Chemistry, Goethe University, Frankfurt Am Main, Germany; 3Cluster of Excellence Frankfurt - Macromolecular Complexes (CEF-MC), Goethe University, Frankfurt Am Main, Germany; 4Buchmann Institute for Molecular Life Sciences (BMLS), Goethe University, Frankfurt Am Main, Germany; 5Rudolf Virchow Center, University of Wu¨ rzburg, Wu¨ rzburg, Germany Abstract Transcriptional adaptation is a recently described phenomenon by which a mutation in *For correspondence: one gene leads to the transcriptional modulation of related genes, termed adapting genes. At the [email protected] molecular level, it has been proposed that the mutant mRNA, rather than the loss of protein (VS); function, activates this response. While several examples of transcriptional adaptation have been [email protected] (DYRS) reported in zebrafish embryos and in mouse cell lines, it is not known whether this phenomenon is observed across metazoans. Here we report transcriptional adaptation in C. elegans, and find that † Present address: Genome this process requires factors involved in mutant mRNA decay, as in zebrafish and mouse. We Engineering and Measurement further uncover a requirement for Argonaute proteins and Dicer, factors involved in small RNA Lab, ETH Zurich, Functional maturation and transport into the nucleus. Altogether, these results provide evidence for Genomics Center Zurich of ETH transcriptional adaptation in C.
    [Show full text]
  • Regulation of Runx2 Accumulation and Its Consequences
    Regulation of Runx2 Accumulation and Its Consequences Junko Shimazu Submitted in partial fulfillment of the requirement for the degree of Doctor of Philosophy in the Graduate School of Arts and Sciences COLUMBIA UNIVERSITY 2016 ©2016 Junko Shimazu All Rights Reserved ABSTRACT Regulation of Runx2 Accumulation and Its Consequences Junko Shimazu Osteoblasts are bone-forming cells and therefore they are responsible of the synthesis of type I collagen, the main component of bone matrix. However, there is an apparent disconnect between the regulation of osteoblast differentiation and bone formation since the synthesis of Type I collagen precedes the expression of Runx2, the earliest determinant of osteoblast differentiation. Recently, genetic experiments in the mouse have revealed the existence of an unexpected cross-regulation between bone and other organs. In particular this body of work has highlighted the importance of osteoblasts as endocrine cells to regulate whole-body glucose homeostasis by secretion of a hormone, osteocalcin. However, the fundamental question of why bone regulates glucose homeostasis remained to be answered. Therefore, in my thesis, considering that bone is a metabolically demanding organ that constantly renews itself, I hypothesized that characterizing the connection between the need of glucose as a main nutrient in osteoblasts and bone development will provide a key to deeper understanding of why bone regulates glucose homeostasis. My work shows here that glucose uptake through GLUT1 in osteoblasts is needed for osteoblast differentiation by suppressing the AMPK-dependent activation by phosphorylation at S148 of Smurf1 that targets Runx2 for degradation. I also uncovered the mechanism of action of Smurf1 in this setting.
    [Show full text]
  • Genetic Factors in Nonsyndromic Orofacial Clefts
    Published online: 2021-02-12 THIEME Review Article 101 Genetic Factors in Nonsyndromic Orofacial Clefts Mahamad Irfanulla Khan1 Prashanth C.S.2 Narasimha Murthy Srinath3 1 Department of Orthodontics & Dentofacial Orthopedics, The Oxford Address for correspondence Mahamad Irfanulla Khan, BDS, MDS, Dental College, Bangalore, Karnataka, India Department of Orthodontics & Dentofacial Orthopedics, The Oxford 2 Department of Orthodontics & Dentofacial Orthopedics, DAPM R.V. Dental College, Bangalore, Karnataka 560068, India Dental College, Bangalore, Karnataka, India (e-mail: [email protected]). 3 Department of Oral & Maxillofacial Surgery, Krishnadevaraya College of Dental Sciences, Bangalore, Karnataka, India Global Med Genet 2020;7:101–108. Abstract Orofacial clefts (OFCs) are the most common congenital birth defects in humans and immediately recognized at birth. The etiology remains complex and poorly understood and seems to result from multiple genetic and environmental factors along with gene– environment interactions. It can be classified into syndromic (30%) and nonsyndromic (70%) clefts. Nonsyndromic OFCs include clefts without any additional physical or Keywords cognitive deficits. Recently, various genetic approaches, such as genome-wide associ- ► orofacial clefts ation studies (GWAS), candidate gene association studies, and linkage analysis, have ► nonsyndromic identified multiple genes involved in the etiology of OFCs. ► genetics This article provides an insight into the multiple genes involved in the etiology of OFCs. ► gene mutation Identification of specific genetic causes of clefts helps in a better understanding of the ► genome-wide molecular pathogenesis of OFC. In the near future, it helps to provide a more accurate association study diagnosis, genetic counseling, personalized medicine for better clinical care, and ► linkage analysis prevention of OFCs.
    [Show full text]
  • Effects of Estrogen Receptor and Wnt Signaling Activation On
    International Journal of Molecular Sciences Article Effects of Estrogen Receptor and Wnt Signaling Activation on Mechanically Induced Bone Formation in a Mouse Model of Postmenopausal Bone Loss 1, , 1, 1 1 Astrid Liedert * y, Claudia Nemitz y, Melanie Haffner-Luntzer , Fabian Schick , Franz Jakob 2 and Anita Ignatius 1 1 Institute of Orhopedic Research and Biomechanics, Trauma Research Center Ulm, University Medical Center Ulm, 89081 Ulm, Germany; [email protected] (C.N.); melanie.haff[email protected] (M.H.-L.); [email protected] (F.S.); [email protected] (A.I.) 2 Orthopaedic Center for Musculoskeletal Research, University of Würzburg, 97074 Würzburg, Germany; [email protected] * Correspondence: [email protected]; Tel.: +49-731-500-55333; Fax: +49-731-500-55302 These authors contributed equally to the study. y Received: 14 September 2020; Accepted: 31 October 2020; Published: 5 November 2020 Abstract: In the adult skeleton, bone remodeling is required to replace damaged bone and functionally adapt bone mass and structure according to the mechanical requirements. It is regulated by multiple endocrine and paracrine factors, including hormones and growth factors, which interact in a coordinated manner. Because the response of bone to mechanical signals is dependent on functional estrogen receptor (ER) and Wnt/β-catenin signaling and is impaired in postmenopausal osteoporosis by estrogen deficiency, it is of paramount importance to elucidate the underlying mechanisms as a basis for the development of new strategies in the treatment of osteoporosis. The present study aimed to investigate the effectiveness of the activation of the ligand-dependent ER and the Wnt/β-catenin signal transduction pathways on mechanically induced bone formation using ovariectomized mice as a model of postmenopausal bone loss.
    [Show full text]
  • Supplementary Information
    Osa et al Supplementary Information Clinical implications of monitoring nivolumab immunokinetics in previously treated non– small cell lung cancer patients Akio Osa, Takeshi Uenami, Shohei Koyama, Kosuke Fujimoto, Daisuke Okuzaki, Takayuki Takimoto, Haruhiko Hirata, Yukihiro Yano, Soichiro Yokota, Yuhei Kinehara, Yujiro Naito, Tomoyuki Otsuka, Masaki Kanazu, Muneyoshi Kuroyama, Masanari Hamaguchi, Taro Koba, Yu Futami, Mikako Ishijima, Yasuhiko Suga, Yuki Akazawa, Hirotomo Machiyama, Kota Iwahori, Hyota Takamatsu, Izumi Nagatomo, Yoshito Takeda, Hiroshi Kida, Esra A. Akbay, Peter S. Hammerman, Kwok-kin Wong, Glenn Dranoff, Masahide Mori, Takashi Kijima, Atsushi Kumanogoh Supplemental Figures 1 – 8 1 Osa et al Supplemental Figure 1. The frequency of nivolumab-bound T cells was maintained in patients who continued treatment. Nivolumab binding in CD8 and CD4 T cells was analyzed at two follow-up points, as indicated, in fresh peripheral blood from three representative cases from protocol 1 that continued treatment. 2 Osa et al Supplemental Figure 2. Long-term follow-up of nivolumab binding to T cells from fresh whole blood. Nivolumab binding was followed up in fresh peripheral blood from an additional case, Pt.7. 3 Osa et al Supplemental Figure 3. Long-term duration of nivolumab binding is due to sustained circulation of residual nivolumab in plasma. (A) PBMCs acquired from Pt.8 and 9 at pretreatment (pre PBMCs) and after a single dose (post 1 PBMCs) were cultured in regular medium without nivolumab (top and middle). Pre PBMCs were also incubated with 10 µg/ml nivolumab in vitro before the cultures were started (bottom). Nivolumab binding status was monitored at the indicated time points.
    [Show full text]