DOCSLIB.ORG

Pubertal Androgenization and Gonadal Histology in Two 46,XY

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Pubertal Androgenization and Gonadal Histology in Two 46,XY European Journal of Endocrinology (2012) 166 341–349 ISSN 0804-4643 CASE REPORT Pubertal androgenization and gonadal histology in two 46,XY adolescents with NR5A1 mutations and predominantly female phenotype at birth M Cools1, P Hoebeke2, K P Wolffenbuttel3, H Stoop4, R Hersmus4, M Barbaro5, A Wedell5, H Bru¨ggenwirth6, L H J Looijenga4 and S L S Drop7 1Division of Pediatric Endocrinology, Department of Pediatrics, Division of Pediatric Urology, Department of Urology, University Hospital Ghent, Ghent University, Building 3K12D, De Pintelaan 185, 9000 Ghent, Belgium, 2University Hospital Ghent, Ghent University, Ghent, Belgium, 3Division of Pediatric Urology, Department of Urology, Erasmus Medical Center, Sophia Children’s Hospital, Rotterdam, The Netherlands, 4Department of Pathology, Josephine Nefkens Institute, Daniel Den Hoed Cancer Center,Erasmus Medical Center,Rotterdam, The Netherlands, 5Department of Molecular Medicine and Surgery, Karolinska Institutet, Center for Inherited Metabolic Diseases (CMMS), Karolinska University Hospital, Stockholm, Sweden, 6Department of Clinical Genetics and 7Division of Pediatric Endocrinology, Department of Pediatrics, Erasmus Medical Center, Sophia Children’s Hospital, Rotterdam, The Netherlands (Correspondence should be addressed to M Cools; Email: [email protected]) Abstract Objective: Most patients with NR5A1 (SF-1) mutations and poor virilization at birth are sex-assigned female and receive early gonadectomy. Although studies in pituitary-specific Sf-1 knockout mice suggest hypogonadotropic hypogonadism, little is known about endocrine function at puberty and on germ cell tumor risk in patients with SF-1 mutations. This study reports on the natural course during puberty and on gonadal histology in two adolescents with SF-1 mutations and predominantly female phenotype at birth. Design and methods: Clinical and hormonal data and histopathological studies are reported in one male and one female adolescent with, respectively, a nonsense mutation (c.9TOA, p.Tyr3X) and a deletion of the first two coding exons (NCBI36/hg18 Chr9:g.(126306276-126307705)_(126303229- 126302828)del) of NR5A1, both predicted to fully disrupt gene function. Results: LH and testosterone concentrations were in the normal male range, virilization was disproportionate to the neonatal phenotype. In the girl, gonadectomy at 13 years revealed incomplete spermatogenesis and bilateral precursor lesions of testicular carcinoma in situ. In the boy, at the age of 12, numerous germ cells without signs of malignancy were present in bilateral testicular biopsy specimen. Conclusions:InSF-1 mutations, the neonatal phenotype poorly predicts virilization at puberty. Even in poorly virilized cases at birth, male gender assignment may allow spontaneous puberty without signs of hypogonadotropic hypogonadism, and possibly fertility. Patients with SF-1 mutations are at increased risk for malignant germ cell tumors. In case of preserved gonads, early orchidopexy and germ cell tumor screening is warranted. The finding of premalignant and/or malignant changes should prompt gonadectomy or possibly irradiation. European Journal of Endocrinology 166 341–349 Introduction 46,XY individuals from phenotypically females with complete gonadal dysgenesis with or without adrenal Steroidogenic factor 1 (SF-1), encoded by the NR5A1 failure (5–10) to males with penoscrotal hypospadias gene, is known to be involved in adrenal and gonadal (8, 11, 12) or bilateral anorchia (13). In 46,XX women, development and differentiation. In addition, in both NR5A1 mutations have been associated with premature males and females, SF-1 is a key promoter of gonadal ovarian failure (3). The clinical and genetic charac- hormone production, by regulating the transcription teristics of these patients have been reviewed elsewhere of several genes involved in steroidogenesis (1–3). SF-1 (14–16). mainly acts through direct binding to the promoter The observed Sf-1 expression in the ventromedial regions of its target genes, however, indirect effects have hypothalamus and gonadotrophs and impaired gonado- also been studied (1, 2, 4). trophin production in Sf-1 knockout mice suggest an The clinical presentation of patients affected additional role for Sf-1 in the normal regulation of by NR5A1 mutations is very diverse, ranging in puberty, reproduction, and metabolism (14, 17). In line q 2012 European Society of Endocrinology DOI: 10.1530/EJE-11-0392 Online version via www.eje-online.org Downloaded from Bioscientifica.com at 10/01/2021 11:34:16PM via free access 342 M Cools and others EUROPEAN JOURNAL OF ENDOCRINOLOGY (2012) 166 with this hypothesis, pituitary-specific Sf-1 knockout The PCR products were separated by capillary electro- mice develop hypogonadotropic hypogonadism (18). phoresis on an ABI 3100 Genetic Analyzer (Applied Since most of the initially reported 46,XY patients, on Biosystems, Warrington, UK). Trace data were analyzed the basis of absent or poor virilization at birth, were sex- using the Gene Mapper v3.7 software and exported to assigned female and received gonadectomy at a young an Excel spreadsheet (Microsoft). Analysis, using peak age, little is known about the natural course of puberty heights, was performed as described previously (20). in 46,XY patients with NR5A1 mutations. In addition, the risk for the development of germ cell tumors in older Sequencing DNA was extracted from peripheral blood individuals remains an important but unexplored issue. cells using standard procedures. The coding exons and In this study, we present detailed clinical, hormonal, exon/intron boundaries (exons 2–7) were amplified, and histopathological investigations at the onset of using the primers summarized in Tables 1 and 2. All puberty in two patients with NR5A1 haploinsufficiency, PCRs were performed by TaKaRa LA Taq polymerase. and with a predominantly female phenotype at birth. Subsequently, the fragments obtained were analyzed One of them was reassigned male at the age of 2 years by direct sequence analysis on an ABI 3730XL DNA after the finding of a palpable labioscrotal gonad and his sequencer. Data analysis was carried out using Seqscape testis function has been followed through puberty. software (Applied Biosystems). The sequence variant was confirmed by a second experiment, including PCR, on the same DNA sample. Material and methods Hormonal investigations Pathological studies Testosterone random values were obtained by coat-a- Gonadal tissue was fixed in 10% formalin for 24 h. Slices count RIA (Siemens, Los Angeles, CA, USA) (case 1) and of 3 mm thickness were prepared. Experimental con- liquid chromatography/tandem mass spectrometry ditions for immunohistochemistry are summarized in (UPLC Waters quattro premier, Waters Corporation, Tables 1 and 2.StainingwasperformedbyDAB/H2O2 Milford, MA, USA) (case 2). LH and FSH random values (OCT3/4 (POU5A1), SOX9, FOXL2, and SF-1 (NR5A1)) were obtained by electrochemoluminescence assay or New Fuchsin/Sodium nitrite Naphtol ASMX phosphate (Roche Diagnostics E170 Modular). Inhibin B (Inhibin (TSPYand AMH), and counterstaining with hematoxylin. B, Oxford Bio-Innovation Ltd, Oxford, UK) and AMH (Beckman Coulter Company, Suarle´e, Belgium) were Ethics measured by ELISA. Human chorionic gonadotropin (HCG) and ACTH tests were performed according to our Informed consent to publish results was obtained from local protocols. Values for testosterone were obtained at both patients and their parents. baseline and 72 h after i.m. injection of Pregnyl (Organon, Merck, NJ, USA), 1500 U. Values for cortisol were obtained at baseline and 60 min after i.v. injection Results of Synacthen (Novartis Pharma), 250 mg. Case 1 is a girl from consanguineous parents, who first came to medical attention at the age of 13 years, Gene analyses because of progressive virilization in the past year (facial hair growth, deepening of the voice, and muscular Multiplex ligation-dependent probe amplification body) and lack of breast development. Her younger (MLPA) An in-house-designed synthetic probe set for sister and brother are healthy. Discrete clitoromegaly MLPA was used to detect single exon deletions/duplica- had been noticed at birth but was judged insufficient to tions on the NR5A1 gene (19). MLPA reactions were prompt medical investigations. At presentation, she had performed by the reagents and the recommendations of pubic and axillary hair but no breast development the EK1 MLPA reagent kit (MRC-Holland, Amsterdam, (Tanner B1P3A2), clitoral length was 40 mm. On The Netherlands), starting from 100 ng genomic DNA. questionnaires on gender role behavior and gender Table 1 Summary of primers used for sequence analysis of the NR5A1 gene. Amplicon Forward primer Reverse primer NR5A1SQE0203 50-gggcacagagaggggattac-30 50-gttctcttgcagcgactgg-30 NR5A1SQE04-1 50-ggattgcctgaaaagctgag-30 50-agggctctgggtagccgta-30 NR5A1SQE04-2 50-gcttcaagctggagacagg-30 50-caaaggacagtcgggctaag-30 NR5A1SQE05 50-gaaggggtgagaggaaggtc-30 50-gtggggaaagggctgataat-30 NR5A1SQE06 50-cccatctgacctgcacct-30 50-aggatgggagcctggaataa-30 NR5A1SQE07 50-atgcccatgtctttgatggt-30 50-tcccaaacacacagtgtcaga-30 www.eje-online.org Downloaded from Bioscientifica.com at 10/01/2021 11:34:16PM via free access EUROPEAN JOURNAL OF ENDOCRINOLOGY (2012) 166 Puberty and gonadal histology in SF-1 mutations 343 Table 2 Experimental conditions for the pathological studies. Incubation Primary AB Supplied by Dilution Pretreatment AG retrieval time Secondary AB 0 OCT3/4 Santa Cruz Biotechnology, 1/1000 3% H2O2 for 5 pH 6,
Load more

Recommended publications
  • STAT3-Ser/Hes3 Signaling: a New Molecular Component of the Neuroendocrine System?
    Review 77 STAT3-Ser/Hes3 Signaling: A New Molecular Component of the Neuroendocrine System? Authors P. Nikolakopoulou1, S. W. Poser1, J. Masjkur1, M. Fernandez Rubin de Celis1, L. Toutouna1, C. L. Andoniadou2, R. D. McKay3, G. Chrousos4, M. Ehrhart-Bornstein1, S. R. Bornstein1, A. Androutsellis-Theotokis1, 5, 6 Affiliations Affiliation addresses are listed at the end of the article Key words Abstract evidence suggests that a common non-canonical ●▶ STAT3 ▼ signaling pathway regulates adult progenitors in ●▶ Hes3 The endocrine system involves communication several different tissues, rendering it as a poten- ▶ stem cells ● among different tissues in distinct organs, includ- tially valuable starting point to explore their ing the pancreas and components of the Hypo- biology. The STAT3-Ser/Hes3 Signaling Axis was thalamic-Pituitary-Adrenal Axis. The molecular first identified as a major regulator of neural mechanisms underlying these complex interac- stem cells and, subsequently, cancer stem cells. In tions are a subject of intense study as they may the endocrine/neuroendocrine system, this path- hold clues for the progression and treatment of a way operates on several levels, regulating other variety of metabolic and degenerative diseases. A types of plastic cells: (a) it regulates pancreatic plethora of signaling pathways, activated by hor- islet cell function and insulin release; (b) insulin mones and other endocrine factors have been in turn activates the pathway in broadly distrib- implicated in this communication. Recent uted neural progenitors and possibly also hypo- advances in the stem cell field introduce a new thalamic tanycytes, cells with important roles in level of complexity: adult progenitor cells appear the control of the adrenal gland; (c) adrenal pro- to utilize distinct signaling pathways than the genitors themselves operate this pathway.
    [Show full text]
  • Supplemental Table 1. Complete Gene Lists and GO Terms from Figure 3C
    Supplemental Table 1. Complete gene lists and GO terms from Figure 3C. Path 1 Genes: RP11-34P13.15, RP4-758J18.10, VWA1, CHD5, AZIN2, FOXO6, RP11-403I13.8, ARHGAP30, RGS4, LRRN2, RASSF5, SERTAD4, GJC2, RHOU, REEP1, FOXI3, SH3RF3, COL4A4, ZDHHC23, FGFR3, PPP2R2C, CTD-2031P19.4, RNF182, GRM4, PRR15, DGKI, CHMP4C, CALB1, SPAG1, KLF4, ENG, RET, GDF10, ADAMTS14, SPOCK2, MBL1P, ADAM8, LRP4-AS1, CARNS1, DGAT2, CRYAB, AP000783.1, OPCML, PLEKHG6, GDF3, EMP1, RASSF9, FAM101A, STON2, GREM1, ACTC1, CORO2B, FURIN, WFIKKN1, BAIAP3, TMC5, HS3ST4, ZFHX3, NLRP1, RASD1, CACNG4, EMILIN2, L3MBTL4, KLHL14, HMSD, RP11-849I19.1, SALL3, GADD45B, KANK3, CTC- 526N19.1, ZNF888, MMP9, BMP7, PIK3IP1, MCHR1, SYTL5, CAMK2N1, PINK1, ID3, PTPRU, MANEAL, MCOLN3, LRRC8C, NTNG1, KCNC4, RP11, 430C7.5, C1orf95, ID2-AS1, ID2, GDF7, KCNG3, RGPD8, PSD4, CCDC74B, BMPR2, KAT2B, LINC00693, ZNF654, FILIP1L, SH3TC1, CPEB2, NPFFR2, TRPC3, RP11-752L20.3, FAM198B, TLL1, CDH9, PDZD2, CHSY3, GALNT10, FOXQ1, ATXN1, ID4, COL11A2, CNR1, GTF2IP4, FZD1, PAX5, RP11-35N6.1, UNC5B, NKX1-2, FAM196A, EBF3, PRRG4, LRP4, SYT7, PLBD1, GRASP, ALX1, HIP1R, LPAR6, SLITRK6, C16orf89, RP11-491F9.1, MMP2, B3GNT9, NXPH3, TNRC6C-AS1, LDLRAD4, NOL4, SMAD7, HCN2, PDE4A, KANK2, SAMD1, EXOC3L2, IL11, EMILIN3, KCNB1, DOK5, EEF1A2, A4GALT, ADGRG2, ELF4, ABCD1 Term Count % PValue Genes regulation of pathway-restricted GDF3, SMAD7, GDF7, BMPR2, GDF10, GREM1, BMP7, LDLRAD4, SMAD protein phosphorylation 9 6.34 1.31E-08 ENG pathway-restricted SMAD protein GDF3, SMAD7, GDF7, BMPR2, GDF10, GREM1, BMP7, LDLRAD4, phosphorylation
    [Show full text]
  • HER Tyrosine Kinase Family and Rhabdomyosarcoma: Role in Onset and Targeted Therapy
    cells Review HER Tyrosine Kinase Family and Rhabdomyosarcoma: Role in Onset and Targeted Therapy Carla De Giovanni 1,* , Lorena Landuzzi 2,†, Arianna Palladini 1 , Giordano Nicoletti 2, Patrizia Nanni 1 and Pier-Luigi Lollini 1,* 1 Laboratory of Immunology and Biology of Metastasis, Department of Experimental, Diagnostic and Specialty Medicine (DIMES), Alma Mater Studiorum University of Bologna, 40126 Bologna, Italy; [email protected] (A.P.); [email protected] (P.N.) 2 Laboratory of Experimental Oncology, IRCCS Istituto Ortopedico Rizzoli, 40136 Bologna, Italy; [email protected] (L.L.); [email protected] (G.N.) * Correspondence: [email protected] (C.D.G.); [email protected] (P.-L.L.); Tel.: +39-051-2094786 (P.-L.L.) † Co-first author. Abstract: Rhabdomyosarcomas (RMS) are tumors of the skeletal muscle lineage. Two main features allow for distinction between subtypes: morphology and presence/absence of a translocation be- tween the PAX3 (or PAX7) and FOXO1 genes. The two main subtypes are fusion-positive alveolar RMS (ARMS) and fusion-negative embryonal RMS (ERMS). This review will focus on the role of receptor tyrosine kinases of the human epidermal growth factor receptor (EGFR) family that is comprised EGFR itself, HER2, HER3 and HER4 in RMS onset and the potential therapeutic targeting of receptor tyrosine kinases. EGFR is highly expressed by ERMS tumors and cell lines, in some cases contributing to tumor growth. If not mutated, HER2 is not directly involved in control of RMS cell growth but can be expressed at significant levels. A minority of ERMS carries a HER2 mutation Citation: De Giovanni, C.; Landuzzi, with driving activity on tumor growth.
    [Show full text]
  • Single Cell Derived Clonal Analysis of Human Glioblastoma Links
    SUPPLEMENTARY INFORMATION: Single cell derived clonal analysis of human glioblastoma links functional and genomic heterogeneity ! Mona Meyer*, Jüri Reimand*, Xiaoyang Lan, Renee Head, Xueming Zhu, Michelle Kushida, Jane Bayani, Jessica C. Pressey, Anath Lionel, Ian D. Clarke, Michael Cusimano, Jeremy Squire, Stephen Scherer, Mark Bernstein, Melanie A. Woodin, Gary D. Bader**, and Peter B. Dirks**! ! * These authors contributed equally to this work.! ** Correspondence: [email protected] or [email protected]! ! Supplementary information - Meyer, Reimand et al. Supplementary methods" 4" Patient samples and fluorescence activated cell sorting (FACS)! 4! Differentiation! 4! Immunocytochemistry and EdU Imaging! 4! Proliferation! 5! Western blotting ! 5! Temozolomide treatment! 5! NCI drug library screen! 6! Orthotopic injections! 6! Immunohistochemistry on tumor sections! 6! Promoter methylation of MGMT! 6! Fluorescence in situ Hybridization (FISH)! 7! SNP6 microarray analysis and genome segmentation! 7! Calling copy number alterations! 8! Mapping altered genome segments to genes! 8! Recurrently altered genes with clonal variability! 9! Global analyses of copy number alterations! 9! Phylogenetic analysis of copy number alterations! 10! Microarray analysis! 10! Gene expression differences of TMZ resistant and sensitive clones of GBM-482! 10! Reverse transcription-PCR analyses! 11! Tumor subtype analysis of TMZ-sensitive and resistant clones! 11! Pathway analysis of gene expression in the TMZ-sensitive clone of GBM-482! 11! Supplementary figures and tables" 13" "2 Supplementary information - Meyer, Reimand et al. Table S1: Individual clones from all patient tumors are tumorigenic. ! 14! Fig. S1: clonal tumorigenicity.! 15! Fig. S2: clonal heterogeneity of EGFR and PTEN expression.! 20! Fig. S3: clonal heterogeneity of proliferation.! 21! Fig.
    [Show full text]
  • Hes Genes and Neurogenin Regulate Non-Neural Versus Neural Fate
    RESEARCH ARTICLE 2531 Development 135, 2531-2541 (2008) doi:10.1242/dev.021535 Hes genes and neurogenin regulate non-neural versus neural fate specification in the dorsal telencephalic midline Itaru Imayoshi1,2,3, Tomomi Shimogori4, Toshiyuki Ohtsuka1,3 and Ryoichiro Kageyama1,3,* The choroid plexus in the brain is unique because it is a non-neural secretory tissue. It secretes the cerebrospinal fluid and functions as a blood-brain barrier, but the precise mechanism of specification of this non-neural tissue has not yet been determined. Using mouse embryos and lineage-tracing analysis, we found that the prospective choroid plexus region initially gives rise to Cajal-Retzius cells, specialized neurons that guide neuronal migration. Inactivation of the bHLH repressor genes Hes1, Hes3 and Hes5 upregulated expression of the proneural gene neurogenin 2 (Ngn2) and prematurely depleted Bmp-expressing progenitor cells, leading to enhanced formation of Cajal-Retzius cells and complete loss of choroid plexus epithelial cells. Overexpression of Ngn2 had similar effects. These data indicate that Hes genes promote specification of the fate of choroid plexus epithelial cells rather than the fate of Cajal-Retzius cells by antagonizing Ngn2 in the dorsal telencephalic midline region, and thus this study has identified a novel role for bHLH genes in the process of deciding which cells will have a non-neural versus a neural fate. KEY WORDS: Cajal-Retzius cells, Choroid plexus, Hes1, Hes5, Neurogenin, Mouse INTRODUCTION Awatramani et al., 2003; Currle et al., 2005; Hunter and Dymecki, The telencephalic hemispheres are formed by bilateral evagination 2007). The role of Bmp signaling in the development of the choroid of the anterior end of the neural tube.
    [Show full text]
  • PAX3-FOXO1 Transgenic Zebrafish Models Identify HES3 As a Mediator
    RESEARCH ARTICLE PAX3-FOXO1 transgenic zebrafish models identify HES3 as a mediator of rhabdomyosarcoma tumorigenesis Genevieve C Kendall1,2, Sarah Watson3, Lin Xu1, Collette A LaVigne1,2, Whitney Murchison1, Dinesh Rakheja1,4, Stephen X Skapek1, Franck Tirode5, Olivier Delattre3,6,7, James F Amatruda1,2,8* 1Department of Pediatrics, UT Southwestern Medical Center, Dallas, United States; 2Department of Molecular Biology, UT Southwestern Medical Center, Dallas, United States; 3Institut Curie, Paris Sciences et Lettres (PSL) Research University, Inserm U830, Institut Curie, Paris Sciences et Lettres (PSL) Research University, Paris, France; 4Department of Pathology, UT Southwestern Medical Center, Dallas, United States; 5Univ Lyon, Universite´ Claude Bernard Lyon 1, INSERM 1052, CNRS 5286, Centre Le´onBe´rard, Lyon, France; 6INSERM U80, Institute Curie Research Center, Paris, France; 7Institut Curie Hospital Group, Unite´ de Ge´ne´tique Somatique, Paris, France; 8Department of Internal Medicine, UT Southwestern Medical Center, Dallas, United States Abstract Alveolar rhabdomyosarcoma is a pediatric soft-tissue sarcoma caused by PAX3/7- FOXO1 fusion oncogenes and is characterized by impaired skeletal muscle development. We developed human PAX3-FOXO1 -driven zebrafish models of tumorigenesis and found that PAX3- FOXO1 exhibits discrete cell lineage susceptibility and transformation. Tumors developed by 1.6– 19 months and were primitive neuroectodermal tumors or rhabdomyosarcoma. We applied this PAX3-FOXO1 transgenic zebrafish model to study how PAX3-FOXO1 leverages early *For correspondence: developmental pathways for oncogenesis and found that her3 is a unique target. Ectopic james.amatruda@utsouthwestern. expression of the her3 human ortholog, HES3, inhibits myogenesis in zebrafish and mammalian edu cells, recapitulating the arrested muscle development characteristic of rhabdomyosarcoma.
    [Show full text]
  • Human Embryonic Stem Cells Facilitate Isolation of in Vitro Derived Insulin-Producing Cells
    Diabetologia (2012) 55:694–706 DOI 10.1007/s00125-011-2379-y ARTICLE INSGFP/w human embryonic stem cells facilitate isolation of in vitro derived insulin-producing cells S. J. Micallef & X. Li & J. V. Schiesser & C. E. Hirst & Q. C. Yu & S. M. Lim & M. C. Nostro & D. A. Elliott & F. Sarangi & L. C. Harrison & G. Keller & A. G. Elefanty & E. G. Stanley Received: 27 July 2011 /Accepted: 20 October 2011 /Published online: 26 November 2011 # The Author(s) 2011. This article is published with open access at Springerlink.com Abstract spin embryoid body (EB) differentiation protocol that used Aims/hypothesis We aimed to generate human embryonic the recombinant protein-based, fully defined medium, stem cell (hESC) reporter lines that would facilitate the APEL. Like INS-GFP+ cells generated with other methods, characterisation of insulin-producing (INS+) cells derived those derived using the spin EB protocol expressed a suite in vitro. of pancreatic-related transcription factor genes including Methods Homologous recombination was used to insert ISL1, PAX6 and NKX2.2. However, in contrast with sequences encoding green fluorescent protein (GFP) into previous methods, the spin EB protocol yielded INS-GFP+ the INS locus, to create reporter cell lines enabling the cells that also co-expressed the beta cell transcription factor prospective isolation of viable INS+ cells. gene, NKX6.1, and comprised a substantial proportion of GFP/w Results Differentiation of INS hESCs using published monohormonal INS+ cells. protocols demonstrated that all GFP+ cells co-produced Conclusions/interpretation INSGFP/w hESCs are a valuable insulin, confirming the fidelity of the reporter gene.
    [Show full text]
  • Coordinated Waves of Gene Expression During Neuronal Differentiation of Embryonic Stem Cells As Basis for Novel Approaches to Developmental Neurotoxicity Testing
    Cell Death and Differentiation (2011) 18, 383–395 & 2011 Macmillan Publishers Limited All rights reserved 1350-9047/11 www.nature.com/cdd Coordinated waves of gene expression during neuronal differentiation of embryonic stem cells as basis for novel approaches to developmental neurotoxicity testing B Zimmer1, PB Kuegler1, B Baudis1, A Genewsky1, V Tanavde2, W Koh2, B Tan2, T Waldmann1, S Kadereit1 and M Leist*,1 As neuronal differentiation of embryonic stem cells (ESCs) recapitulates embryonic neurogenesis, disturbances of this process may model developmental neurotoxicity (DNT). To identify the relevant steps of in vitro neurodevelopment, we implemented a differentiation protocol yielding neurons with desired electrophysiological properties. Results from focussed transcriptional profiling suggested that detection of non-cytotoxic developmental disturbances triggered by toxicants such as retinoic acid (RA) or cyclopamine was possible. Therefore, a broad transcriptional profile of the 20-day differentiation process was obtained. Cluster analysis of expression kinetics, and bioinformatic identification of overrepresented gene ontologies revealed waves of regulation relevant for DNT testing. We further explored the concept of superimposed waves as descriptor of ordered, but overlapping biological processes. The initial wave of transcripts indicated reorganization of chromatin and epigenetic changes. Then, a transient upregulation of genes involved in the formation and patterning of neuronal precursors followed. Simultaneously, a long wave of ongoing neuronal differentiation started. This was again superseded towards the end of the process by shorter waves of neuronal maturation that yielded information on specification, extracellular matrix formation, disease-associated genes and the generation of glia. Short exposure to lead during the final differentiation phase, disturbed neuronal maturation.
    [Show full text]
  • Examining New Interactors of Notch/Rbpjκ Signalling in Adult
    TECHNISCHE UNIVERSITÄT MÜNCHEN Lehrstuhl Entwicklungsgenetik Examining new interactors of Notch/RBPJ signalling in adult neural stem cells Dissertation von Tobias Johannes Schwarz Helmholtz Zentrum München Institut für Entwicklungsgenetik TECHNISCHE UNIVERSITÄT MÜNCHEN Lehrstuhl Entwicklungsgenetik Examining new interactors of Notch/RBPJ signalling in adult neural stem cells Tobias Johannes Schwarz Vollständiger Abdruck der von der Fakultät Wissenschaftszentrum Weihenstephan für Ernährung, Landnutzung und Umwelt der Technischen Universität München zur Erlangung des akademischen Grades eines Doktors der Naturwissenschaften genehmigten Dissertation. Vorsitzende: Univ.-Prof. A. Schnieke, Ph.D. Prüfer der Dissertation: 1. Univ.-Prof. Dr. W. Wurst 2. Univ.-Prof. Dr. C. Lie, (Friedrich-Alexander Universität Erlangen-Nürnberg) Die Dissertation wurde am 20. 03. 2012 bei der Technischen Universität München eingereicht und durch die Fakultät Wissenschaftszentrum Weihenstephan für Ernährung, Landnutzung und Umwelt am 25. 07. 2012 angenommen. Table of contents 1 Zusammenfassung......................................................................................... 4 2 Summary .......................................................................................................... 6 3 Introduction ...................................................................................................... 7 3.1 Adult neurogenesis and neural stem cells ..................................................... 7 3.2 Signal integration by adult neural stem cells
    [Show full text]
  • Non-Classical 1P36 Deletion in a Patient
    Yokoyama et al. Mol Cytogenet (2020) 13:42 https://doi.org/10.1186/s13039-020-00510-5 CASE REPORT Open Access Non-classical 1p36 deletion in a patient with Duane retraction syndrome: case report and literature review Emiy Yokoyama1, Camilo E. Villarroel1, Sinhué Diaz2, Victoria Del Castillo1, Patricia Pérez‑Vera3, Consuelo Salas3, Samuel Gómez4, Reneé Barreda1, Bertha Molina5 and Sara Frias5,6* Abstract Background: Monosomy of 1p36 is considered the most common terminal microdeletion syndrome. It is character‑ ized by intellectual disability, growth retardation, seizures, congenital anomalies, and distinctive facial features that are absent when the deletion is proximal, beyond the 1p36.32 region. In patients with proximal deletions, little is known about the associated phenotype, since only a few cases have been reported in the literature. Ocular manifestations in patients with classical 1p36 monosomy are frequent and include strabismus, myopia, hypermetropia, and nystagmus. However, as of today only one patient with 1p36 deletion and Duane retraction syndrome (DRS) has been reported. Case presentation: We describe a patient with intellectual disability, facial dysmorphism, and bilateral Duane retrac‑ tion syndrome (DRS) type 1. Array CGH showed a 7.2 Mb de novo deletion from 1p36.31 to 1p36.21. Discussion: Our patient displayed DRS, which is not part of the classical phenotype and is not a common clinical feature in 1p36 deletion syndrome; we hypothesized that this could be associated with the overlapping deletion between the distal and proximal 1p36 regions. DRS is one of the Congenital Cranial Dysinnervation Disorders, and a genetic basis for the syndrome has been extensively reported.
    [Show full text]
  • JJP Suppl 2006.Fm
    Lectures Lectures L1 (2PA) L3 (1EA) The ins and outs of glutamate receptor Water and ion channels in kidney function trafficking during synaptic plasticity Sasaki, Sei (Dept. Nephrology. Grad. Sch.Tokyo Med Dent Univ. Nicoll, Roger A. (Department of Cellular and Molecular Tokyo, Japan) Pharmacology, University of California at San Francisco, San Kidney is the main organ in the maintenance of water and electrolyte ho- Francisco, California, USA) meostasis of extracellular fluid of the body. Extensive physiological re- Glutamate, the major excitatory neurotransmitter in the brain, acts pri- search has been performed to understand this important kidney function. marily on two types of ionotropic receptors, AMPA receptors and The research has developed from the organ level to molecular level and NMDA receptors. Work over the past decade indicates that the number the development was accompanied by advancement of experimental of synaptic AMPA receptors is tightly regulated and may serve as a technology, i.e., from the clearance method to molecular biology. Molec- mechanism for information storage. Recent studies show that stargazin, ular biological studies have identified a wide variety of channel and the mutated protein in the ataxic and epileptic mouse stargazer, is neces- transporter proteins that exist in renal epithelial cells. Coupled with hu- sary for the expression of surface AMPA receptors in cerebellar granule man genetic studies it has been shown that mutations of the genes encod- cells. Stargazin is a small tetraspanning membrae protein and is a mem- ing renal membrane transport proteins (channels and transporters) cause ber of a family of proteins referred to as transmembrane AMPAR regu- human hereditary diseases such as water and solute loosing or retaining latory proteins (TARPs).
    [Show full text]
  • Characterization of CSL Complexes in the Notch Pathway: the Su(H)-NICD Interaction and the RBP-J-DNA Interaction
    Characterization of CSL Complexes in the Notch Pathway: the Su(H)-NICD Interaction and the RBP-J-DNA Interaction Ashley N. Contreras B.A., Earlham College, 2007 Committee Chair: Rhett A. Kovall, Ph.D. A dissertation submitted to the Division of Graduate Studies and Research of the University of Cincinnati In partial fulfillment of the requirements for the degree of Doctor of Philosophy (Ph.D.) In the Department of Molecular Genetics, Biochemistry, and Microbiology of the College of Medicine November 2014 Abstract In metazoans, the Notch pathway is a conserved cell-to-cell signaling mechanism that plays a key role in cellular specification. It is essential for cell fate determination in embryonic development, organogenesis, hematopoiesis, and adult tissue homeostasis. Aberrant Notch signaling has been implicated in various cancers, developmental defects, and cardiovascular disease. Despite these various roles, Notch signaling is relatively simple; extracellular ligand-receptor binding triggers proteolytic processing of the Notch receptor. The newly generated receptor fragment, the Notch intracellular domain (NICD), translocates to the nucleus and interacts with the DNA-binding protein CSL (CBF1/RBPJ in mammals; Suppressor of Hairless [Su(H)] in flies; Lag-1 in worms), to ultimately activate transcription of Notch responsive genes. In the absence of pathway activation, however, CSL also represses transcription of these genes. The ability of CSL to differentially regulate gene expression is determined by its interaction with co-regulators (co-activators or co-repressors). The research described in the following chapters characterizes the molecular details of two different interactions of CSL—the Su(H)-NICD interaction and the RBP-J-DNA interaction.
    [Show full text]