DOCSLIB.ORG

Expansion, Folding, and Abnormal Lamination of the Chick Optic Tectum After Intraventricular Injections of FGF2

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Expansion, Folding, and Abnormal Lamination of the Chick Optic Tectum After Intraventricular Injections of FGF2 Expansion, folding, and abnormal lamination of the chick optic tectum after intraventricular injections of FGF2 Luke D. McGowan1, Roula A. Alaama, Amanda C. Freise, Johnny C. Huang, Christine J. Charvet, and Georg F. Striedter Department of Neurobiology and Behavior, University of California, Irvine, CA 92697 Edited by John C. Avise, University of California, Irvine, Irvine, CA, and approved May 3, 2012 (received for review February 21, 2012) Comparative research has shown that evolutionary increases in human cortical evolution and expansion in terms of delayed and brain region volumes often involve delays in neurogenesis. How- prolonged precursor proliferation (15, 16). ever, little is known about the influence of such changes on The present study began as an attempt to phenocopy natural subsequent development. To get at this question, we injected variation in telencephalon size among birds. Specifically, we rea- FGF2—which delays cell cycle exit in mammalian neocortex—into soned that FGF2 injections into ventricles of embryonic chicks the cerebral ventricles of chicks at embryonic day (ED) 4. This ma- should, by analogy to the work in mammals (14), increase telen- nipulation alters the development of the optic tectum dramatically. cephalon volume, effectively creating chickens with a telencepha- By ED7, the tectum of FGF2-treated birds is abnormally thin and has lon as large as that of parrots and songbirds (7, 9). However, our a reduced postmitotic layer, consistent with a delay in neurogenesis. FGF2 injections did not significantly alter telencephalon de- FGF2 treatment also increases tectal volume and ventricular surface velopment. Instead, they increased the size of the optic tectum, area, disturbs tectal lamination, and creates small discontinuities disrupted tectal lamination, and created tectal gyri and sulci. in the pia mater overlying the tectum. On ED12, the tectum is still It is tempting to dismiss these induced alterations as mere pa- larger in FGF2-treated embryos than in controls. However, lateral thologies of no evolutionary significance. However, developmental portions of the FGF2-treated tectum now exhibit volcano-like “monsters” have long been used by evolutionary developmental laminar disturbances that coincide with holes in the pia, and the biologists to identify the kind of variation and generative princi- caudomedial tectum exhibits prominent folds. To explain these ples with which natural selection must work (17). Most of this observations, we propose that the tangential expansion of the ven- work has used naturally occurring teratologies, but carefully tricular surface in FGF2-treated tecta outpaces the expansion of the selected developmental manipulations can likewise be useful. pial surface, creating abnormal mechanical stresses. Two alternative Conrad Waddington, for example, argued that experimental per- means of alleviating these stresses are tectal foliation and the for- turbations of development can be used to infer the shape of the mation of pial holes. The latter probably alter signaling gradients epigenetic landscapes that constrain developmental and evolu- required for normal cell migration and may generate abnormal tionary variation (18, 19), even if the results are deleterious. patterns of cerebrospinal fluid flow; both abnormalities would gen- In line with Waddington’s approach, here we describe several erate disturbances in tectal lamination. Overall, our findings suggest changes in tectal development that are induced by FGF2 and that evolutionary expansion of sheet-like, laminated brain regions provide a model to explain them. Our analysis suggests that ex- requires a concomitant expansion of the pia mater. perimental expansion of the optic tectum is disruptive mainly because it is not accompanied by an equivalent expansion of the brain evolution | evolutionary developmental biology | proliferation | pia mater. This mismatch causes tectal foliation and tears holes meninges | cortical folding in pia, which then disrupt laminar development. The presumably maladaptive nature of these alterations probably explains why volutionary increases in brain region volumes are common (1). naturally occurring species differences in optic tectum size are EFor example, the neocortex is disproportionately enlarged in based, as far as we know, on changes in brain patterning, rather primates relative to other mammals, and the telencephalon is than neurogenesis timing (20). Our findings also suggest that disproportionately enlarged in parrots and songbirds relative to the evolutionary expansion of mammalian neocortex must have other birds (1–4). Recent work in evolutionary developmental required a concordant expansion of the neocortical pia. neurobiology has shown that these evolutionary increases in brain region volumes are often caused by delays in cell cycle exit of Results neuronal precursors (5, 6). Among birds, for example, parrots and We first report on changes in tectal volume, ventricular surface songbirds exhibit delayed telencephalic neurogenesis relative to area, and proliferative-zone fraction at embryonic day (ED) chicken-like birds (7–9). Among mammals, cell cycle exit in the 7, 3 d after the FGF2 injections (on ED4). We then describe how neocortex is similarly delayed in primates, which have a dispro- these alterations manifest on ED12. Finally, we describe portionately enlarged neocortex (5, 10–12). Unfortunately, the downstream effects of delayed cell cycle exit on subsequent developmental processes and adult morphology This paper results from the Arthur M. Sackler Colloquium of the National Academy of remain poorly understood. One way to fill this gap in our knowl- Sciences, “In the Light of Evolution VI: Brain and Behavior,” held January 19–21, 2012, at the Arnold and Mabel Beckman Center of the National Academies of Sciences and Engi- edge is to experimentally recreate the key species differences in neering in Irvine, CA. The complete program and audio files of most presentations are the laboratory by means of carefully selected developmental available on the NAS Web site at www.nasonline.org/evolution_vi. manipulations. A good example of this phenocopy approach was Author contributions: L.D.M. and G.F.S. designed research; L.D.M., R.A.A., A.C.F., J.C.H., the creation of transgenic mice with a constitutively active form and C.J.C. performed research; L.D.M., R.A.A., A.C.F., J.C.H., and C.J.C. analyzed data; and of β-catenin that prolongs proliferation, increases neocortical L.D.M. and G.F.S. wrote the paper. volume, and generates cortical folds (13). In another example, it The authors declare no conflict of interest. has been shown that intraventricular injections of FGF2 in rats This article is a PNAS Direct Submission. delay neocortical cell cycle exit, leading to dramatic increases 1To whom correspondence should be addressed. E-mail: [email protected]. in neocortex volume and neuron number (14). Based in part on This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10. these experiments, it is becoming increasingly common to explain 1073/pnas.1201875109/-/DCSupplemental. 10640–10646 | PNAS | June 26, 2012 | vol. 109 | suppl. 1 www.pnas.org/cgi/doi/10.1073/pnas.1201875109 Downloaded by guest on September 26, 2021 qualitative changes in tectal shape and cytoarchitecture that re- Postproliferative cells in the developing brain’s mantle zone sult from FGF2 injection. are less densely packed than their proliferating precursors in the ventricular zone (Fig. 1C). Therefore, the FGF2-induced tectal FGF2 Expands the Tectal Progenitor Pool. On ED7, FGF2-treated thinning is consistent with FGF2 delaying tectal cell cycle exit. embryos exhibit an expanded tectum compared with controls To test this hypothesis, we computed the fraction of all tectal (Fig. 1). Because of substantial variability in absolute volumes at cells that is proliferative, rather than postproliferative. We have this age, we express tectum volume relative to the rest of the previously used this proliferative zone fraction (PZF) measure to brain (minus telencephalon and medulla), as estimated stereo- demonstrate species differences in neurogenesis timing (7, 9). logically (Materials and Methods). By using these methods, we Here we extend the approach by staining ED7 brains with pro- observed a 32% increase in normalized tectum volume [t(11) = liferating cell nuclear antigen (PCNA), a relatively specific 3.9; P < 0.01; n = 13] for the FGF2-treated chicks relative to marker for proliferating cells (21) (Fig. 1 C and D). As very few controls. In contrast, telencephalon volume does not differ PCNA-negative cells were observed within the PCNA-positive significantly between FGF2-treated and control embryos, relative zone (and vice versa), we calculated the tectum’s PZF as the to diencephalon-tegmentum volume [t(11) = 1.3; P = 0.23; n = volume of the PCNA-positive zone divided by total tectum 13]. FGF2 injections also expand the tectum’s ventricular surface volume. Our analysis revealed that the tectum’s PZF is 67% area by 79% [t(11) = 4.9; P < 0.01; n = 13] and reduce tectal larger in FGF2-treated embryos than in controls [t(9) = 7.2; P < thickness by 35% [t(11) = −4.5; P < 0.01; n = 13]. 0.01; n = 11; Fig. 1E). Fig. 1. On ED7, the optic tectum is expanded in FGF2-treated embryos relative to controls, as illustrated here with Giemsa-stained horizontal sections (A and B; anterior is to the left). Staining with antibodies against PCNA (C and D) reveals that FGF2 treatment increases the proportion of proliferating cells in the tectum (i.e., PZF; E). Normalized tectum volume and tectal ventricular surface area are also larger in FGF2-treated birds than in controls, but tectal radial thickness is reduced. SE bars are shown. (Scale bars: A and B, 1 mm; C and D,100μm.) McGowan et al. PNAS | June 26, 2012 | vol. 109 | suppl. 1 | 10641 Downloaded by guest on September 26, 2021 Fig. 2. FGF2-treated embryos on ED12 have a significantly enlarged and abnormal tectum (A and B). Absolute and normalized tectum volumes, as well as the tectum’s ventricular surface, are increased significantly in FGF2-treated chickens relative to controls (C).
Load more

Recommended publications
  • Temporary Disruption of the Retinal Basal Lamina and Its Effect on Retinal Histogenesis
    Developmental Biology 238, 79–96 (2001) doi:10.1006/dbio.2001.0396, available online at http://www.idealibrary.com on Temporary Disruption of the Retinal Basal Lamina and Its Effect on Retinal Histogenesis Willi Halfter,*,1 Sucai Dong,* Manimalha Balasubramani,* and Mark E. Bier† *Department of Neurobiology, University of Pittsburgh, 1402 E Biological Science Tower, Pittsburgh, Pennsylvania 15261; and †Department of Chemistry, Mellon Institute, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, Pennsylvania 15213-2683 An experimental paradigm was devised to remove the retinal basal lamina for defined periods of development: the basal lamina was dissolved by injecting collagenase into the vitreous of embryonic chick eyes, and its regeneration was induced by a chase with mouse laminin-1 and ␣2-macroglobulin. The laminin-1 was essential in reconstituting a new basal lamina and could not be replaced by laminin-2 or collagen IV, whereas the macroglobulin served as a collagenase inhibitor that did not directly contribute to basal lamina regeneration. The regeneration occurred within 6 h after the laminin-1 chase by forming a morphologically complete basal lamina that included all known basal lamina proteins from chick embryos, such as laminin-1, nidogen-1, collagens IV and XVIII, perlecan, and agrin. The temporary absence of the basal lamina had dramatic effects on retinal histogenesis, such as an irreversible retraction of the endfeet of the neuroepithelial cells from the vitreal surface of the retina, the formation of a disorganized ganglion cell layer with an increase in ganglion cells by 30%, and the appearance of multiple retinal ectopias. Finally, basal lamina regeneration was associated with aberrant axons failing to correctly enter the optic nerve.
    [Show full text]
  • In Normal Conditions After Heat Stress
    1 Transcriptome analysis of yamame (Oncorhynchus masou) in normal conditions after heat stress Waraporn Kraitavin1, Kazutoshi Yoshitake1, Yoji Igarashi1, Susumu Mitsuyama1, Shigeharu Kinoshita1, Daisuke Kambayashi2, Shugo Watabe3 and Shuichi Asakawa1* 1 Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo, Tokyo 113-8657, Japan; [email protected] (S.A.) 2 Kobayashi Branch, Miyazaki Prefectural Fisheries Research Institute, Kobayashi, Miyazaki 886-0005, Japan 3 School of Marine Biosciences, Kitasato University, Minami, Sagamihara, Kanagawa 252-0313, Japan; [email protected] * Correspondence: [email protected] (S.A.) 2 A B *HT represents high-temperature tolerant, NT represents non-high-temperature tolerant Figure S1. The pheatmap repeatability analysis of mRNA libraries between samples using the Pearson correlation, (A) gill and (B) fin 3 A B *HT represents high-temperature tolerant, NT represents non-high-temperature tolerant Figure S2. The PCA analysis of mRNA libraries between samples, (A) gill and (B) fin. 4 Table S1. List of the differentially expressed genes of the gill in yamame Gene_id Annotation HT_TPM NT_TPM log2(FoldChange) P - value TRINITY_DN100000_c0_g1_i2 VDAC2 27.57 17.15 0.71 0.00 TRINITY_DN100006_c4_g1_i1 0.65 2.69 -1.93 0.00 TRINITY_DN100021_c0_g1_i2 CXL14 0.13 4.06 -4.96 0.00 TRINITY_DN100027_c4_g2_i1 PEAK1 1.31 4.44 -1.77 0.00 TRINITY_DN100027_c4_g3_i1 0.17 2.38 -3.18 0.00 TRINITY_DN100035_c6_g6_i2 2.13 13.97 -2.68 0.00 TRINITY_DN100055_c1_g2_i1 3BHS 0.22
    [Show full text]
  • Supplementary Table 1: Adhesion Genes Data Set
    Supplementary Table 1: Adhesion genes data set PROBE Entrez Gene ID Celera Gene ID Gene_Symbol Gene_Name 160832 1 hCG201364.3 A1BG alpha-1-B glycoprotein 223658 1 hCG201364.3 A1BG alpha-1-B glycoprotein 212988 102 hCG40040.3 ADAM10 ADAM metallopeptidase domain 10 133411 4185 hCG28232.2 ADAM11 ADAM metallopeptidase domain 11 110695 8038 hCG40937.4 ADAM12 ADAM metallopeptidase domain 12 (meltrin alpha) 195222 8038 hCG40937.4 ADAM12 ADAM metallopeptidase domain 12 (meltrin alpha) 165344 8751 hCG20021.3 ADAM15 ADAM metallopeptidase domain 15 (metargidin) 189065 6868 null ADAM17 ADAM metallopeptidase domain 17 (tumor necrosis factor, alpha, converting enzyme) 108119 8728 hCG15398.4 ADAM19 ADAM metallopeptidase domain 19 (meltrin beta) 117763 8748 hCG20675.3 ADAM20 ADAM metallopeptidase domain 20 126448 8747 hCG1785634.2 ADAM21 ADAM metallopeptidase domain 21 208981 8747 hCG1785634.2|hCG2042897 ADAM21 ADAM metallopeptidase domain 21 180903 53616 hCG17212.4 ADAM22 ADAM metallopeptidase domain 22 177272 8745 hCG1811623.1 ADAM23 ADAM metallopeptidase domain 23 102384 10863 hCG1818505.1 ADAM28 ADAM metallopeptidase domain 28 119968 11086 hCG1786734.2 ADAM29 ADAM metallopeptidase domain 29 205542 11085 hCG1997196.1 ADAM30 ADAM metallopeptidase domain 30 148417 80332 hCG39255.4 ADAM33 ADAM metallopeptidase domain 33 140492 8756 hCG1789002.2 ADAM7 ADAM metallopeptidase domain 7 122603 101 hCG1816947.1 ADAM8 ADAM metallopeptidase domain 8 183965 8754 hCG1996391 ADAM9 ADAM metallopeptidase domain 9 (meltrin gamma) 129974 27299 hCG15447.3 ADAMDEC1 ADAM-like,
    [Show full text]
  • 140503 IPF Signatures Supplement Withfigs Thorax
    Supplementary material for Heterogeneous gene expression signatures correspond to distinct lung pathologies and biomarkers of disease severity in idiopathic pulmonary fibrosis Daryle J. DePianto1*, Sanjay Chandriani1⌘*, Alexander R. Abbas1, Guiquan Jia1, Elsa N. N’Diaye1, Patrick Caplazi1, Steven E. Kauder1, Sabyasachi Biswas1, Satyajit K. Karnik1#, Connie Ha1, Zora Modrusan1, Michael A. Matthay2, Jasleen Kukreja3, Harold R. Collard2, Jackson G. Egen1, Paul J. Wolters2§, and Joseph R. Arron1§ 1Genentech Research and Early Development, South San Francisco, CA 2Department of Medicine, University of California, San Francisco, CA 3Department of Surgery, University of California, San Francisco, CA ⌘Current address: Novartis Institutes for Biomedical Research, Emeryville, CA. #Current address: Gilead Sciences, Foster City, CA. *DJD and SC contributed equally to this manuscript §PJW and JRA co-directed this project Address correspondence to Paul J. Wolters, MD University of California, San Francisco Department of Medicine Box 0111 San Francisco, CA 94143-0111 [email protected] or Joseph R. Arron, MD, PhD Genentech, Inc. MS 231C 1 DNA Way South San Francisco, CA 94080 [email protected] 1 METHODS Human lung tissue samples Tissues were obtained at UCSF from clinical samples from IPF patients at the time of biopsy or lung transplantation. All patients were seen at UCSF and the diagnosis of IPF was established through multidisciplinary review of clinical, radiological, and pathological data according to criteria established by the consensus classification of the American Thoracic Society (ATS) and European Respiratory Society (ERS), Japanese Respiratory Society (JRS), and the Latin American Thoracic Association (ALAT) (ref. 5 in main text). Non-diseased normal lung tissues were procured from lungs not used by the Northern California Transplant Donor Network.
    [Show full text]
  • Supporting Information for Proteomics DOI 10.1002/Pmic.200400942
    Supporting Information for Proteomics DOI 10.1002/pmic.200400942 Wei-Jun Qian, Jon M. Jacobs, David G. Camp II, Matthew E. Monroe, Ronald J. Moore, Marina A. Gritsenko, Steve E. Calvano, Stephen F. Lowry, Wenzhong Xiao, Lyle L. Moldawer, Ronald W. Davis, Ronald G. Tompkins and Richard D. Smith Comparative proteome analyses of human plasma following in vivo lipopolysaccharide administration using multidimensional separations coupled with tandem mass spectrometry ã 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim www.proteomics-journal.de Supplemental Table 1: List of 804 identified plasma proteins. Note: Reference IDs correspond to either SwissProt, NCBI, or PIR database entries. For each protein, one representive peptide sequence and the peptide charge_state, SEQUEST Xcorr, and Delcn are listed. The number of different peptides identifying each specific protein is also indicated. To facilitate comparison of protein abundances between the untreated and treated samples, the numbers of peptide hit, the protein abundance ratio (treated/untreated) calculated from peptide peak area ratios, and the standard deviation of peptide peak area ratios for each protein are also listed. No abundance ratio is shown if the protein has no common peptide identified in the two samples. Whether the same protein ID was reported in reference 16 and reference 27 (195 proteins observed from two different sources) is also indicated. Reference Description # of different Representitive peptide Charge_state Xcorr DelCn peptides Peptide Hits Peptide Protein Standard
    [Show full text]
  • Supplementary Table 2
    Ayrault et al. Supplementary Table S2 Term Count % PValue GO:0007155~cell adhesion 55 13.06% 3.97E-15 1417231_at Cldn2 11.31 CLAUDIN 2 1418153_at Lama1 8.17 LAMININ, ALPHA 1 1434601_at Amigo2 7.78 ADHESION MOLECULE WITH IG LIKE DOMAIN 2 1456214_at Pcdh7 7.26 PROTOCADHERIN 7 1437442_at Pcdh7 6.41 PROTOCADHERIN 7 1445256_at Vcl 6.36 VINCULIN 1427009_at Lama5 5.7 LAMININ, ALPHA 5 1453070_at Pcdh17 5.17 PROTOCADHERIN 17 1428571_at Col9a1 4.59 PROCOLLAGEN, TYPE IX, ALPHA 1 1416039_x_at Cyr61 4.56 CYSTEINE RICH PROTEIN 61 1437932_a_at Cldn1 4.53 CLAUDIN 1 1456397_at Cdh4 4.44 CADHERIN 4 1427010_s_at Lama5 4.23 LAMININ, ALPHA 5 1449422_at Cdh4 3.71 CADHERIN 4 1452784_at Itgav 3.01 INTEGRIN ALPHA V 1435603_at Sned1 2.87 SUSHI, NIDOGEN AND EGF-LIKE DOMAINS 1 1438928_x_at Ninj1 2.85 NINJURIN 1 1441498_at Ptprd 2.69 PROTEIN TYROSINE PHOSPHATASE, RECEPTOR TYPE, D 1419632_at Tecta -33.82 TECTORIN ALPHA 1455056_at Lmo7 -14.83 LIM DOMAIN ONLY 7 1450637_a_at Aebp1 -10.7 AE BINDING PROTEIN 1 1421811_at Thbs1 -10.27 THROMBOSPONDIN 1 1434667_at Col8a2 -8.51 PROCOLLAGEN, TYPE VIII, ALPHA 2 1418304_at Pcdh21 -7.26 PROTOCADHERIN 21 1451454_at Pcdh20 -6.63 PROTOCADHERIN 20 1424701_at Pcdh20 -6.54 PROTOCADHERIN 20 1451758_at Lamc3 -6.15 LAMININ GAMMA 3 1422514_at Aebp1 -5.74 AE BINDING PROTEIN 1 1422694_at Ttyh1 -5.62 TWEETY HOMOLOG 1 (DROSOPHILA) 1425594_at Lamc3 -5.43 LAMININ GAMMA 3 1431152_at Hapln3 -5.39 HYALURONAN AND PROTEOGLYCAN LINK PROTEIN 3 1418424_at Tnfaip6 -5.17 TUMOR NECROSIS FACTOR ALPHA INDUCED PROTEIN 6 1451769_s_at Pcdha4;pcdha10;pcdha1;pcdha2;pcdhac1;pcdha6;pcdha8;pcdhac2;pcdha5;pcdha7;pcdha9;pcdha12;pcdha3-4.99
    [Show full text]
  • Ultrarare Heterozygous Pathogenic Variants of Genes Causing Dominant Forms of Early-Onset Deafness Underlie Severe Presbycusis
    Ultrarare heterozygous pathogenic variants of genes causing dominant forms of early-onset deafness underlie severe presbycusis Sophie Bouchera,b,c,d, Fabienne Wong Jun Taia, Sedigheh Delmaghania, Andrea Lellia, Amrit Singh-Estivaleta,b, Typhaine Duponta, Magali Niasme-Grarea,e, Vincent Michela, Nicolas Wolfff, Amel Bahloula, Yosra Bouyacouba, Didier Bouccarag, Bernard Fraysseh, Olivier Deguineh, Lionel Colleti, Hung Thai-Vana,j,k, Eugen Ionescuj, Jean-Louis Kemenyl, Fabrice Giraudetm,n, Jean-Pierre Lavieilleo, Arnaud Devèzeo, Anne-Laure Roudevitch-Pujolp, Christophe Vincentq, Christian Renardr, Valérie Franco-Vidals, Claire Thibult-Apts, Vincent Darrouzets, Eric Bizaguett, Arnaud Coezt,u,v, Hugues Aschardw, Nicolas Michalskia, Gaëlle M. Lefevrea, Anne Auboisp, Paul Avana,m,n,x,1, Crystel Bonneta,b,1, and Christine Petita,y,1,2 aInstitut de l’Audition, Institut Pasteur, INSERM, 75012 Paris, France; bComplexité du Vivant, Sorbonne Universités, Université Pierre et Marie Curie, Université Paris 06, 75005 Paris, France; cCentre Hospitalier Universitaire, Service d’Oto-Rhino-Laryngologie et Chirurgie Cervico-Faciale, 49100 Angers, France; dFaculté de Médecine, Unité de Formation et de Recherche Santé, Université d’Angers, 49100 Angers, France; eService de Biochimie et Biologie Moléculaire, Hôpital d’Enfants Armand-Trousseau, Assistance Publique-Hôpitaux de Paris (AP-HP), 75012 Paris, France; fUnité Récepteurs-Canaux, Institut Pasteur, 75015 Paris, France; gHôpital Beaujon, Hôpitaux Universitaires Paris Nord val-de-Seine, AP-HP, 92110 Clichy,
    [Show full text]
  • Supplementary Information Mapping the Molecular and Structural
    Tsutsui et al. 1 Supplementary information 2 3 Mapping the molecular and structural specialization of the skin basement 4 membrane for inter-tissue interactions 5 6 Ko Tsutsui, Hiroki Machida, Ritsuko Morita, Asako Nakagawa, Kiyotoshi Sekiguchi, 7 Jeffrey H. Miner, Hironobu Fujiwara* 8 9 *Correspondence: [email protected] 10 1 Tsutsui et al. 11 Supplementary Figure a Lin-, 6 integrin- h Lef1-eGFP/CD34/DAPI CD34 98.2 65.9 Lin-PE-Cy7 Basal cells CD34-eFluor660 DAPI 6 integrin-PE b Lin-, Sca-1-, CD34-, 6 integrin+, Lgr6-eGFP+ 18.0 23.4 51.9 Lin-PE-Cy7 6 integrin-PE Sca1-PerCP-Cy5.5 Lower isthmus DAPI CD34-eFluor660 Lgr6-eGFP 6 integrin/ i Lef1-eGFP/ DAPI 6 integrin Lin-, Sca-1-, CD34-, 6 integrin+, Gli1-eGFP+ c 7.0 25.6 53.9 Lin-PE-Cy7 6 integrin-PE Upper bulge Sca1-PerCP-Cy5.5 DAPI CD34-eFluor660 Gli1-eGFP Lin-, CD34+, 6 integrin+ j d CD34/Pdgfra-H2BeGFP/ 8 integrin/DAPI Pdgfra-H2BeGFP/DAPI 8 integrin/DAPI 65.9 4.2 Lin-PE-Cy7 Mid-bulge CD34-eFluor660 DAPI 6 integrin-PE Lin-, Sca-1-, CD34-, 6 integrin+, Cdh3-eGFPmiddle e 24.9 44.0 35.3 Lin-PE-Cy7 6 integrin-PE Hair germ Sca1-PerCP-Cy5.5 DAPI CD34-eFluor660 Cdh3-eGFP k - - Middle + f Lin , CD34 , 6 integrin , Lef1-eGFP CD34 58.8 Lin-PE-Cy7 22.7 91.4 P2 CD34-eFluor660 CD34-eFluor660 Dermal papilla P1 Dermal papilla DAPI Lef1-eGFP 6 integrin-PE Lef1-eGFP g Lin-, Pdgfra-eGFP+ 56.3 l 0.12 0.20 Pdgfra 1.5 Itga8 GFP 0.15 0.10 1.0 49.6 0.10 0.08 Lin-PE-Cy7 0.5 0.05 0.06 CD34-eFluor660 0.00 0.0 0.04 Pdgfra-eGFP Relative expression (GAPDH = 1) DAPI P1 P2 P1 P2 P1 P2 Pan-dermal fibroblasts m n 30 Pan-dermal Mid-bulge fibroblasts 20 Basal Lower Upperisthmus bulgeMid-bulgeHair germDermalPan-dermal papilla fibroblasts1 Basal 10 0.8 Upper bulge Lower isthmus 0.6 Basal 0.4 0 Upper bulge PC2 (9.5%) 0.2 Hair germ Mid-bulge 0 Lower Dermal -10 isthmus papilla Hair germ -0.2 -0.4 Dermal papilla -0.6 -20 Pan-dermal -0.8 -40 -20 0 20 40 60 fibroblast -1 PC1 (71.3%) Supplementary Fig.
    [Show full text]
  • I. the Retinofugal Pathway of Chick
    CHONDROITIN SULFATE AND KERATAN SULFATE PROTEOGLYCANS IN RETINAL AXON GROWTH AND GUIDANCE A THESIS SUBMITTED TO THE FACULTY OF THE GRADUATE SCHOOL OF THE UNIVERSITY OF MINNESOTA BY Brian Douglas McAdams IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY Steven C. McLoon, Advisor August 2018 © 2018 Brian Douglas McAdams Acknowledgements This was only possible due to the generous support and encouragement of my mentor, Dr. William Kennedy. He led the charge to make sure that this thesis would be completed. My new (and old) committee members have also been very encouraging, and I am deeply grateful. I would also like to thank Jerry Sedgwick for his assistance with his confocal microscopes in BIPL and for producing hypersensitized film for our lab. Also, I must thank the late Dr. Daniel Nordquist for his instruction in multiple photographic and laboratory techniques. I miss his keen insights and thoughtful suggestions (as well as the exasperated eye rolls). Thanks also to David Waid, Al Ernst and all the other McLoon lab members that made my time in that lab memorable. Thanks also to David Redish, Virginia Seybold, Virginia Goettl, Tim Gomez, Cheryl Stucky from Neuroscience, and Gwen, Mona, Ioanna, Rose, Patrick, Steve, Kenji, Shawn and Adam from the Kennedy lab for their encouragement and support. Thank you also to CC, JM, KS, BB and JT for your critical input into the process that made this possible. Thank you also to Willi Halfter for his support and encouragement and for the offer of the post-doc opportunity. Tracey, Lillie and my parents also were endlessly encouraging, and tirelessly supportive throughout my time in graduate school and in the years when I was working before the final push.
    [Show full text]
  • Perkinelmer Genomics to Request the Saliva Swab Collection Kit for Patients That Cannot Provide a Blood Sample As Whole Blood Is the Preferred Sample
    Autism and Intellectual Disability TRIO Panel Test Code TR002 Test Summary This test analyzes 2429 genes that have been associated with Autism and Intellectual Disability and/or disorders associated with Autism and Intellectual Disability with the analysis being performed as a TRIO Turn-Around-Time (TAT)* 3 - 5 weeks Acceptable Sample Types Whole Blood (EDTA) (Preferred sample type) DNA, Isolated Dried Blood Spots Saliva Acceptable Billing Types Self (patient) Payment Institutional Billing Commercial Insurance Indications for Testing Comprehensive test for patients with intellectual disability or global developmental delays (Moeschler et al 2014 PMID: 25157020). Comprehensive test for individuals with multiple congenital anomalies (Miller et al. 2010 PMID 20466091). Patients with autism/autism spectrum disorders (ASDs). Suspected autosomal recessive condition due to close familial relations Previously negative karyotyping and/or chromosomal microarray results. Test Description This panel analyzes 2429 genes that have been associated with Autism and ID and/or disorders associated with Autism and ID. Both sequencing and deletion/duplication (CNV) analysis will be performed on the coding regions of all genes included (unless otherwise marked). All analysis is performed utilizing Next Generation Sequencing (NGS) technology. CNV analysis is designed to detect the majority of deletions and duplications of three exons or greater in size. Smaller CNV events may also be detected and reported, but additional follow-up testing is recommended if a smaller CNV is suspected. All variants are classified according to ACMG guidelines. Condition Description Autism Spectrum Disorder (ASD) refers to a group of developmental disabilities that are typically associated with challenges of varying severity in the areas of social interaction, communication, and repetitive/restricted behaviors.
    [Show full text]
  • A Novel Mutation in the TECTA Gene in a Chinese Family with Autosomal Dominant Nonsyndromic Hearing Loss
    A Novel Mutation in the TECTA Gene in a Chinese Family with Autosomal Dominant Nonsyndromic Hearing Loss Yu Su1,2., Wen-Xue Tang3., Xue Gao1,4., Fei Yu1, Zhi-Yao Dai5, Jian-Dong Zhao1,YuLu1, Fei Ji1, Sha-Sha Huang1, Yong-Yi Yuan1, Ming-Yu Han1,2, Yue-Shuai Song1,2, Yu-Hua Zhu1, Dong-Yang Kang1, Dong-Yi HAN1*, Pu Dai1,2* 1 Department of Otorhinolaryngology, Head and Neck Surgery, PLA General Hospital, Beijing, P. R. China, 2 Department of Otorhinolaryngology, Hainan Branch of PLA General Hospital, Sanya, P. R. China, 3 Department of Otolaryngology, Emory University School of Medicine, Atlanta, Georgia, 4 Department of Otorhinolaryngology, the Second Artillery General Hospital, Beijing, P. R. China, 5 Department of Otorhinolaryngology, the First Affiliated Hospital of PLA General Hospital, Beijing, P. R. China Abstract TECTA-related deafness can be inherited as autosomal-dominant nonsyndromic deafness (designated DFNA) or as the autosomal-recessive version. The a-tectorin protein, which is encoded by the TECTA gene, is one of the major components of the tectorial membrane in the inner ear. Using targeted DNA capture and massively parallel sequencing (MPS), we screened 42 genes known to be responsible for human deafness in a Chinese family (Family 3187) in which common deafness mutations had been ruled out as the cause, and identified a novel mutation, c.257–262CCTTTC.GCT (p. Ser86Cys; p. Pro88del) in exon 3 of the TECTA gene in the proband and his extended family. All affected individuals in this family had moderate down-sloping hearing loss across all frequencies. To our knowledge, this is the second TECTA mutation identified in Chinese population.
    [Show full text]
  • Overview of the Matrisome—An Inventory of Extracellular Matrix Constituents and Functions
    Downloaded from http://cshperspectives.cshlp.org/ on September 24, 2021 - Published by Cold Spring Harbor Laboratory Press Overview of the Matrisome—An Inventory of Extracellular Matrix Constituents and Functions Richard O. Hynes and Alexandra Naba Howard Hughes Medical Institute, Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139 Correspondence: [email protected] Completion of genome sequences for many organisms allows a reasonably complete definition of the complement of extracellular matrix (ECM) proteins. In mammals this “core matrisome” comprises 300 proteins. In addition there are large numbers of ECM- modifying enzymes, ECM-binding growth factors, and other ECM-associated proteins. These different categories of ECM and ECM-associated proteins cooperate to assemble and remodel extracellular matrices and bind to cells through ECM receptors. Togetherwith recep- tors for ECM-bound growth factors, they provide multiple inputs into cells to control survival, proliferation, differentiation, shape, polarity, and motility of cells. The evolution of ECM pro- teins was key in the transition to multicellularity, the arrangement of cells into tissue layers, and the elaboration of novel structures during vertebrate evolution. This key role of ECM is reflected in the diversity of ECM proteins and the modular domain structures of ECM proteins both allow their multiple interactions and, during evolution, development of novel protein architectures. he term extracellular matrix (ECM) means make up basement membranes. Biochemistry Tsomewhat different things to different of native ECM was, and still is, impeded by people (Hay 1981, 1991; Mecham 2011). Light the fact that the ECM is, by its very nature, and electron microscopy show that extracellular insoluble and is frequently cross-linked.
    [Show full text]