DOCSLIB.ORG

Pax6 During Visual System Development

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Pax6 During Visual System Development Hedgehog-dependent E3-ligase Midline1 regulates ubiquitin-mediated proteasomal degradation of Pax6 during visual system development Thorsten Pfirrmanna,1, Enrico Jandta,1, Swantje Ranfta,b, Ashwin Lokapallya, Herbert Neuhausa, Muriel Perronc, and Thomas Hollemanna,2 aInstitute for Physiological Chemistry, University of Halle-Wittenberg, 06114 Halle, Germany; bGynecological Hospital, University Medical Center Mannheim, 68167 Mannheim, Germany; and cParis-Saclay Institute of Neuroscience, CNRS, Univ Paris Sud, Université Paris-Saclay, 91405 Orsay, France Edited by Richard M. Harland, University of California, Berkeley, CA, and approved July 19, 2016 (received for review January 16, 2016) Pax6 is a key transcription factor involved in eye, brain, and pancreas remains unclear how Pax6 protein is removed from the eyestalk development. Although pax6 is expressed in the whole prospective territory on time. Some authors report the regulation of Pax6 retinal field, subsequently its expression becomes restricted to the activity by posttranslational modifications (21–23), and most optic cup by reciprocal transcriptional repression of pax6 and pax2. interestingly, Tuoc et al. showed that in cortical progenitor cells, However, it remains unclear how Pax6 protein is removed from the Pax6 protein is degraded by the proteasome mediated by Trim11 eyestalk territory on time. Here, we report that Mid1, a member of (24). However, the existence of similar mechanisms leading to the RBCC/TRIM E3 ligase family, which was first identified in patients the development of the visual system is not known. with the X-chromosome–linked Opitz BBB/G (OS) syndrome, inter- The data of our present study show that Midline1 (Mid1) acts with Pax6. We found that the forming eyestalk is a major do- serves as one of these links. MID1, a member of the RBCC/ main of mid1 expression, controlled by the morphogen Sonic TRIM E3 ligase family, was first identified in patients with the hedgehog (Shh). Here, Mid1 regulates the ubiquitination and protea- X-chromosome–linked Opitz BBB/G (OS) syndrome. Patients somal degradation of Pax6 protein. Accordantly, when Mid1 levels suffer from multiple malformations of the ventral midline as a are knocked down, Pax6 expression is expanded and eyes are enlarged. consequence of mutations in the MID1 gene (25, 26). In previous Our findings indicate that remaining or misaddressed Pax6 protein is studies, Mid1 has been described to regulate protein phospha- cleared from the eyestalk region to properly set the border between tase 2A (PP2A) stability (27–30). PP2A/α4 is not the only known the eyestalk territory and the retina via Mid1. Thus, we identified a substrate, because recently it was shown that Mid1 interacts with posttranslational mechanism, regulated by Sonic hedgehog, which is the GLI regulator Fu, leading to a cytoplasmic retention of GLI3 important to suppress Pax6 activity and thus breaks pax6 autoregu- in cancer cells (31). We show that Mid1 can physically interact lation at defined steps during the formation of the visual system. with Pax6 leading to the ubiquitination and proteasomal degra- dation of Pax6 protein. We observe an overlapping expression of Xenopus | mid1 | pax6 | eye development | ubiquitin mid1 and pax6 in early stages of Xenopus development. In tad- pole stages, mid1 transcripts are concentrated in the optic stalk ax6 is an evolutionary highly conserved transcription factor, territory in contrast to pax6. Overexpression of shh strongly in- Pplaying key roles as a potent cell fate determinant in the duces mid1, whose expression expands to the whole remaining development of the eye, brain, and pancreas (1–4). Pax6 is a eye vesicle. Accordingly, when Mid1 levels are knocked down, Pax6 member of the PAX family of transcription factors (5, 6). Pax6 expression is expanded, and eyes are enlarged. Taken together, we autoregulation was suggested based on mouse genetic experi- ments (7) and studies regarding the quail, as well as human, Significance promoters (8, 9). Thus, to maintain pax6 expression, Pax6 pro- tein is required, which has been assumed already from studies of The first morphological sign of vertebrate eye formation is the the Small eye (SEY) mutant mice (10, 11). In humans, hetero- appearance of eye vesicles on both sides of the ventral di- zygous mutations in PAX6 cause a wide variety of ocular defects, encephalon, which primarily originate from a uniform Pax6- whereas the homozygous loss results in anophthalmia (12). Not positive eye field. A key regulator for this process, which co- only does the reduction of Pax6 protein levels cause severe de- incides with the establishment of the proximo-distal axis of the velopmental defects in the eye, but transgenic mice carrying eye with eyestalk and retina, is the morphogen Shh. Shh in- multiple copies of the human PAX6 gene also have similar ocular duces the expression of pax2, which represses pax6 in the abnormalities as the small eye mice (13, 14). Moreover, in presumptive eyestalk region. Here, we report on the identifi- Drosophila and in vertebrates like Xenopus, overexpression of cation of the E3-ligase Mid1, which is induced by Shh and pax6 is able to induce the formation of ectopic eyes (15, 16). As a targets transcription factor Pax6 for proteasomal degradation. whole, Pax6 is one of the most important regulators of eye de- Our findings might provide insight into a mechanism, in which velopment and its function is critically dependent on a tempo- a morphogen initiates the degradation of a transcription factor rarily and quantitatively defined expression level (4). to form sharp boundaries of gene expression. In Xenopus, pax6 can be detected in all neuroblasts of the BIOLOGY developing retina soon after gastrulation. In early tailbud stages, Author contributions: M.P. and T.H. designed research; T.P., E.J., S.R., A.L., H.N., M.P., and DEVELOPMENTAL pax6 is expressed homogenously in all parts of the optic vesicle. T.H. performed research; T.P., E.J., and H.N. contributed new reagents/analytic tools; T.H. In tadpoles, pax6 is limited to the lens epithelium and later to analyzed data; and T.H. wrote the paper. cells of the inner nuclear layer and the ganglion cell layer. The authors declare no conflict of interest. However, pax6 transcripts are barely detectable in the outer This article is a PNAS Direct Submission. nuclear layer containing photoreceptor cells but clearly visible in 1T.P. and E.J. contributed equally to this work. the ciliary marginal zone (17, 18). On the one hand, the sharp 2To whom correspondence should be addressed. Email: [email protected] boundary between the optic cup with pax6 and the optic stalk halle.de. region with pax2 is established by reciprocal transcriptional re- This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10. pression of these two pax genes (19, 20). On the other hand, it 1073/pnas.1600770113/-/DCSupplemental. www.pnas.org/cgi/doi/10.1073/pnas.1600770113 PNAS | September 6, 2016 | vol. 113 | no. 36 | 10103–10108 Downloaded by guest on September 23, 2021 provide evidence that Pax6 is a substrate for Mid1 and insights into single cell of two-cell stage embryos and examined mid1 ex- how remaining or misaddressed Pax6 protein is cleared from the pression at tadpole stage. Misexpression of shh resulted in an up- eyestalk region to properly set the border between the eyestalk regulation of the known target, the hh receptor ptc1 (Fig. 1 B, d–d′ territory and the retina at the right time. and Fig. S1B,nptc1= 20/27; nctrl = 6/60; P = 1.42E-11) in neural ectoderm. Moreover, mid1 expression was strongly induced but Results restricted to the optic cup (Fig. 1 B, a–a′;nmid1= 66/81; nctrl = Mid1 Expression in Cells of the Forming Optic Stalk Is Under the 3/44; P = 1.08E-15) similar to pax2 (Fig. 1 B, b–b′;npax2= 74/83; Control of Shh. We analyzed the expression of mid1 during Xen- nctrl = 3/42; P = 4.21E-18), whereas pax6 was repressed on shh opus laevis development by whole mount in situ hybridizations RNA injection (Fig. 1 B, c–c′;npax6= 86/106; nctrl = 4/51; P = (Wmish) on embryos of Nieuwkoop and Faber (NF) stage 24–38 3.47E-18) (33). Interestingly, Hh-dependent regulation of mid1 is (Fig. 1A). While confirming the pattern described by Suzuki not mediated by pax2 because suppression of pax2 function did not – ′ et al. (32), a closer look revealed that mid1 is expressed early in impair mid1 expression (Fig. 1 B, e e ; 8/11 eyes analyzed in sec- tion) but resulted in an enlargement of the retina and ectopic pax6 the whole eye vesicle. From the NF stage 32 onward, high levels – ′ of mid1 transcripts were detected in the ventral part of the expression in the eye vesicle (Fig. 1B, f f ; 13/16 eyes analyzed in pax2 pax6 forming eye and especially within the forming optic stalk (Fig. 1 section). Moreover, is under the control of ,because suppression of pax6 ledtoanexpansionofpax2 expression into the A, c4–c7 and d1–d3). Here, the expression of mid1 appears remaining eye vesicle (Fig. 1 B, g–g″; npax2 = 7/17; nctrl = 4/47; P = similar, although not identical to those described for transcrip- 1.45E-08) and pax2 RNA injection impaired pax6 expression as in tion factors vax1 or pax2, known targets of the hedgehog (hh) mice (Fig. 1 B, h–h″ and Fig. S1C; npax6 = 9/35; nctrl = 4/47; P = pathway (Fig. S1A) (33). To analyze whether Hedgehog also 1.5E-14) (20). Because Mid1 is an E3 ubiquitin ligase, we assume regulates mid1 expression, we injected synthetic shh RNA into a that Mid1 regulates Pax6 expression on a posttranslational level rather than on a transcriptional level like Pax2. Mid1 Interacts with Pax6.
Load more

Recommended publications
  • Transcriptional Regulation by Extracellular Signals 209
    Cell, Vol. 80, 199-211, January 27, 1995, Copyright © 1995 by Cell Press Transcriptional Regulation Review by Extracellular Signals: Mechanisms and Specificity Caroline S. Hill and Richard Treisman Nuclear Translocation Transcription Laboratory In principle, regulated nuclear localization of transcription Imperial Cancer Research Fund factors can involve regulated activity of either nuclear lo- Lincoln's Inn Fields calization signals (NLSs) or cytoplasmic retention signals, London WC2A 3PX although no well-characterized case of the latter has yet England been reported. N LS activity, which is generally dependent on short regions of basic amino acids, can be regulated either by masking mechanisms or by phosphorylations Changes in cell behavior induced by extracellular signal- within the NLS itself (Hunter and Karin, 1992). For exam- ing molecules such as growth factors and cytokines re- ple, association with an inhibitory subunit masks the NLS quire execution of a complex program of transcriptional of NF-KB and its relatives (Figure 1; for review see Beg events. While the route followed by the intracellular signal and Baldwin, 1993), while an intramolecular mechanism from the cell membrane to its transcription factor targets may mask NLS activity in the heat shock regulatory factor can be traced in an increasing number of cases, how the HSF2 (Sheldon and Kingston, 1993). When transcription specificity of the transcriptional response of the cell to factor localization is dependent on regulated NLS activity, different stimuli is determined is much less clear. How- linkage to a constitutively acting NLS may be sufficient to ever, it is possible to understand at least in principle how render nuclear localization independent of signaling (Beg different stimuli can activate the same signal pathway yet et al., 1992).
    [Show full text]
  • Homeobox Gene Expression Profile in Human Hematopoietic Multipotent
    Leukemia (2003) 17, 1157–1163 & 2003 Nature Publishing Group All rights reserved 0887-6924/03 $25.00 www.nature.com/leu Homeobox gene expression profile in human hematopoietic multipotent stem cells and T-cell progenitors: implications for human T-cell development T Taghon1, K Thys1, M De Smedt1, F Weerkamp2, FJT Staal2, J Plum1 and G Leclercq1 1Department of Clinical Chemistry, Microbiology and Immunology, Ghent University Hospital, Ghent, Belgium; and 2Department of Immunology, Erasmus Medical Center, Rotterdam, The Netherlands Class I homeobox (HOX) genes comprise a large family of implicated in this transformation proces.14 The HOX-C locus transcription factors that have been implicated in normal and has been primarily implicated in lymphomas.15 malignant hematopoiesis. However, data on their expression or function during T-cell development is limited. Using degener- Hematopoietic cells are derived from stem cells that reside in ated RT-PCR and Affymetrix microarray analysis, we analyzed fetal liver (FL) in the embryo and in the adult bone marrow the expression pattern of this gene family in human multipotent (ABM), which have the unique ability to self-renew and thereby stem cells from fetal liver (FL) and adult bone marrow (ABM), provide a life-long supply of blood cells. T lymphocytes are a and in T-cell progenitors from child thymus. We show that FL specific type of hematopoietic cells that play a major role in the and ABM stem cells are similar in terms of HOX gene immune system. They develop through a well-defined order of expression, but significant differences were observed between differentiation steps in the thymus.16 Several transcription these two cell types and child thymocytes.
    [Show full text]
  • Prox1regulates the Subtype-Specific Development of Caudal Ganglionic
    The Journal of Neuroscience, September 16, 2015 • 35(37):12869–12889 • 12869 Development/Plasticity/Repair Prox1 Regulates the Subtype-Specific Development of Caudal Ganglionic Eminence-Derived GABAergic Cortical Interneurons X Goichi Miyoshi,1 Allison Young,1 Timothy Petros,1 Theofanis Karayannis,1 Melissa McKenzie Chang,1 Alfonso Lavado,2 Tomohiko Iwano,3 Miho Nakajima,4 Hiroki Taniguchi,5 Z. Josh Huang,5 XNathaniel Heintz,4 Guillermo Oliver,2 Fumio Matsuzaki,3 Robert P. Machold,1 and Gord Fishell1 1Department of Neuroscience and Physiology, NYU Neuroscience Institute, Smilow Research Center, New York University School of Medicine, New York, New York 10016, 2Department of Genetics & Tumor Cell Biology, St. Jude Children’s Research Hospital, Memphis, Tennessee 38105, 3Laboratory for Cell Asymmetry, RIKEN Center for Developmental Biology, Kobe 650-0047, Japan, 4Laboratory of Molecular Biology, Howard Hughes Medical Institute, GENSAT Project, The Rockefeller University, New York, New York 10065, and 5Cold Spring Harbor Laboratory, Cold Spring Harbor, New York 11724 Neurogliaform (RELNϩ) and bipolar (VIPϩ) GABAergic interneurons of the mammalian cerebral cortex provide critical inhibition locally within the superficial layers. While these subtypes are known to originate from the embryonic caudal ganglionic eminence (CGE), the specific genetic programs that direct their positioning, maturation, and integration into the cortical network have not been eluci- dated. Here, we report that in mice expression of the transcription factor Prox1 is selectively maintained in postmitotic CGE-derived cortical interneuron precursors and that loss of Prox1 impairs the integration of these cells into superficial layers. Moreover, Prox1 differentially regulates the postnatal maturation of each specific subtype originating from the CGE (RELN, Calb2/VIP, and VIP).
    [Show full text]
  • Cell Reprogramming Technologies for Treatment And
    CELL REPROGRAMMING TECHNOLOGIES FOR TREATMENT AND UNDERSTANDING OF GENETIC DISORDERS OF MYELIN by ANGELA MARIE LAGER Submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy Thesis advisor: Paul J Tesar, PhD Department of Genetics and Genome Sciences CASE WESTERN RESERVE UNIVERSITY May 2015 CASE WESTERN RESERVE UNIVERSITY SCHOOL OF GRADUATE STUDIES We hereby approve the thesis/dissertation of Angela Marie Lager Candidate for the Doctor of Philosophy degree*. (signed) Ronald A Conlon, PhD (Committee Chair) Paul J Tesar, PhD (Advisor) Craig A Hodges, PhD Warren J Alilain, PhD (date) 31 March 2015 *We also certify that written approval has been obtained from any proprietary material contained therein. TABLE OF CONTENTS Table of Contents……………………………………………………………………….1 List of Figures……………………………………………………………………………4 Acknowledgements……………………………………………………………………..7 Abstract…………………………………………………………………………………..8 Chapter 1: Introduction and Background………………………………………..11 1.1 Overview of mammalian oligodendrocyte development in the spinal cord and myelination of the central nervous system…………………..11 1.1.1 Introduction……………………………………………………..11 1.1.2 The establishment of the neuroectoderm and ventral formation of the neural tube…………………………………..12 1.1.3 Ventral patterning of the neural tube and specification of the pMN domain in the spinal cord……………………………….15 1.1.4 Oligodendrocyte progenitor cell production through the process of gliogenesis ………………………………………..16 1.1.5 Oligodendrocyte progenitor cell to oligodendrocyte differentiation…………………………………………………...22
    [Show full text]
  • Viewed in [2, 3])
    Yildiz et al. Neural Development (2019) 14:5 https://doi.org/10.1186/s13064-019-0129-x RESEARCH ARTICLE Open Access Zebrafish prdm12b acts independently of nkx6.1 repression to promote eng1b expression in the neural tube p1 domain Ozge Yildiz1, Gerald B. Downes2 and Charles G. Sagerström1* Abstract Background: Functioning of the adult nervous system depends on the establishment of neural circuits during embryogenesis. In vertebrates, neurons that make up motor circuits form in distinct domains along the dorsoventral axis of the neural tube. Each domain is characterized by a unique combination of transcription factors (TFs) that promote a specific fate, while repressing fates of adjacent domains. The prdm12 TF is required for the expression of eng1b and the generation of V1 interneurons in the p1 domain, but the details of its function remain unclear. Methods: We used CRISPR/Cas9 to generate the first germline mutants for prdm12 and employed this resource, together with classical luciferase reporter assays and co-immunoprecipitation experiments, to study prdm12b function in zebrafish. We also generated germline mutants for bhlhe22 and nkx6.1 to examine how these TFs act with prdm12b to control p1 formation. Results: We find that prdm12b mutants lack eng1b expression in the p1 domain and also possess an abnormal touch-evoked escape response. Using luciferase reporter assays, we demonstrate that Prdm12b acts as a transcriptional repressor. We also show that the Bhlhe22 TF binds via the Prdm12b zinc finger domain to form a complex. However, bhlhe22 mutants display normal eng1b expression in the p1 domain. While prdm12 has been proposed to promote p1 fates by repressing expression of the nkx6.1 TF, we do not observe an expansion of the nkx6.1 domain upon loss of prdm12b function, nor is eng1b expression restored upon simultaneous loss of prdm12b and nkx6.1.
    [Show full text]
  • CNS Myelin Paranodes Require Nkx6-2 Homeoprotein Transcriptional Activity for Normal Structure
    The Journal of Neuroscience, December 15, 2004 • 24(50):11215–11225 • 11215 Development/Plasticity/Repair CNS Myelin Paranodes Require Nkx6-2 Homeoprotein Transcriptional Activity for Normal Structure Cherie Southwood,1,2 Chris He,1 James Garbern,2,4 John Kamholz,2,4 Edgardo Arroyo,5 and Alexander Gow1,2,3,4 1Brookdale Center for Molecular Biology, Mount Sinai School of Medicine, New York, New York, 10029, 2Center for Molecular Medicine and Genetics, 3Carman and Ann Adams Department of Pediatrics, 4Department of Neurology, Wayne State University School of Medicine, Detroit, Michigan, 48201, and 5Department of Neurology, University of Pennsylvania Medical Center, Philadelphia, Pennsylvania, 19104 Homeodomain proteins play critical roles during development in cell fate determination and proliferation, but few studies have defined gene regulatory networks for this class of transcription factors in differentiated cells. Using a lacZ-knock-in strategy to ablate Nkx6-2, we find that the Nkx6-2 promoter is active embryonically in neuroblasts and postnatally in oligodendrocytes. In addition to neurological deficits, we find widespread ultrastructural abnormalities in CNS white matter and aberrant expression of three genes encoding a paranodal microtubule destabilizing protein, stathmin 1, and the paranodal cell adhesion molecules neurofascin and contactin. The involvement of these downstream proteins in cytoskeletal function and cell adhesion reveals mechanisms whereby Nkx6-2 directly or indirectly regulates axon–glial interactions at myelin paranodes. Nkx6-2 does not appear to be the central regulator of axoglial junction assembly; nonetheless, our data constitute the first evidence of such a regulatory network and provide novel insights into the mechanism and effector molecules that are involved.
    [Show full text]
  • Genome-Wide DNA Methylation Analysis of KRAS Mutant Cell Lines Ben Yi Tew1,5, Joel K
    www.nature.com/scientificreports OPEN Genome-wide DNA methylation analysis of KRAS mutant cell lines Ben Yi Tew1,5, Joel K. Durand2,5, Kirsten L. Bryant2, Tikvah K. Hayes2, Sen Peng3, Nhan L. Tran4, Gerald C. Gooden1, David N. Buckley1, Channing J. Der2, Albert S. Baldwin2 ✉ & Bodour Salhia1 ✉ Oncogenic RAS mutations are associated with DNA methylation changes that alter gene expression to drive cancer. Recent studies suggest that DNA methylation changes may be stochastic in nature, while other groups propose distinct signaling pathways responsible for aberrant methylation. Better understanding of DNA methylation events associated with oncogenic KRAS expression could enhance therapeutic approaches. Here we analyzed the basal CpG methylation of 11 KRAS-mutant and dependent pancreatic cancer cell lines and observed strikingly similar methylation patterns. KRAS knockdown resulted in unique methylation changes with limited overlap between each cell line. In KRAS-mutant Pa16C pancreatic cancer cells, while KRAS knockdown resulted in over 8,000 diferentially methylated (DM) CpGs, treatment with the ERK1/2-selective inhibitor SCH772984 showed less than 40 DM CpGs, suggesting that ERK is not a broadly active driver of KRAS-associated DNA methylation. KRAS G12V overexpression in an isogenic lung model reveals >50,600 DM CpGs compared to non-transformed controls. In lung and pancreatic cells, gene ontology analyses of DM promoters show an enrichment for genes involved in diferentiation and development. Taken all together, KRAS-mediated DNA methylation are stochastic and independent of canonical downstream efector signaling. These epigenetically altered genes associated with KRAS expression could represent potential therapeutic targets in KRAS-driven cancer. Activating KRAS mutations can be found in nearly 25 percent of all cancers1.
    [Show full text]
  • Expanding the Phenotypic Spectrum of PAX6 Mutations: from Congenital Cataracts to Nystagmus
    G C A T T A C G G C A T genes Article Expanding the Phenotypic Spectrum of PAX6 Mutations: From Congenital Cataracts to Nystagmus Maria Nieves-Moreno 1,* , Susana Noval 1 , Jesus Peralta 1, María Palomares-Bralo 2 , Angela del Pozo 3 , Sixto Garcia-Miñaur 4, Fernando Santos-Simarro 4 and Elena Vallespin 5 1 Department of Ophthalmology, Hospital Universitario La Paz, 28046 Madrid, Spain; [email protected] (S.N.); [email protected] (J.P.) 2 Department of Molecular Developmental Disorders, Medical and Molecular Genetics Institue (INGEMM) IdiPaz, CIBERER, Hospital Universitario La Paz, 28046 Madrid, Spain; [email protected] 3 Department of Bioinformatics, Medical and Molecular Genetics Institue (INGEMM) IdiPaz, CIBERER, Hospital Universitario La Paz, 28046 Madrid, Spain; [email protected] 4 Department of Clinical Genetics, Medical and Molecular Genetics Institue (INGEMM) IdiPaz, CIBERER, Hospital Universitario La Paz, 28046 Madrid, Spain; [email protected] (S.G.-M.); [email protected] (F.S.-S.) 5 Department of Molecular Ophthalmology, Medical and Molecular Genetics Institue (INGEMM) IdiPaz, CIBERER, Hospital Universitario La Paz, 28046 Madrid, Spain; [email protected] * Correspondence: [email protected] Abstract: Background: Congenital aniridia is a complex ocular disorder, usually associated with severe visual impairment, generally caused by mutations on the PAX6 gene. The clinical phenotype of PAX6 mutations is highly variable, making the genotype–phenotype correlations difficult to establish. Methods: we describe the phenotype of eight patients from seven unrelated families Citation: Nieves-Moreno, M.; Noval, with confirmed mutations in PAX6, and very different clinical manifestations.
    [Show full text]
  • Supplementary Materials
    Supplementary Materials + - NUMB E2F2 PCBP2 CDKN1B MTOR AKT3 HOXA9 HNRNPA1 HNRNPA2B1 HNRNPA2B1 HNRNPK HNRNPA3 PCBP2 AICDA FLT3 SLAMF1 BIC CD34 TAL1 SPI1 GATA1 CD48 PIK3CG RUNX1 PIK3CD SLAMF1 CDKN2B CDKN2A CD34 RUNX1 E2F3 KMT2A RUNX1 T MIXL1 +++ +++ ++++ ++++ +++ 0 0 0 0 hematopoietic potential H1 H1 PB7 PB6 PB6 PB6.1 PB6.1 PB12.1 PB12.1 Figure S1. Unsupervised hierarchical clustering of hPSC-derived EBs according to the mRNA expression of hematopoietic lineage genes (microarray analysis). Hematopoietic-competent cells (H1, PB6.1, PB7) were separated from hematopoietic-deficient ones (PB6, PB12.1). In this experiment, all hPSCs were tested in duplicate, except PB7. Genes under-expressed or over-expressed in blood-deficient hPSCs are indicated in blue and red respectively (related to Table S1). 1 C) Mesoderm B) Endoderm + - KDR HAND1 GATA6 MEF2C DKK1 MSX1 GATA4 WNT3A GATA4 COL2A1 HNF1B ZFPM2 A) Ectoderm GATA4 GATA4 GSC GATA4 T ISL1 NCAM1 FOXH1 NCAM1 MESP1 CER1 WNT3A MIXL1 GATA4 PAX6 CDX2 T PAX6 SOX17 HBB NES GATA6 WT1 SOX1 FN1 ACTC1 ZIC1 FOXA2 MYF5 ZIC1 CXCR4 TBX5 PAX6 NCAM1 TBX20 PAX6 KRT18 DDX4 TUBB3 EPCAM TBX5 SOX2 KRT18 NKX2-5 NES AFP COL1A1 +++ +++ 0 0 0 0 ++++ +++ ++++ +++ +++ ++++ +++ ++++ 0 0 0 0 +++ +++ ++++ +++ ++++ 0 0 0 0 hematopoietic potential H1 H1 H1 H1 H1 H1 PB6 PB6 PB7 PB7 PB6 PB6 PB7 PB6 PB6 PB6.1 PB6.1 PB6.1 PB6.1 PB6.1 PB6.1 PB12.1 PB12.1 PB12.1 PB12.1 PB12.1 PB12.1 Figure S2. Unsupervised hierarchical clustering of hPSC-derived EBs according to the mRNA expression of germ layer differentiation genes (microarray analysis) Selected ectoderm (A), endoderm (B) and mesoderm (C) related genes differentially expressed between hematopoietic-competent (H1, PB6.1, PB7) and -deficient cells (PB6, PB12.1) are shown (related to Table S1).
    [Show full text]
  • Drosophila Pax6 Promotes Development of the Entire Eye-Antennal Disc, Thereby Ensuring Proper Adult Head Formation
    PAPER Drosophila Pax6 promotes development of the entire COLLOQUIUM eye-antennal disc, thereby ensuring proper adult head formation Jinjin Zhua, Sneha Palliyila, Chen Ranb, and Justin P. Kumara,1 aDepartment of Biology, Indiana University, Bloomington, IN 47405; and bDepartment of Biology, Stanford University, Stanford, CA 94305 Edited by Ellen V. Rothenberg, California Institute of Technology, Pasadena, CA, and accepted by Editorial Board Member Neil H. Shubin February 17, 2017 (received for review July 26, 2016) Paired box 6 (Pax6) is considered to be the master control gene for molecular battle among GRNs allows for the subdivision of the eye development in all seeing animals studied so far. In vertebrates, eye-antennal disc to be maintained within a single continuous it is required not only for lens/retina formation but also for the cellular field (13–16). Of the GRNs that are known to operate development of the CNS, olfactory system, and pancreas. Although within the eye-antennal disc, the retinal determination (RD) Pax6 plays important roles in cell differentiation, proliferation, and network, which controls eye development, is the best studied (17). patterning during the development of these systems, the underlying At the core of the RD network lie the Paired box 6 (Pax6) genes mechanism remains poorly understood. In the fruit fly, Drosophila eyeless (ey)andtwin of eyeless (toy), the SIX family member sine melanogaster, Pax6 also functions in a range of tissues, including oculis (so), the transcriptional coactivator eyes absent (eya), and the the eye and brain. In this report, we describe the function of Pax6 in Ski/Sno family member dachshund (dac)(17).
    [Show full text]
  • Transcription Factors Define the Neuroanatomical Organization Of
    ORIGINAL RESEARCH ARTICLE published: 14 May 2013 NEUROANATOMY doi: 10.3389/fnana.2013.00007 Transcription factors define the neuroanatomical organization of the medullary reticular formation Paul A. Gray * Department of Anatomy and Neurobiology, Washington University School of Medicine, St. Louis, MO, USA Edited by: The medullary reticular formation contains large populations of inadequately described, Kathleen S. Rockland, MIT, USA excitatory interneurons that have been implicated in multiple homeostatic behaviors Reviewed by: including breathing, viserosensory processing, vascular tone, and pain. Many hindbrain Joan S. Baizer, University of Buffalo, nuclei show a highly stereotyped pattern of localization across vertebrates suggesting USA Ruth Stornetta, University of a strong underlying genetic organization. Whether this is true for neurons within Virginia, USA the reticular regions of hindbrain is unknown. Hindbrain neurons are derived from *Correspondence: distinct developmental progenitor domains each of which expresses distinct patterns of Paul A. Gray, Department of transcription factors (TFs). These neuronal populations have distinct characteristics such Anatomy and Neurobiology, as transmitter identity, migration, and connectivity suggesting developmentally expressed Washington University School of Medicine, Box 8108, 660 S. Euclid TFs might identify unique subpopulations of neurons within the reticular formation. A Ave., St. Louis, MO 63110, USA. fate-mapping strategy using perinatal expression of reporter genes within Atoh1, Dbx1, e-mail: [email protected] Lmx1b,andPtf1a transgenic mice coupled with immunohistochemistry (IHC) and in situ hybridization (ISH) were used to address the developmental organization of a large subset of reticular formation glutamatergic neurons. All hindbrain lineages have relatively large populations that extend the entire length of the hindbrain. Importantly, the location of neurons within each lineage was highly constrained.
    [Show full text]
  • AP-1 in Cell Proliferation and Survival
    Oncogene (2001) 20, 2390 ± 2400 ã 2001 Nature Publishing Group All rights reserved 0950 ± 9232/01 $15.00 www.nature.com/onc AP-1 in cell proliferation and survival Eitan Shaulian1 and Michael Karin*,1 1Laboratory of Gene Regulation and Signal Transduction, Department of Pharmacology, University of California San Diego, 9500 Gilman Drive, La Jolla, California, CA 92093-0636, USA A plethora of physiological and pathological stimuli extensively discussed previously (Angel and Karin, induce and activate a group of DNA binding proteins 1991; Karin, 1995). that form AP-1 dimers. These proteins include the Jun, The mammalian AP-1 proteins are homodimers and Fos and ATF subgroups of transcription factors. Recent heterodimers composed of basic region-leucine zipper studies using cells and mice de®cient in individual AP-1 (bZIP) proteins that belong to the Jun (c-Jun, JunB proteins have begun to shed light on their physiological and JunD), Fos (c-Fos, FosB, Fra-1 and Fra-2), Jun functions in the control of cell proliferation, neoplastic dimerization partners (JDP1 and JDP2) and the closely transformation and apoptosis. Above all such studies related activating transcription factors (ATF2, LRF1/ have identi®ed some of the target genes that mediate the ATF3 and B-ATF) subfamilies (reviewed by (Angel eects of AP-1 proteins on cell proliferation and death. and Karin, 1991; Aronheim et al., 1997; Karin et al., There is evidence that AP-1 proteins, mostly those that 1997; Liebermann et al., 1998; Wisdom, 1999). In belong to the Jun group, control cell life and death addition, some of the Maf proteins (v-Maf, c-Maf and through their ability to regulate the expression and Nrl) can heterodimerize with c-Jun or c-Fos (Nishiza- function of cell cycle regulators such as Cyclin D1, p53, wa et al., 1989; Swaroop et al., 1992), whereas other p21cip1/waf1, p19ARF and p16.
    [Show full text]