DOCSLIB.ORG

Original Article WNT3A Gene Expression Is Associated with Isolated Hirschsprung Disease Polymorphism and Disease Status

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Original Article WNT3A Gene Expression Is Associated with Isolated Hirschsprung Disease Polymorphism and Disease Status Int J Clin Exp Pathol 2014;7(4):1359-1368 www.ijcep.com /ISSN:1936-2625/IJCEP1401084 Original Article WNT3A gene expression is associated with isolated Hirschsprung disease polymorphism and disease status Dong Chen1, Jie Mi1, Xiaomei Liu2, Juan Zhang3, Weilin Wang1, Hong Gao2 1Department of Pediatric Surgery, Shengjing Hospital of China Medical University, Shenyang 110004, China; 2Key Laboratory of Pediatric Congenital Malformations, Ministry of Public Health, Shengjing Hospital of China Medical University, Shenyang 110004, China; 3Jinzhou Women and Children’s Hospital, Jinzhou, Liaoning, 121000, China Received January 22, 2014; Accepted March 11, 2014; Epub March 15, 2014; Published April 1, 2014 Abstract: WNT3A has been regarded as an activator of the canonical Wnt signaling pathway. It has been found Wnt signaling pathway is closely related with embrionic development and Hirschsprung disease (HSCR). A com- mon haplotype consisting of minor SNPs alleles located in the WNT3A gene has been described as a risk factor for various genetic disorders. However, whether WNT3A contributes to the onset of HSCR has not been identified. The present study aims to detect the interactions of genetic variations in the WNT3A gene and examine the biological expression levels with Hirschsprung disease (HSCR) in the Chinese people. We analyzed WNT3A gene (rs61743220, rs192966556 and rs145882986) variants in the whole blood samples from HSCR patients and normal children (control groups). WNT3A expression was also examined by quantitative real-time PCR (qRT-PCR), western blotting and immunostaining. Consequently, when rs192966556 and rs145882986 alleles of the WNT3A gene lack the SNPs, they are especially associated with a greater risk of HSCR (OR [95% confidence interval] = 1.791, p = 0.001; OR [95% confidence interval] = 1.556, p = 0.003, respectively). The mRNA and protein expressions of WNT3A were higher in the aganglionic colon segment tissues than in the normal ganglionic segments tissues. Immunostaining indicates that the staining of WNT3A was much stronger (brown) in the aganglionic colon segment tissues than that in the normal ganglionic colon segment tissues (colorless or light yellow) in the mucous layer and muscular layer. Although preliminary, these results suggest that WNT3A may play an important role in the pathogenesis of HSCR. Keywords: Hirschsprung disease, WNT3A, polymorphism, gene and protein, expression Introduction sporadic HSCR cases carry RET mutations. Besides RET and EDNRB, other genes have Hirschsprung disease (HSCR) is a congenital been identified in sporadic affected individuals, disorder of the enteric nervous system (ENS) such as endothelin 3 (EDN3) [5], glial derived and is characterized by the absence of intesti- neurotrophic factor (GDNF) [6], sex determining nal ganglion cells in myenteric and submucosal region Y-box 10 (SOX10) [7], paired-like homeo- plexuses. Its incidence is approximately 1/ box 2B (PHOX2B) [8] and ZFHX1B (SIP1) [9]. 5,000 human live births, and has a male pre- Thus, HSCR has become a model of a complex ponderance of 4:1 [1]. At present, the cause of polygenic disorder in which the interaction of HSCR still remains unclear, but there is a com- different genes need to be elucidated. mon understanding that HSCR is a complex dis- ease influenced by multiple genetic and envi- Wnt signaling is one of the most critical signal- ronmental factors. Recent studies have shown ing pathways in many aspects of development that the genetic etiology of the neurocristopa- [10, 11]. Wnt signaling is subdivided into the thy is complex and many genes may be involved canonical β-catenin dependent and the non- in the development of HSCR [2-4]. It is well canonical β-catenin independent branch. known that RET and EDNRB are primary genes Canonical Wnt signaling is characterized by the in the etiology of HSCR. The RET mutations are stabilization and subsequent nuclear transport associated with the development of HSCR in of β-catenin resulting in the activation of tran- 50% of cases in familial series, but only 3% of scriptional responses. Non-canonical Wnt sig- Polymorphism and expression of WNT3A in HSCR naling is more diverse and includes several dif- 3.5 years old including 156 males and 44 ferent signaling modes that regulate cell females (average age, 1.5 ± 0.3 years), and behavior. Genes of the Wnt family encode were recruited as the HSCR groups. An addi- structurally related, cystein-rich glycoproteins tional 200 healthy children that matched with involved in cell-cell signalling for a wide variety the HSCR group in age and gender were used of developmental processes [12-15]. Wnt can as control groups (average age, 2 ± 0.5 years). act either on the cell that secrets them (auto- The control groups had no history of constipa- crine feedback) or on neighboring cells by para- tion. Tissue samples (aganglionic and normal crine signaling. Wnt signaling pathway plays ganglionic colon segment tissues) were rather important roles in the process of the obtained from 50 HSCR patients (41 males and embryonic development. It has been shown 9 females) with pathologically confirmed HSCR that the Wnt signaling pathway plays important pre- or post-operatively at Shengjing Hospital role in intestinal cell proliferation [16]. of China Medical University. Age ranged from 0.5 to 4.5 years old with an average of 1.5 WNT3A, a canonical Wnt ligand, has been years. None of these cases received any preop- regarded as an activator of the canonical Wnt erative treatment or had a history of HSCR’s signaling pathway. WNT3A has been reported enterocolitis or active enterocolitis at the time to induce the accumulation of β-catenin and of surgery. Tissue samples were collected activation of the canonical Wnt signaling path- immediately after operation and were stored at way [17]. Mice that are homozygous for a hypo- -80°C until use. morphic wnt3a allele display vertebral defects, a short tail due to loss of caudal vertebrae, defi- Genomic DNA extraction cient cloacal development and incomplete uro- rectal septation [18]. Moreover, studies involv- Venous blood (200 µl) was obtained from the ing human pluripotent stem cells have shown study participants using EDTA as an anticoagu- that WNT3A is required for hindgut specifica- lant. Genomic DNA of peripheral blood white tion [19]. Thus, WNT3A is a pivotal component blood cells (WBCs) was extracted according to of the mesoderm gene that regulates network the QIAamp® DNA Blood Mini Kit Handbook. with ramifications for embryonic development. For the present study, the absorbance value at The objective of this study was to investigate 260/280 nm (A260/A280) ranged from 1.6 to 2.0, whether genetic variations in the WNT3A gene which met the requirements for further are associated with HSCR and examine the bio- experiments. logical expression levels of WNT3A in the Chinese people. The single nucleotide polymor- Polymerase chain reaction (PCR), DNA se- phisms (SNPs) in WNT3A gene (rs61743220, rs quence analysis and statistical analysis 192966556 and rs145882986) were selected and analyzed in a group of patients with HSCR We have chosen rs61743220, rs192966556 and matched control samples. In this study, we and rs145882986 for WNT3A gene as our poly- further detected differential expressions of morphism analysis loci based on the values of WNT3A by qRT-PCR, western blot and MAF. Specific primers to WNT3A gene were immunostaining. designed using DNASTAR primer design pro- gram and synthesized by Invitrogen Co. Materials and methods (Shanghai) (Table 1). The primers have no homology with other genes as determined by Patients and specimens BLAST analysis on homology. This study was approved by Ethics Committee Genomic DNA from peripheral blood was of China Medical University (Ethical Number: obtained with QIAamp Blood kits (Takara, 2013PS07K). Blood samples of 200 HSCR Dalian, China) using standard methods [20]. patients were collected from the Department of Basically, 150 ng of genomic DNA was ampli- Pediatric Surgery, Shengjing Hospital of China fied in a 50 μl reaction that contained 20 pmol Medical University. Patients with familial consti- each primer and 2U Taq DNA polymerase pation and a history of other congenital gastro- (TaKaRa Biotechnology Co.). Conditions for PCR intestinal tract malformations were excluded amplification in rs61743220, rs192966556 from the study. The patients ranged from 0.5 to and rs145882986 of the WNT3A gene frag- 1360 Int J Clin Exp Pathol 2014;7(4):1359-1368 Polymorphism and expression of WNT3A in HSCR Table 1. Primer sequences and conditions for polymorphism analysis value was recorded. The average CT value was the Location & Annealing SNP locus Sequence (5’-3’) Amplicon temperature extreme CT value of the sample. The expression dif- rs61743220 F: ATA CAC CAC CCA ACC TCA CG 306 bp 57°C ference of the gene was cal- R: AGA CAC CAT CCC ACC AAA CT ‘A/G’ culated by the 2-ΔΔct method rs192966556 F: AGA GCA AAG GGT CTG TAG C 237 bp 55°C [21]. R: CAC TCC TGG ATG CCA AT ‘C/T’ rs145882986 F: CAG GCA AGG GCT GGA AGT 316 bp 59°C Western blot analysis R: GCT GTG GTG TAG CTG GAT GG ‘C/T’ Approximately 50 mg speci- men was minced to small ments are shown in Table 1. A 50 µl reaction pieces using surgical blades and sonicated in system included 10 × PCR buffer solutions, 1 protein lysis buffer. Protein concentrations mmol/L of MgCl2, 0.1 mmol/L of dNTP, 10 were measured by the Bradford method, and pmol/L of each of the upstream and down- specimens were adjusted to the same protein stream primers, 100 ng of DNA template, and concentration, aliquoted and stored at -80°C. 1U of Taq DNA polymerase. PCR condition was: Equal amounts of total proteins from tissues denaturing at 95°C for 3 min, then 35 cycles of were separated on SDS-polyacrylamide gels denaturing at 95°C for 30 sec, annealing at and then electro-transfer to PVDF membranes 55~59°C for 30 sec and elongation at 72°C for (Millipore, USA).
Load more

Recommended publications
  • Detailed Review Paper on Retinoid Pathway Signalling
    1 1 Detailed Review Paper on Retinoid Pathway Signalling 2 December 2020 3 2 4 Foreword 5 1. Project 4.97 to develop a Detailed Review Paper (DRP) on the Retinoid System 6 was added to the Test Guidelines Programme work plan in 2015. The project was 7 originally proposed by Sweden and the European Commission later joined the project as 8 a co-lead. In 2019, the OECD Secretariat was added to coordinate input from expert 9 consultants. The initial objectives of the project were to: 10 draft a review of the biology of retinoid signalling pathway, 11 describe retinoid-mediated effects on various organ systems, 12 identify relevant retinoid in vitro and ex vivo assays that measure mechanistic 13 effects of chemicals for development, and 14 Identify in vivo endpoints that could be added to existing test guidelines to 15 identify chemical effects on retinoid pathway signalling. 16 2. This DRP is intended to expand the recommendations for the retinoid pathway 17 included in the OECD Detailed Review Paper on the State of the Science on Novel In 18 vitro and In vivo Screening and Testing Methods and Endpoints for Evaluating 19 Endocrine Disruptors (DRP No 178). The retinoid signalling pathway was one of seven 20 endocrine pathways considered to be susceptible to environmental endocrine disruption 21 and for which relevant endpoints could be measured in new or existing OECD Test 22 Guidelines for evaluating endocrine disruption. Due to the complexity of retinoid 23 signalling across multiple organ systems, this effort was foreseen as a multi-step process.
    [Show full text]
  • WNT11-Conditioned Medium Promotes Angiogenesis Through the Activation of Non-Canonical WNT-PKC-JNK Signaling Pathway
    G C A T T A C G G C A T genes Article WNT11-Conditioned Medium Promotes Angiogenesis through the Activation of Non-Canonical WNT-PKC-JNK Signaling Pathway § Jingcai Wang y, Min Gong z, Shi Zuo , Jie Xu, Chris Paul, Hongxia Li k, Min Liu, Yi-Gang Wang, Muhammad Ashraf ¶ and Meifeng Xu * Department of Pathology and Laboratory Medicine, University of Cincinnati Medical Center, Cincinnati, OH 45267, USA; [email protected] (J.W.); [email protected] (M.G.); [email protected] (S.Z.); [email protected] (J.X.); [email protected] (C.P.); [email protected] (H.L.); [email protected] (M.L.); [email protected] (Y.-G.W.); [email protected] (M.A.) * Correspondence: [email protected] Current address: Department of Pathology and Laboratory Medicine, Nationwide Children’s Hospital, y Columbus, OH 43205, USA. Current Address: Department of Neonatology, Children’s Hospital of Soochow University, z Suzhou 215025, Jiangsu, China. § Current Address: Department of Hepatobiliary Surgery, The Affiliated Hospital of Guizhou Medical University, Guiyang 550025, Guizhou, China. Current Address: Department of Cardiology, The First Affiliated Hospital of Soochow University, k Suzhou 215006, Jiangsu, China. ¶ Current Address: Department of Medicine, Cardiology, Medical College of Georgia, Augusta University, Augusta, GA 30912, USA. Received: 10 August 2020; Accepted: 26 October 2020; Published: 29 October 2020 Abstract: Background: We demonstrated that the transduction of Wnt11 into mesenchymal stem cells (MSCs) (MSCWnt11) promotes these cells differentiation into cardiac phenotypes. In the present study, we investigated the paracrine effects of MSCWnt11 on cardiac function and angiogenesis.
    [Show full text]
  • WNT4 and WNT3A Activate Cell Autonomous Wnt Signaling Independent of Secretion
    bioRxiv preprint doi: https://doi.org/10.1101/333906; this version posted September 14, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC 4.0 International license. Running Title: Secretion-independent Wnt signaling Research article WNT4 and WNT3A activate cell autonomous Wnt signaling independent of secretion Deviyani M. Rao1, Rebecca L. Ferguson1, Tomomi M. Yamamoto2, Benjamin G. Bitler2, Matthew J. Sikora1 Affiliation: 1Dept. of Pathology, 2Dept. of Obstetrics and Gynecology, University of Colorado Anschutz Medical Campus Corresponding author: Matthew J. Sikora, PhD.; Mail Stop 8104, Research Complex 1 South, Room 5117, 12801 E. 17th Ave.; Aurora, CO 80045. Tel: (303)724-4301; Fax: (303)724-3712; email: [email protected]. Twitter: @mjsikora Funding This work was supported by R00 CA193734 (MJS) and R00 CA194318 (BGB) from the National Institutes of Health, and by a grant from the Cancer League of Colorado, Inc (MJS). Authors' contributions DMR and MJS conceived of the project and experiments. DMR, RLF, and MJS designed and performed experiments. RLF, DMR, and TMY developed models for the project. DMR, RLF, BGB, and MJS contributed to data analysis and interpretation. DMR wrote the draft manuscript; all authors read and revised the manuscript, and have read and approved of this version of the manuscript. bioRxiv preprint doi: https://doi.org/10.1101/333906; this version posted September 14, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity.
    [Show full text]
  • Wnt/Β-Catenin Signaling Regulates Regeneration in Diverse Tissues of the Zebrafish
    Wnt/β-catenin Signaling Regulates Regeneration in Diverse Tissues of the Zebrafish Nicholas Stockton Strand A dissertation Submitted in partial fulfillment of the Requirements for the degree of Doctor of Philosophy University of Washington 2016 Reading Committee: Randall Moon, Chair Neil Nathanson Ronald Kwon Program Authorized to Offer Degree: Pharmacology ©Copyright 2016 Nicholas Stockton Strand University of Washington Abstract Wnt/β-catenin Signaling Regulates Regeneration in Diverse Tissues of the Zebrafish Nicholas Stockton Strand Chair of the Supervisory Committee: Professor Randall T Moon Department of Pharmacology The ability to regenerate tissue after injury is limited by species, tissue type, and age of the organism. Understanding the mechanisms of endogenous regeneration provides greater insight into this remarkable biological process while also offering up potential therapeutic targets for promoting regeneration in humans. The Wnt/β-catenin signaling pathway has been implicated in zebrafish regeneration, including the fin and nervous system. The body of work presented here expands upon the role of Wnt/β-catenin signaling in regeneration, characterizing roles for Wnt/β-catenin signaling in multiple tissues. We show that cholinergic signaling is required for blastema formation and Wnt/β-catenin signaling initiation in the caudal fin, and that overexpression of Wnt/β-catenin ligand is sufficient to rescue blastema formation in fins lacking cholinergic activity. Next, we characterized the glial response to Wnt/β-catenin signaling after spinal cord injury, demonstrating that Wnt/β-catenin signaling is necessary for recovery of motor function and the formation of bipolar glia after spinal cord injury. Lastly, we defined a role for Wnt/β-catenin signaling in heart regeneration, showing that cardiomyocyte proliferation is regulated by Wnt/β-catenin signaling.
    [Show full text]
  • Wnt-Independent and Wnt-Dependent Effects of APC Loss on the Chemotherapeutic Response
    International Journal of Molecular Sciences Review Wnt-Independent and Wnt-Dependent Effects of APC Loss on the Chemotherapeutic Response Casey D. Stefanski 1,2 and Jenifer R. Prosperi 1,2,3,* 1 Department of Biological Sciences, University of Notre Dame, Notre Dame, IN 46617, USA; [email protected] 2 Mike and Josie Harper Cancer Research Institute, South Bend, IN 46617, USA 3 Department of Biochemistry and Molecular Biology, Indiana University School of Medicine-South Bend, South Bend, IN 46617, USA * Correspondence: [email protected]; Tel.: +1-574-631-4002 Received: 30 September 2020; Accepted: 20 October 2020; Published: 22 October 2020 Abstract: Resistance to chemotherapy occurs through mechanisms within the epithelial tumor cells or through interactions with components of the tumor microenvironment (TME). Chemoresistance and the development of recurrent tumors are two of the leading factors of cancer-related deaths. The Adenomatous Polyposis Coli (APC) tumor suppressor is lost in many different cancers, including colorectal, breast, and prostate cancer, and its loss correlates with a decreased overall survival in cancer patients. While APC is commonly known for its role as a negative regulator of the WNT pathway, APC has numerous binding partners and functional roles. Through APC’s interactions with DNA repair proteins, DNA replication proteins, tubulin, and other components, recent evidence has shown that APC regulates the chemotherapy response in cancer cells. In this review article, we provide an overview of some of the cellular processes in which APC participates and how they impact chemoresistance through both epithelial- and TME-derived mechanisms. Keywords: adenomatous polyposis coli; chemoresistance; WNT signaling 1.
    [Show full text]
  • Genome‐Wide Analysis of Canonical Wnt Target Gene Regulation in Xenopus Tropicalis Challenges Β‐Catenin Paradigm
    Review genesis DOI 10.1002/dvg.22991 Title Genome-wide analysis of canonical Wnt target gene regulation in Xenopus tropicalis challenges βcatenin paradigm Authors Yukio Nakamura and Stefan Hoppler Affiliations Institute of Medical Sciences, Foresterhill Health Campus, University of Aberdeen, Aberdeen AB25 2ZD, UK Telephone +44 1224 437383 Email [email protected] Running head Context-specific Wnt target gene regulation Keywords Wnt signaling; βcatenin; Xenopus; gastrula; ChIP-seq; RNA-seq This article has been accepted for publication and undergone full peer review but has not been through the copyediting, typesetting, pagination and proofreading process which may lead to differences between this version and the Version of Record. Please cite this article as an ‘Accepted Article’, doi: 10.1002/dvg.22991 © 2017 Wiley Periodicals, Inc. Received: Oct 27, 2016; Accepted: Oct 30, 2016 This article is protected by copyright. All rights reserved. genesis Page 2 of 21 Abstract Wnt/β-catenin signaling is an important cell-to-cell signaling mechanism that controls gene expression during embryonic development and is critically implicated in human diseases. Developmental, cellular, and transcriptional responses to Wnt signaling are remarkably context-specific in different biological processes. While nuclear localization of βcatenin is the key to activation of the Wnt/βcatenin pathway and target gene expression, the molecular mechanisms of how the same Wnt/β-catenin signaling pathway induces specific responses remain undetermined. Recent advances in high- throughput sequencing technologies and the availability of genome information for Xenopus tropicalis have enabled us to uncover a genome-wide view of Wnt/βcatenin signaling in early vertebrate embryos, which challenges previous concepts about molecular mechanisms of Wnt target gene regulation.
    [Show full text]
  • Tsukushi Functions As a Wnt Signaling Inhibitor by Competing with Wnt2b for Binding to Transmembrane Protein Frizzled4
    Tsukushi functions as a Wnt signaling inhibitor by competing with Wnt2b for binding to transmembrane protein Frizzled4 Kunimasa Ohtaa,b,1,2, Ayako Itoa,c,1, Sei Kuriyamaa,d,3, Giuseppe Lupoe,f, Mitsuko Kosakag,4, Shin-ichi Ohnumah, Shinichi Nakagawai, and Hideaki Tanakaa,c,d aDepartment of Developmental Neurobiology, Graduate School of Medical Sciences, Kumamoto University, Kumamoto 860-8556, Japan; bPrecursory Research for Embryonic Science and Technology, Japan Science and Technology Agency, Saitama 332-0012, Japan; cGlobal Center of Excellence, Kumamoto University, Kumamoto 860-8556, Japan; d21st Century Center of Excellence, Kumamoto University, Kumamoto 860-8556, Japan; eDepartment of Biology and Biotechnology “C. Darwin,” University of Rome “La Sapienza,” 00185 Rome, Italy; fIstituto Pasteur–Fondazione Cenci Bolognetti, 00185, Rome, Italy; gRIKEN Center for Developmental Biology, Kobe 650-0047, Japan; hInstitute of Ophthalmology, University College London, London EC1V 9EL, United Kingdom; and IRIKEN Advanced Science Institute, Nakagawa RNA Biology Laboratory, Saitama 351-0198, Japan Edited* by Lynn T. Landmesser, Case Western Reserve University, Cleveland, OH, and approved July 8, 2011 (received for review January 11, 2011) The Wnt signaling pathway is essential for the development of We previously described the isolation of Tsukushi (TSK) protein diverse tissues during embryogenesis. Signal transduction is acti- isoforms (13), soluble molecules belonging to the small leucine- vated by the binding of Wnt proteins to the type I receptor low- rich proteoglycan (SLRP) family (14), and showed that they work density lipoprotein receptor–related protein 5/6 and the seven-pass as extracellular modulators of pivotal signaling cascades during transmembrane protein Frizzled (Fzd), which contains a Wnt- early embryonic development in chicks and frogs (13, 15–17).
    [Show full text]
  • Towards an Integrated View of Wnt Signaling in Development Renée Van Amerongen and Roel Nusse*
    HYPOTHESIS 3205 Development 136, 3205-3214 (2009) doi:10.1242/dev.033910 Towards an integrated view of Wnt signaling in development Renée van Amerongen and Roel Nusse* Wnt signaling is crucial for embryonic development in all animal Notably, components at virtually every level of the Wnt signal species studied to date. The interaction between Wnt proteins transduction cascade have been shown to affect both β-catenin- and cell surface receptors can result in a variety of intracellular dependent and -independent responses, depending on the cellular responses. A key remaining question is how these specific context. As we discuss below, this holds true for the Wnt proteins responses take shape in the context of a complex, multicellular themselves, as well as for their receptors and some intracellular organism. Recent studies suggest that we have to revise some of messengers. Rather than concluding that these proteins are shared our most basic ideas about Wnt signal transduction. Rather than between pathways, we instead propose that it is the total net thinking about Wnt signaling in terms of distinct, linear, cellular balance of signals that ultimately determines the response of the signaling pathways, we propose a novel view that considers the receiving cell. In the context of an intact and developing integration of multiple, often simultaneous, inputs at the level organism, cells receive multiple, dynamic, often simultaneous and of both Wnt-receptor binding and the downstream, sometimes even conflicting inputs, all of which are integrated to intracellular response. elicit the appropriate cell behavior in response. As such, the different signaling pathways might thus be more intimately Introduction intertwined than previously envisioned.
    [Show full text]
  • Hippocampus Formation: an Intriguing Collaboration Henk Roelink
    bb10g06.qxd 04/03/2000 01:10 Page R279 Dispatch R279 Hippocampus formation: An intriguing collaboration Henk Roelink Recent genetic studies have shown that the signalling temporal lobes are formed. In an adult rodent, the hip- factor Wnt3a is required for formation of the pocampus is consequently still found close to the dorsal hippocampus; the developmental consequences of Wnt midline where it is initially formed (Figure 1). signalling in the hippocampus are mediated by multiple HMG-box transcription factors, with LEF-1 being It is clear that Wnt family members are required for the required just for formation of the dentate gyrus. development of many embryonic structures, functions mediated by their effects on fundamental processes such Address: Department of Biological Structure and Center for Developmental Biology, University of Washington, Box 357420, as cell proliferation, differentiation, survival or mainte- Seattle, Washington 98117-7420, USA. nance. Obtaining a clear picture of how each Wnt acts is E-mail: [email protected] complicated by the relatively large size of the family, which has at least 18 members in amniotes [3]. Analyses of Current Biology 2000, 10:R279–R281 mice lacking one or several Wnt genes have revealed some 0960-9822/00/$ – see front matter remarkable phenotypes, often characterized by failure of © 2000 Elsevier Science Ltd. All rights reserved. induction of very specific anatomical structures. Wnt3A is no exception, and although several papers have been pub- Inductive interactions are fundamental to the formation of lished already on the more obvious defects of Wnt3a all brain structures. and signalling by molecules of the knockout mice, a recent paper from McMahon and col- Wingless/Wnt family is known to play a role in many of leagues [1] has reported a very interesting defect in the them.
    [Show full text]
  • A Membrane-Associated Β-Catenin/Oct4 Complex Correlates
    DEVELOPMENT AND STEM CELLS RESEARCH ARTICLE 1171 Development 140, 1171-1183 (2013) doi:10.1242/dev.085654 © 2013. Published by The Company of Biologists Ltd A membrane-associated β-catenin/Oct4 complex correlates with ground-state pluripotency in mouse embryonic stem cells Fernando Faunes1,*, Penelope Hayward1,*, Silvia Muñoz Descalzo1, Sujash S. Chatterjee2, Tina Balayo1, Jamie Trott1, Andrew Christoforou1,3, Anna Ferrer-Vaquer4, Anna-Katerina Hadjantonakis4, Ramanuj Dasgupta2 and Alfonso Martinez Arias1,‡ SUMMARY The maintenance of pluripotency in mouse embryonic stem cells (mESCs) relies on the activity of a transcriptional network that is fuelled by the activity of three transcription factors (Nanog, Oct4 and Sox2) and balanced by the repressive activity of Tcf3. Extracellular signals modulate the activity of the network and regulate the differentiation capacity of the cells. Wnt/β-catenin signaling has emerged as a significant potentiator of pluripotency: increases in the levels of β-catenin regulate the activity of Oct4 and Nanog, and enhance pluripotency. A recent report shows that β-catenin achieves some of these effects by modulating the activity of Tcf3, and that this effect does not require its transcriptional activation domain. Here, we show that during self-renewal there is negligible transcriptional activity of β-catenin and that this is due to its tight association with membranes, where we find it in a complex with Oct4 and E-cadherin. Differentiation triggers a burst of Wnt/β-catenin transcriptional activity that coincides with the disassembly of the complex. Our results establish that β-catenin, but not its transcriptional activity, is central to pluripotency acting through a β- catenin/Oct4 complex.
    [Show full text]
  • Beta;-Catenin Signaling in Rhabdomyosarcoma
    Laboratory Investigation (2013) 93, 1090–1099 & 2013 USCAP, Inc All rights reserved 0023-6837/13 Characterization of Wnt/b-catenin signaling in rhabdomyosarcoma Srinivas R Annavarapu1,8,9, Samantha Cialfi2,9, Carlo Dominici2,3, George K Kokai4, Stefania Uccini5, Simona Ceccarelli6, Heather P McDowell2,3,7 and Timothy R Helliwell1 Rhabdomyosarcoma (RMS) is the most common soft tissue sarcoma in children and accounts for about 5% of all malignant paediatric tumours. b-Catenin, a multifunctional nuclear transcription factor in the canonical Wnt signaling pathway, is active in myogenesis and embryonal somite patterning. Dysregulation of Wnt signaling facilitates tumour invasion and metastasis. This study characterizes Wnt/b-catenin signaling and functional activity in paediatric embryonal and alveolar RMS. Immunohistochemical assessment of paraffin-embedded tissues from 44 RMS showed b-catenin expression in 26 cases with cytoplasmic/membranous expression in 9/14 cases of alveolar RMS, and 15/30 cases of embryonal RMS, whereas nuclear expression was only seen in 2 cases of embryonal RMS. The potential functional significance of b-catenin expression was tested in four RMS cell lines, two derived from embryonal (RD and RD18) RMS and two from alveolar (Rh4 and Rh30) RMS. Western blot analysis demonstrated the expression of Wnt-associated proteins including b-catenin, glycogen synthase kinase-3b, disheveled, axin-1, naked, LRP-6 and cadherins in all cell lines. Cell fractionation and immunofluorescence studies of the cell lines (after stimulation by human recombinant Wnt3a) showed reduced phosphorylation of b-catenin, stabilization of the active cytosolic form and nuclear translocation of b-catenin. Reporter gene assay demonstrated a T-cell factor/lymphoid-enhancing factor-mediated transactivation in these cells.
    [Show full text]
  • Wnt3a: Functions and Implications in Cancer Sha He1,2, Yi Lu1,2, Xia Liu1,2, Xin Huang1,2, Evan T
    He et al. Chin J Cancer (2015) 34:50 DOI 10.1186/s40880-015-0052-4 REVIEW Open Access Wnt3a: functions and implications in cancer Sha He1,2, Yi Lu1,2, Xia Liu1,2, Xin Huang1,2, Evan T. Keller3, Chao‑Nan Qian4* and Jian Zhang1,2,3* Abstract Wnt3a, one of Wnt family members, plays key roles in regulating pleiotropic cellular functions, including self-renewal, proliferation, differentiation, and motility. Accumulating evidence has suggested that Wnt3a promotes or suppresses tumor progression via the canonical Wnt signaling pathway depending on cancer type. In addition, the roles of Wnt3a signaling can be inhibited by multiple proteins or chemicals. Herein, we summarize the latest findings on Wnt3a as an important therapeutic target in cancer. Keywords: Wnt3a, The Wnt signaling pathway, Cancer Background: the Wnt signaling pathway cell polarity pathway (Wnt-PCP pathway) and the Wnt- The Wnt gene was first identified in 1982 and is named calcium pathway (Wnt-Ca2+ pathway) [5]. According to the Int gene in mice [1]. A subsequent study reported the characteristics of its functions, the Wnt family can that the Int gene and wingless gene in Drosophila were also be divided into two categories: Wnt1 and Wnt5a. homologous genes [2]. Therefore, both genes were rec- The Wnt1 category includes Wnt1, Wnt2, Wnt2b, Wnt3, ognized as the Wnt gene [3]. The Wnt family comprises Wnt3a, Wnt7a, Wnt8, Wnt8b, and Wnt10a, which are 19 human proteins, including Wnt1, Wnt2, Wnt2b involved in the canonical signaling pathway, whereas (Wnt13), Wnt3, Wnt3a, Wnt4, Wnt5a, Wnt5b, Wnt6, Wnt4, Wnt5a, and Wnt11 belong to the Wnt5a category Wnt7a, Wnt7b, Wnt8a, Wnt8b, Wnt9a (Wnt14), Wnt9b and activate the noncanonical signaling pathway.
    [Show full text]