DOCSLIB.ORG

The Induction and Early Development of the Midbrain-Hindbrain in The

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
The Induction and Early Development of the Midbrain-Hindbrain in The Institut für Entwicklungsgenetik GSF Forschungszentrum für Umwelt und Gesundheit TUM Zebrafisch Neurogenetik The induction and early development of the midbrain-hindbrain in the embryonic zebrafish Alexandra Tallafuß Vollständiger Abdruck der von der Fakultät Wissenschaftszentrum Weihenstephan für Ernährung, Landnutzung und Umwelt der Technischen Universität München zur Erlangung des akademischen Grads eines Doktors der Naturwissenschaften genehmigten Dissertation. Vorsitzender: Univ.-Prof. Dr. Erwin Grill Prüfer der Dissertation: 1. Univ.-Prof. Dr. Wolfgang Wurst 2. Univ.-Prof. Dr. Alfons Gierl 3. Univ.-Prof. Dr. Heinrich H.D. Meyer Die Dissertation wurde am 21.02.03 bei der Technischen Universität München eingereicht und durch die Fakultät Wissenschaftszentrum Weihenstephan für Ernährung, Landnutzung und Umwelt am 15.04.03 angenommen. TABLE OF CONTENTS i Table of Contents Abstract......................................................................................................................................iv Zusammenfassung......................................................................................................................v Abbreviations used in the text..................................................................................................vii Chapter I: Introduction...............................................................................................................1 1. Neural plate patterning...................................................................................................2 1.1. Neural plate induction..........................................................................................2 1.2. Common features of early neural plate patterning in vertebrates........................5 1.3. Patterning refinement along AP by local signaling centers.................................7 1.4. Early AP neural plate patterning in zebrafish......................................................8 2. The midbrain-hindbrain domain...................................................................................15 2.1. Position and functional derivatives....................................................................15 2.2. The MHD follows an atypical mode of development that responds to an organizer.............................................................................16 2.3. Molecular bases of MHD development and IsO activity...................................18 2.4. Major unanswered questions of MHD development..........................................21 2.5. Formation of the MHD in zebrafish...................................................................22 3. Neurogenesis and bHLH factors...................................................................................24 3.1. Delimiting neurogenesis sites within the vertebrate neural plate: proneural cluster..................................................................................................24 3.2. The neurogenesis process and lateral inhibition.................................................26 3.3. Main actors of the neurogenesis process: bHLH factors....................................27 Chapter II: Aims of the Thesis..................................................................................................32 Chapter III: Results and Discussion..........................................................................................34 1. Bts1 is the earliest selective regulator of pax2.1 in the MHD...................................35 2. Evolutionary consideration about the vertebrate MHD............................................35 3. Signals from the PCP control the size of the six3-positive neuronal cluster.............35 4. The transgenic line herPAC::egfp identifies her5 regulatory elements and allows to follow the fate of the entire prospective MHD.......................................36 5. Molecular and functional analyses of the bHLH factor her5........................................38 6. Identification of a new hairy/E(spl) gene, called him, and preliminary functional results.................................................................................39 7. To do ... .........................................................................................................................43 References.................................................................................................................................45 TABLE OF CONTENTS ii Appendix 1: The zebrafish buttonhead-like factor Bts1 is and early regulator of pax2.1 expression during mid-hindbrain development............................................56 Appendix 2: Formation of the head-trunk boundary in the animal body plan: an evolutionary perspective..........................................................................57 Appendix 3: Selective control of neuronal cluster size at the di-mesencephalic boundary by long-range signaling from the prechordal plate...............................58 Appendix 4: Tracing of her5 progeny in zebrafish transgenics reveals the dynamics of midbrain-hindbrain neurogenesis and maintenance.............................................59 Appendix 5: Regional inhibition of neurogenesis by the zebrafish bHLH transcription factor Her5 shapes the midbrain-hindbrain neuronal differentiation pattern............................................60 Chapter IV: Materials and Methods..........................................................................................61 1. Abbreviations and symbols used in the chapter Materials and Methods..................61 2. Recipes of often used Buffers and Solutions............................................................63 3. Animal model organism: Zebrafish...........................................................................66 3.1. Description and origin................................................................................66 3.2. Animal keeping and breeding.....................................................................67 3.3. Embryos......................................................................................................67 4. Molecular Techniques...............................................................................................67 4.1. Preparation of nucleic acids........................................................................67 4.1.1. Isolation of genomic DNA from embryos...................................67 4.1.2. Isolation of genomic DNA from tail-fins....................................68 4.1.3. Isolation of total RNA from embryos..........................................68 4.1.4. Preparation of cDNA...................................................................68 4.1.5. Plasmid DNA preparation...........................................................68 4.2. Preparation of digoxigenin- or flourescein- labeled anti-sense probes......68 4.3. Amplification of DNA by polymerase chain reaction (PCR)....................69 4.4. PCR purification.........................................................................................69 4.5. Isolation and purification of DNA fragments using Gel Extraction...........69 4.6. Ligation and Transformation......................................................................69 4.7. Transfer and detection of nucleic acids......................................................70 4.7.1. Southern Blot...............................................................................70 4.7.2. Detection of DNA probes............................................................70 4.7.3. Hybridization and detection.........................................................70 4.7.4. Autoradiography..........................................................................71 4.8. Analysis of sequences.................................................................................71 TABLE OF CONTENTS iii 5. Staining of embryos using in situ hybridization (ISH) and immunocytochemistry (IC).....................................................................................71 5.1. Preparation of embryos for ISH and IC......................................................71 5.2. Whole-mount ISH on zebrafish embryos...................................................71 5.2.1. Single color ISH..........................................................................71 5.2.2. Two-color ISH.............................................................................72 5.3. Immunocytochemistry (IC)........................................................................72 5.3.1. Whole-mount IC..........................................................................73 5.3.2. IC after ISH.................................................................................73 5.3.3. IC on cryostat sections.................................................................73 6. Preparation of flatmounts and sections.....................................................................74 6.1. Flatmounts..................................................................................................74 6.2. Cryostat sections.........................................................................................74 6.3. Plastic sections............................................................................................74 7. Scoring of the embryos.............................................................................................74 8. Additional and more detailed description of methods used in this Ph.D. work........74 Danksagung...............................................................................................................................75
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]
  • Auditory Experience Controls the Maturation of Song Discrimination and Sexual Response in Drosophila Xiaodong Li, Hiroshi Ishimoto, Azusa Kamikouchi*
    RESEARCH ARTICLE Auditory experience controls the maturation of song discrimination and sexual response in Drosophila Xiaodong Li, Hiroshi Ishimoto, Azusa Kamikouchi* Graduate School of Science, Nagoya University, Nagoya, Japan Abstract In birds and higher mammals, auditory experience during development is critical to discriminate sound patterns in adulthood. However, the neural and molecular nature of this acquired ability remains elusive. In fruit flies, acoustic perception has been thought to be innate. Here we report, surprisingly, that auditory experience of a species-specific courtship song in developing Drosophila shapes adult song perception and resultant sexual behavior. Preferences in the song-response behaviors of both males and females were tuned by social acoustic exposure during development. We examined the molecular and cellular determinants of this social acoustic learning and found that GABA signaling acting on the GABAA receptor Rdl in the pC1 neurons, the integration node for courtship stimuli, regulated auditory tuning and sexual behavior. These findings demonstrate that maturation of auditory perception in flies is unexpectedly plastic and is acquired socially, providing a model to investigate how song learning regulates mating preference in insects. DOI: https://doi.org/10.7554/eLife.34348.001 Introduction Vocal learning in infants or juvenile birds relies heavily on the early experience of the adult conspe- cific sounds. In humans, early language input is necessary to form the ability of phonetic distinction *For correspondence: and pattern detection in the phase of auditory learning (Doupe and Kuhl, 1999; Kuhl, 2004). [email protected] Because of the strong parallels between speech acquisition of humans and song learning of song- Competing interests: The birds, and the difficulties to investigate the neural mechanisms of human early auditory memory at authors declare that no cellular resolution, songbirds have been used as a predominant model in studying memory formation competing interests exist.
    [Show full text]
  • A Spiking Neuron Classifier Network with a Deep Architecture Inspired By
    A spiking neuron classifier network with a deep architecture inspired by the olfactory system of the honeybee Chris Hausler¨ ∗†, Martin Paul Nawrot∗† and Michael Schmuker∗† ∗Neuroinformatics & Theoretical Neuroscience, Institute of Biology Freie Universitat¨ Berlin, 14195 Berlin, Germany † Bernstein Center for Computational Neuroscience Berlin, 10115 Berlin, Germany Email: [email protected], fmartin.nawrot, [email protected] Abstract—We decompose the honeybee’s olfactory pathway Mushroom into local circuits that represent successive processing stages and body (MB) calyx resemble a deep learning architecture. Using spiking neuronal network models, we infer the specific functional role of these microcircuits in odor discrimination, and measure their con- 2 tribution to the performance of a spiking implementation of a probabilistic classifier, trained in a supervised manner. The entire Kenyon network is based on a network of spiking neurons, suited for Cells (KC) implementation on neuromorphic hardware. Lateral horn 3 I. INTRODUCTION Projection Honeybees can identify odorant stimuli in a quick and neurons (PN) MB-extrinsic neurons To motor reliable fashion [1], [2]. The neuronal circuits involved in neurons odor discrimination are well described at the structural level. 1 Antennal Primary olfactory receptor neurons (ORNs) project to the lobe (AL) From odorant antennal lobe (AL, circuit 1 in Fig. 1). Each ORN is believed receptor neurons to express only one of about 160 olfactory receptor genes [3]. ORNs expressing the same receptor protein converge in the Fig. 1. Processing hierarchy in the honeybee brain. The stages relevant to AL in the same glomerulus, a spherical compartment where this study are numbered for reference.
    [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]
  • 1P MON. PM Cortex and Subcortical Auditory Nuclei
    dynamic role for the vestibular system in orientation and flight control. Laboratory studies of flight behavior under illuminated and dark conditions in both static and rotating obstacle tests were carried out while administering heavy water ͑D2O͒ to bats to impair their vestibular inputs. Eptesicus carried out complex maneuvers through both fixed arrays of wires and a rotating obstacle array using both vision and echolocation, or when guided by echolocation alone. When treated with D2O in combination with lack of visual cues, bats showed considerable decrements in performance. These data indicate that big brown bats use both vision and echolocation to provide spatial registration for head position information generated by the vestibular system. 2:55 1pAB3. Bat’s auditory system: Corticofugal feedback and plasticity. Nobuo Suga ͑Dept. of Biol., Washington Univ., One Brookings Dr., St. Louis, MO 63130͒ The auditory system of the mustached bat consists of physiologically distinct subdivisions for processing different types of biosonar information. It was found that the corticofugal ͑descending͒ auditory system plays an important role in improving and adjusting auditory signal processing. Repetitive acoustic stimulation, cortical electrical stimulation or auditory fear conditioning evokes plastic changes of the central auditory system. The changes are based upon egocentric selection evoked by focused positive feedback associated with lateral inhibition. Focal electric stimulation of the auditory cortex evokes short-term changes in the auditory 1p MON. PM cortex and subcortical auditory nuclei. An increase in a cortical acetylcholine level during the electric stimulation changes the cortical changes from short-term to long-term. There are two types of plastic changes ͑reorganizations͒: centripetal best frequency shifts for expanded reorganization of a neural frequency map and centrifugal best frequency shifts for compressed reorganization of the map.
    [Show full text]
  • Krogh's Principle
    Introduction to Neuroscience: Behavioral Neuroscience Neuroethology, Comparative Neuroscience, Natural Neuroscience Nachum Ulanovsky Department of Neurobiology, Weizmann Institute of Science 2017-2018, 2nd semester Principles of Neuroethology Neuroethology seeks to understand the mechanisms by which the Neurobiology central nervous system controls the Neuroethology Ethology natural behavior of animals. • Focus on Natural behaviors: Choosing to study a well-defined and reproducible yet natural behavior (either Innate or Learned behavior) • Need to study thoroughly the animal’s behavior, including in the field: Neuroethology starts with a good understanding of Ethology. • If you study the animals in the lab, you need to keep them in conditions as natural as possible, to avoid the occurrence of unnatural behaviors. • Krogh’s principle 1 Krogh’s principle August Krogh Nobel prize 1920 “For such a large number of problems there will be some animal of choice or a few such animals on which it can be most conveniently studied. Many years ago when my teacher, Christian Bohr, was interested in the respiratory mechanism of the lung and devised the method of studying the exchange through each lung separately, he found that a certain kind of tortoise possessed a trachea dividing into the main bronchi high up in the neck, and we used to say as a laboratory joke that this animal had been created expressly for the purposes of respiration physiology. I have no doubt that there is quite a number of animals which are similarly "created" for special physiological
    [Show full text]
  • Lateral Inhibition Effects Demonstrate That the Maturational State of Neurons That Then Inhibit Their Neighbors
    522 Lateral Inhibition effects demonstrate that the maturational state of neurons that then inhibit their neighbors. When a the learner’s brain is crucial for the attainment of stimulus (such as a bar of light or any other stim- a language system. ulus) excites a number of neurons in the network Several questions remain open about the mecha- (in this case neurons 4e, 5e, 6e, and 7e), the effect nisms underlying language learning. One issue is of inhibition is to suppress the neurons just out- whether specific aspects of language acquisition side the edge of the bar (3e and 8e) because those should be attributed to language-specific versus neurons are inhibited but not excited. Further, general-purpose learning mechanisms. Another issue because the neurons just inside the edges of the is whether children’s native language can affect the bar (4e and 7e) are excited by light and only inhib- way they think, and whether language is necessary ited by one neighbor, they are especially active. or helpful for the development of human concepts. This leads to perceptual contrast enhancement at borders. Further research showed that lateral inhi- Anna Papafragou bition also applied to overlapping stimuli, and that its strength fell off with distance between the See also Aphasias; Audition: Cognitive Influences; Context Effects in Perception; Speech Perception; Top- interacting stimuli. Down and Bottom-Up Processing; Word Recognition Haldan Keffer Hartline won the Nobel Prize in 1967 for discovering lateral inhibition and its neu- ral correlates. The first inhibitory circuit in the Further Readings nervous system was found in the horseshoe crab (Limulus polyphemus).
    [Show full text]
  • Sensory Receptors A17 (1)
    SENSORY RECEPTORS A17 (1) Sensory Receptors Last updated: April 20, 2019 Sensory receptors - transducers that convert various forms of energy in environment into action potentials in neurons. sensory receptors may be: a) neurons (distal tip of peripheral axon of sensory neuron) – e.g. in skin receptors. b) specialized cells (that release neurotransmitter and generate action potentials in neurons) – e.g. in complex sense organs (vision, hearing, equilibrium, taste). sensory receptor is often associated with nonneural cells that surround it, forming SENSE ORGAN. to stimulate receptor, stimulus must first pass through intervening tissues (stimulus accession). each receptor is adapted to respond to one particular form of energy at much lower threshold than other receptors respond to this form of energy. adequate (s. appropriate) stimulus - form of energy to which receptor is most sensitive; receptors also can respond to other energy forms, but at much higher thresholds (e.g. adequate stimulus for eye is light; eyeball rubbing will stimulate rods and cones to produce light sensation, but threshold is much higher than in skin pressure receptors). when information about stimulus reaches CNS, it produces: a) reflex response b) conscious sensation c) behavior alteration SENSORY MODALITIES Sensory Modality Receptor Sense Organ CONSCIOUS SENSATIONS Vision Rods & cones Eye Hearing Hair cells Ear (organ of Corti) Smell Olfactory neurons Olfactory mucous membrane Taste Taste receptor cells Taste bud Rotational acceleration Hair cells Ear (semicircular
    [Show full text]
  • The Retina the Retina the Pigment Layer Contains Melanin That Prevents Light Reflection Throughout the Globe of the Eye
    The Retina The retina The pigment layer contains melanin that prevents light reflection throughout the globe of the eye. It is also a store of Vitamin A. Rods and Cones Photoreceptors of the nervous system responsible for transforming light energy into the electrical energy of the nervous system. Bipolar cells Link between photoreceptors and ganglion cells. Horizontal cells Provide inhibition between bipolar cells. This is the mechanism of lateral inhibition which is important to edge detection and contrast enhancement. Amacrine cells Transmit excitatory signals from bipolar to ganglion cells. Thought to be important in signalling changes in light intensity. Neural circuitry About 125 rods and 5 cones converge on each optic nerve fibre. In the central portion of the fovea there are no rods. The ratio of cones to optic nerves in the fovea is one. This increases the visual acuity of the fovea. Rods and Cones Photosensitive substance in rods is called rhodopsin and in cones is called iodopsin. The Photochemistry of Vision Rhodopsin and Iodopsin Cis-retinal +Opsin Light sensitive Light Trans-retinal Opsin Light insensitive This is a reversible reaction Cis-retinal +Opsin Light sensitive Vitamin A In the presence of light, the rods and cones are hyperpolarized mv t Hyperpolarizing potential lasts for upto half a second, leading to the perception of fusion of flickering lights. Receptor potential – rod and cone potential Hyperpolarizing receptor potential caused by rhodopsin decomposition. Hyperpolarizing potential lasts for upto half a second, leading to the fusion of flickering lights. Light and dark adaptation in the visual system The eye is capable of vision in conditions of light that vary greatly in intensity The eye adapts dynamically to lighting conditions Light and Dark Adaptation We are able to see in light intensities that vary greatly.
    [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]
  • Small and Dim Target Detection Via Lateral Inhibition Filtering and Artificial Bee Colony Based Selective Visual Attention
    Small and Dim Target Detection via Lateral Inhibition Filtering and Artificial Bee Colony Based Selective Visual Attention Haibin Duan1,2*, Yimin Deng1,2, Xiaohua Wang2, Chunfang Xu1 1 State Key Laboratory of Virtual Reality Technology and Systems, School of Automation Science and Electrical Engineering, Beihang University (formerly Beijing University of Aeronautics and Astronautics), Beijing, P. R. China, 2 Science and Technology on Aircraft Control Laboratory, Beihang University, Beijing, P. R. China Abstract This paper proposed a novel bionic selective visual attention mechanism to quickly select regions that contain salient objects to reduce calculations. Firstly, lateral inhibition filtering, inspired by the limulus’ ommateum, is applied to filter low- frequency noises. After the filtering operation, we use Artificial Bee Colony (ABC) algorithm based selective visual attention mechanism to obtain the interested object to carry through the following recognition operation. In order to eliminate the camera motion influence, this paper adopted ABC algorithm, a new optimization method inspired by swarm intelligence, to calculate the motion salience map to integrate with conventional visual attention. To prove the feasibility and effectiveness of our method, several experiments were conducted. First the filtering results of lateral inhibition filter were shown to illustrate its noise reducing effect, then we applied the ABC algorithm to obtain the motion features of the image sequence. The ABC algorithm is proved to be more robust and effective through the comparison between ABC algorithm and popular Particle Swarm Optimization (PSO) algorithm. Except for the above results, we also compared the classic visual attention mechanism and our ABC algorithm based visual attention mechanism, and the experimental results of which further verified the effectiveness of our method.
    [Show full text]