DOCSLIB.ORG

Dysregulation of Rho Gtpases in the Apix/Arhgef6 Mouse Model of X

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Dysregulation of Rho Gtpases in the Apix/Arhgef6 Mouse Model of X Zurich Open Repository and Archive University of Zurich Main Library Strickhofstrasse 39 CH-8057 Zurich www.zora.uzh.ch Year: 2011 Dysregulation of Rho GTPases in the ฀Pix/Arhgef6 mouse model of X-linked intellectual disability is paralleled by impaired structural and synaptic plasticity and cognitive deficits Ramakers, G J A ; Wolfer, D ; Rosenberger, G ; Kuchenbecker, K ; Kreienkamp, H J ; Prange-Kiel, J ; Rune, G ; Richter, K ; Langnaese, K ; Masneuf, S ; Bösl, M R ; Fischer, K D ; Krugers, H J ; Lipp, H P ; van Galen, E ; Kutsche, K Abstract: Mutations in the ARHGEF6 gene, encoding the guanine nucleotide exchange factor ฀PIX/Cool- 2 for the Rho GTPases Rac1 and Cdc42, cause X-linked intellectual disability (ID) in humans. We show here that ฀Pix/Arhgef6 is primarily expressed in neuropil regions of the hippocampus. To study the role of ฀Pix/Arhgef6 in neuronal development and plasticity and gain insight into the pathogenic mechanisms underlying ID, we generated ฀Pix/Arhgef6-deficient mice. Gross brain structure in these mice appeared to be normal; however, analysis of Golgi-Cox-stained pyramidal neurons revealed an increase in both dendritic length and spine density in the hippocampus, accompanied by an overall loss in spine synapses. Early-phase long-term potentiation was reduced and long-term depression was increased in the CA1 hippocampal area of ฀Pix/Arhgef6-deficient animals. Knockout animals exhibited impaired spatial and complex learning and less behavioral control in mildly stressful situations, suggesting that this model mimics the human ID phenotype. The structural and electrophysiological alterations in the hippocampus were accompanied by a significant reduction in active Rac1 and Cdc42, but not RhoA. In conclusion, we suggest that imbalance in activity of different Rho GTPases may underlie altered neuronal connectivity and impaired synaptic function and cognition in ฀Pix/Arhgef6 knockout mice. DOI: https://doi.org/10.1093/hmg/ddr457 Posted at the Zurich Open Repository and Archive, University of Zurich ZORA URL: https://doi.org/10.5167/uzh-56495 Journal Article Published Version Originally published at: Ramakers, G J A; Wolfer, D; Rosenberger, G; Kuchenbecker, K; Kreienkamp, H J; Prange-Kiel, J; Rune, G; Richter, K; Langnaese, K; Masneuf, S; Bösl, M R; Fischer, K D; Krugers, H J; Lipp, H P; van Galen, E; Kutsche, K (2011). Dysregulation of Rho GTPases in the ฀Pix/Arhgef6 mouse model of X-linked intellectual disability is paralleled by impaired structural and synaptic plasticity and cognitive deficits. Human Molecular Genetics, 21(2):268-286. DOI: https://doi.org/10.1093/hmg/ddr457 Human Molecular Genetics, 2012, Vol. 21, No. 2 268–286 doi:10.1093/hmg/ddr457 Advance Access published on October 11, 2011 Dysregulation of Rho GTPases in the aPix/Arhgef6 mouse model of X-linked intellectual disability is paralleled by impaired structural and synaptic plasticity and cognitive deficits Ger J.A. Ramakers1,2,{, David Wolfer3,4,{, Georg Rosenberger5,{, Kerstin Kuchenbecker5, Hans-Ju¨ rgen Kreienkamp5, Janine Prange-Kiel6,{, Gabriele Rune6, Karin Richter7, Kristina Langnaese7, Sophie Masneuf3, Michael R. Bo¨ sl9,}, Klaus-Dieter Fischer7,8, Harm J. Krugers10, Hans-Peter Lipp2, Elly van Galen1 and Kerstin Kutsche5,∗ 1Department of Neurons and Networks, Netherlands Institute for Neuroscience, an Institute of the Royal Netherlands Academy of Arts and Sciences, Amsterdam, The Netherlands, 2Department of Developmental Psychology, Faculty of Behavioral and Social Sciences, University of Amsterdam, Amsterdam, The Netherlands, 3Institute of Anatomy and Zurich Center for Integrative Human Physiology (ZIHP), University of Zurich, Switzerland, 4Institute of Human Movement Sciences and Sport, ETH Zu¨rich, Switzerland, 5Institut fu¨r Humangenetik, Universita¨tsklinikum Hamburg- Eppendorf, Hamburg, Germany, 6Institute of Anatomy I: Cellular Neurobiology, University Medical Center Hamburg- Eppendorf, Hamburg, Germany, 7Institute of Biochemistry and Cell Biology and 8Division of Functional Genomics and Medical Toponomics at the Centre for Cellular Imaging and Innovative Disease Models, Otto von Guericke University Magdeburg, Magdeburg, Germany, 9Center for Molecular Neurobiology, Hamburg, Germany and 10Swammerdam Institute for Life Sciences, Center for Neuroscience, Amsterdam, The Netherlands Received August 26, 2011; Revised and Accepted September 29, 2011 Mutations in the ARHGEF6 gene, encoding the guanine nucleotide exchange factor aPIX/Cool-2 for the Rho GTPases Rac1 and Cdc42, cause X-linked intellectual disability (ID) in humans. We show here that aPix/Arhgef6 is primarily expressed in neuropil regions of the hippocampus. To study the role of aPix/Arhgef6 in neuronal development and plasticity and gain insight into the pathogenic mechanisms underlying ID, we generated aPix/Arhgef6-deficient mice. Gross brain structure in these mice appeared to be normal; however, analysis of Golgi-Cox-stained pyramidal neurons revealed an increase in both dendritic length and spine density in the hippocampus, accompanied by an overall loss in spine synapses. Early-phase long-term potentiation was reduced and long-term depression was increased in the CA1 hippocampal area of aPix/Arhgef6-deficient animals. Knockout animals exhibited impaired spatial and complex learning and less behavioral control in mildly stressful situa- tions, suggesting that this model mimics the human ID phenotype. The structural and electrophysio- logical alterations in the hippocampus were accompanied by a significant reduction in active Rac1 and Cdc42, but not RhoA. In conclusion, we suggest that imbalance in activity of different Rho GTPases may underlie altered neuronal connectivity and impaired synaptic function and cognition in aPix/Arhgef6 knockout mice. ∗To whom correspondence should be addressed at: Institut fu¨r Humangenetik, Universita¨tsklinikum Hamburg-Eppendorf, Campus Forschung, Martinistraße 52, 20246 Hamburg, Germany. Tel: +49 40741054597; Fax: +49 40741055138; Email: [email protected] †The authors wish it to be known that, in their opinion, the first three authors should be regarded as joint First Authors. ‡Present address: Department of Cell Biology, University of Texas, Southwestern Medical Center at Dallas, Texas, USA. }Present address: MPI of Neurobiology, Martinsried, Germany. Published by Oxford University Press 2011. Downloaded from https://academic.oup.com/hmg/article-abstract/21/2/268/661872 by University of Zurich user on 02 August 2018 Human Molecular Genetics, 2012, Vol. 21, No. 2 269 Downloaded from https://academic.oup.com/hmg/article-abstract/21/2/268/661872 by University of Zurich user on 02 August 2018 270 Human Molecular Genetics, 2012, Vol. 21, No. 2 INTRODUCTION demonstrated only in vitro and underlies a complex allosteric regulation (28–34). Rho GTPases belong to the family of small GTPases which act aPix/Arhgef6 transcripts have been detected in rat hippo- as molecular switches by cycling between an active, campal CA1–CA3 cell layers and ectopically GTP-bound and an inactive, GDP-bound state. The activity expressed aPix/Arhgef6 colocalizes with postsynaptic state of Rho GTPases is tightly controlled by positive and nega- density (PSD) -95 at postsynaptic densities of excitatory tive regulators. Guanine nucleotide exchange factors (GEFs) synapses in cultured neurons (35). Knockdown of aPix/ activate Rho proteins by mediating the exchange of GDP for Arhgef6 results in abnormal spine morphology in hippocampal GTP (1), whereas GTPase-activating proteins (GAPs) stimulate organotypic slice cultures. The active form of Pak3 rescues the the low intrinsic GTP hydrolysis rate (2), thereby negatively phenotype induced by aPix/Arhgef6 downregulation, suggest- regulating Rho signaling. In the active state, Rho GTPases ing that aPix/Arhgef6 acts upstream of Pak3 in regulating exert their effect on various cellular functions. The most exten- spine morphogenesis and plasticity of synaptic networks (35). sively studied Rho GTPases Rac1, Cdc42 and RhoA are best To establish the role of the X-linked ID gene aPIX/ known for their role in controlling dynamics of the actin cyto- ARHGEF6 in vivo, we generated and characterized constitu- skeleton (3,4). Because actin filaments make up the structural tional aPix/Arhgef6 knockout mice. As we found aPix/ framework of dendrites, axons and dendritic spines, Rho signal- Arhgef6 to be highly expressed in the hippocampal formation ing pathways have been suggested to be critically important for and the hippocampus has a major role in learning and memory neurite outgrowth and branching, axon pathfinding and dendrit- (36), we studied synaptic morphology and long-term potenti- ic spine morphogenesis (5–9). ation (LTP) as well as hippocampus-dependent cognitive The importance of Rho-dependent signaling in neuronal behavior in aPix/Arhgef6-deficient mice. development and function has been highlighted by the identi- fication of mutations in Rho-linked genes causing intellectual disability (ID) in humans (10–12). ID is the most common RESULTS developmental disability in children and is defined by a a 2/Y global deficiency of cognitive abilities and adaptive skills Pix/Arhgef6 mice are viable and show no changes in (13). It has been proposed that patients with ID show a gross brain morphology deficit in neuronal network connectivity, resulting in abnormal The targeting strategy for aPix/Arhgef6 knockout mice was information processing (14). There is evidence that the cere- designed to abolish expression of aPix/Arhgef6 resulting in a
Load more

Recommended publications
  • Cdc42 and Rac1 Activity Is Reduced in Human Pheochromocytoma and Correlates with FARP1 and ARHGEF1 Expression
    234 P Croisé et al. Rho-GTPase activity in human 23:4 281–293 Research pheochromocytoma Cdc42 and Rac1 activity is reduced in human pheochromocytoma and correlates with FARP1 and ARHGEF1 expression Pauline Croisé1, Sébastien Houy1, Mathieu Gand1, Joël Lanoix2, Valérie Calco1, Petra Tóth1, Laurent Brunaud3, Sandra Lomazzi4, Eustache Paramithiotis2, Daniel Chelsky2, Stéphane Ory1,* and Stéphane Gasman1,* Correspondence 1Institut des Neurosciences Cellulaires et Intégratives (INCI), CNRS UPR 3212, Strasbourg, France should be addressed 2Caprion Proteome, Inc., Montréal, Québec, Canada to S Ory or S Gasman 3Service de Chirurgie Digestive, Hépato-bilaire et Endocrinienne, CHRU Nancy, Hôpitaux de Brabois, Email Vandoeuvre les Nancy, France [email protected] or 4Centre de Ressources Biologiques (CRB), CHRU Nancy, Hôpitaux de Brabois, Vandoeuvres les Nancy, France [email protected] *(S Ory and S Gasman contributed equally to this work) Abstract Among small GTPases from the Rho family, Cdc42, Rac, and Rho are well known to mediate Key Words a large variety of cellular processes linked with cancer biology through their ability to cycle f Rho-GTPases between an inactive (GDP-bound) and an active (GTP-bound) state. Guanine nucleotide f pheochromocytoma exchange factors (GEFs) stimulate the exchange of GDP for GTP to generate the activated f mass spectrometry form, whereas the GTPase-activating proteins (GAPs) catalyze GTP hydrolysis, leading to f Rho-GEF Endocrine-Related Cancer Endocrine-Related the inactivated form. Modulation of Rho GTPase activity following altered expression of Rho-GEFs and/or Rho-GAPs has already been reported in various human tumors. However, nothing is known about the Rho GTPase activity or the expression of their regulators in human pheochromocytomas, a neuroendocrine tumor (NET) arising from chromaffin cells of the adrenal medulla.
    [Show full text]
  • Focus on Cdc42 in Breast Cancer: New Insights, Target Therapy Development and Non-Coding Rnas
    Review Focus on Cdc42 in Breast Cancer: New Insights, Target Therapy Development and Non-Coding RNAs Yu Zhang †, Jun Li †, Xing-Ning Lai, Xue-Qiao Jiao, Jun-Ping Xiong and Li-Xia Xiong * Department of Pathophysiology, Jiangxi Province Key Laboratory of Tumor Pathogenesis and Molecular Pathology, Medical College, Nanchang University, 461 Bayi Road, Nanchang 330006, China; [email protected] (Y.Z.); [email protected] (J.L.); [email protected] (X.-N.L.); [email protected] (X.-Q.J.); [email protected] (J.-P.X.) * Correspondence: [email protected]; Tel.: +86-791-8636-0556 † These authors contributed equally to this work. Received: 30 December 2018; Accepted: 8 February 2019; Published: 11 February 2019 Abstract: Breast cancer is the most common malignant tumors in females. Although the conventional treatment has demonstrated a certain effect, some limitations still exist. The Rho guanosine triphosphatase (GTPase) Cdc42 (Cell division control protein 42 homolog) is often upregulated by some cell surface receptors and oncogenes in breast cancer. Cdc42 switches from inactive guanosine diphosphate (GDP)-bound to active GTP-bound though guanine-nucleotide- exchange factors (GEFs), results in activation of signaling cascades that regulate various cellular processes such as cytoskeletal changes, proliferation and polarity establishment. Targeting Cdc42 also provides a strategy for precise breast cancer therapy. In addition, Cdc42 is a potential target for several types of non-coding RNAs including microRNAs and lncRNAs. These non-coding RNAs is extensively involved in Cdc42-induced tumor processes, while many of them are aberrantly expressed. Here, we focus on the role of Cdc42 in cell morphogenesis, proliferation, motility, angiogenesis and survival, introduce the Cdc42-targeted non-coding RNAs, as well as present current development of effective Cdc42-targeted inhibitors in breast cancer.
    [Show full text]
  • Individual Protomers of a G Protein-Coupled Receptor Dimer Integrate Distinct Functional Modules
    OPEN Citation: Cell Discovery (2015) 1, 15011; doi:10.1038/celldisc.2015.11 © 2015 SIBS, CAS All rights reserved 2056-5968/15 ARTICLE www.nature.com/celldisc Individual protomers of a G protein-coupled receptor dimer integrate distinct functional modules Nathan D Camp1, Kyung-Soon Lee2, Jennifer L Wacker-Mhyre2, Timothy S Kountz2, Ji-Min Park2, Dorathy-Ann Harris2, Marianne Estrada2, Aaron Stewart2, Alejandro Wolf-Yadlin1, Chris Hague2 1Department of Genome Sciences, University of Washington School of Medicine, Seattle, WA, USA; 2Department of Pharmacology, University of Washington School of Medicine, Seattle, WA, USA Recent advances in proteomic technology reveal G-protein-coupled receptors (GPCRs) are organized as large, macromolecular protein complexes in cell membranes, adding a new layer of intricacy to GPCR signaling. We previously reported the α1D-adrenergic receptor (ADRA1D)—a key regulator of cardiovascular, urinary and CNS function—binds the syntrophin family of PDZ domain proteins (SNTA, SNTB1, and SNTB2) through a C-terminal PDZ ligand inter- action, ensuring receptor plasma membrane localization and G-protein coupling. To assess the uniqueness of this novel GPCR complex, 23 human GPCRs containing Type I PDZ ligands were subjected to TAP/MS proteomic analysis. Syntrophins did not interact with any other GPCRs. Unexpectedly, a second PDZ domain protein, scribble (SCRIB), was detected in ADRA1D complexes. Biochemical, proteomic, and dynamic mass redistribution analyses indicate syntrophins and SCRIB compete for the PDZ ligand, simultaneously exist within an ADRA1D multimer, and impart divergent pharmacological properties to the complex. Our results reveal an unprecedented modular dimeric architecture for the ADRA1D in the cell membrane, providing unexpected opportunities for fine-tuning receptor function through novel protein interactions in vivo, and for intervening in signal transduction with small molecules that can stabilize or disrupt unique GPCR:PDZ protein interfaces.
    [Show full text]
  • The Mental Retardation Protein PAK3 Contributes to Synapse Formation and Plasticity in Hippocampus
    10816 • The Journal of Neuroscience, December 1, 2004 • 24(48):10816–10825 Development/Plasticity/Repair The Mental Retardation Protein PAK3 Contributes to Synapse Formation and Plasticity in Hippocampus Bernadett Boda, Stefano Alberi, Irina Nikonenko, Roxanne Node-Langlois, Pascal Jourdain, Marlyse Moosmayer, Lorena Parisi-Jourdain, and Dominique Muller Department of Basic Neuroscience, Centre Medical Universitaire, 1211 Geneva 4, Switzerland Mutations of the gene coding for PAK3 (p21-activated kinase 3) are associated with X-linked, nonsyndromic forms of mental retardation (MRX) in which the only distinctive clinical feature is the cognitive deficit. The mechanisms through which PAK3 mutation produces the mental handicap remain unclear, although an involvement in the mechanisms that regulate the formation or plasticity of synaptic networks has been proposed. Here we show, using a transient transfection approach, that antisense and small interfering RNA-mediated suppression of PAK3 or expression of a dominant-negative PAK3 carrying the human MRX30 mutation in rat hippocampal organotypic slice cultures results in the formation of abnormally elongated dendritic spines and filopodia-like protrusions and a decrease in mature spine synapses. Ultrastructural analysis of the changes induced by expression of PAK3 carrying the MRX30 mutation reveals that many elongated spines fail to express postsynaptic densities or contact presynaptic terminals. These defects are associated with a reduced spontaneous activity, altered expression of AMPA-type glutamate receptors, and defective long-term potentiation. Together, these data identify PAK3 as a key regulator of synapse formation and plasticity in the hippocampus and support interpretations that these defects might contribute to the cognitive deficits underlying this form of mental retardation.
    [Show full text]
  • A De Novo Mutation in the X-Linked PAK3 Gene Is the Underlying Cause of Intellectual Disability and Macrocephaly in Monozygotic Twins
    European Journal of Medical Genetics xxx (2017) 1e5 Contents lists available at ScienceDirect European Journal of Medical Genetics journal homepage: http://www.elsevier.com/locate/ejmg A de novo mutation in the X-linked PAK3 gene is the underlying cause of intellectual disability and macrocephaly in monozygotic twins Jozef Hertecant a, b, Makanko Komara c, Aslam Nagi a, Olfat Al-Zaabi d, Waseem Fathallah e, Hong Cui f, Yaping Yang f, Christine M. Eng f, Mohammad Al Sorkhy g, * Mohammad A. Ghattas g, Lihadh Al-Gazali b, Bassam R. Ali c, a Department of Paediatrics, Tawam Hospital, Al-Ain, United Arab Emirates b Department of Paediatrics, College of Medicine and Health Sciences, United Arab Emirates University, Al-Ain, United Arab Emirates c Department of Pathology, College of Medicine and Health Sciences, United Arab Emirates University, Al-Ain, United Arab Emirates d Fujairah Hospital, Fujairah, United Arab Emirates e Mafraq Hospital, Abu Dhabi, United Arab Emirates f Department of Molecular and Human Genetics, Baylor College of Medicine, Baylor Miraca Genetics Laboratories, Houston, TX 77030, USA g College of Pharmacy, Al Ain University of Science and Technology, Al-Ain, United Arab Emirates article info abstract Article history: Pathogenic variants in theP21 protein (Cdc42/Rac)-activated kinase 3gene (PAK3) lead to a rare non Received 29 May 2016 syndromic X-linked intellectual disability. The protein encoded by this gene forms an activated complex Received in revised form with GTP-bound RAS-like (P21), CDC2 and RAC1 proteins which then mediates a variety of cellular 17 January 2017 processes. So far, mutations in PAK3 gene have been reported in few families affected with intellectual Accepted 18 January 2017 disability associated with neurological manifestations such as speech defect, behavioral problem, brain Available online xxx structural abnormalities, microcephaly and cerebral palsy.
    [Show full text]
  • Whole Exome Sequencing of Thymic Neuroendocrine Tumor With
    176:2 Y Li, Y Peng and others Sequencing thymic 176:2 187–194 Clinical Study neuroendocrine tumor Whole exome sequencing of thymic neuroendocrine tumor with ectopic ACTH syndrome Yanli Li1,*, Ying Peng1,*, Xiuli Jiang1, Yulong Cheng3, Weiwei Zhou1, Tingwei Su1, Jing Xie2, Xu Zhong1, Dalong Song1, Luming Wu1, Liwen Fan1, Min Li1, Jie Hong1, Weiqing Wang1, Guang Ning1,3 and Yanan Cao1 1Shanghai Clinical Center for Endocrine and Metabolic Diseases, Shanghai Key Laboratory for Endocrine Tumors and 2Department of Pathology, Rui-Jin Hospital, Shanghai Jiao-Tong University School of Medicine, Shanghai, China, and 3Laboratory of Endocrinology and Metabolism, Institute of Health Correspondence Sciences, Shanghai Institutes for Biological Sciences (SIBS), Chinese Academy of Sciences (CAS) & should be addressed Shanghai Jiao-Tong University School of Medicine (SJTUSM), Shanghai, China to Y Cao *(Y Li and Y Peng contributed equally to this work) Email [email protected] Abstract Objective: Thymic neuroendocrine tumor is the second-most prevalent cause of ectopic adrenocorticotropic hormone (ACTH) syndrome (EAS), which is a rare disease characterized by ectopic ACTH oversecretion from nonpituitary tumors. However, the genetic abnormalities of thymic neuroendocrine tumors with EAS remain largely unknown. We aim to elucidate the genetic abnormalities and identify the somatic mutations of potential tumor-related genes of thymic neuroendocrine tumors with EAS by whole exome sequencing. Design and methods: Nine patients with thymic neuroendocrine tumors with EAS who were diagnosed at Shanghai Clinical Center for Endocrine and Metabolic Diseases in Ruijin Hospital between 2002 and 2014 were enrolled. We performed whole exome sequencing on the DNA obtained from thymic neuroendocrine tumors and matched peripheral blood using the Hiseq2000 platform.
    [Show full text]
  • Identification and Bioinformatics Analysis of Overlapping Differentially Expressed Genes in Depression, Papillary Thyroid Cancer and Uterine Fibroids
    4810 EXPERIMENTAL AND THERAPEUTIC MEDICINE 15: 4810-4816, 2018 Identification and bioinformatics analysis of overlapping differentially expressed genes in depression, papillary thyroid cancer and uterine fibroids HANXIAO TANG1 and YONGSHENG ZHANG2 1Department of Pharmacy, Affiliated Tongde Hospital of Zhejiang Chinese Medical University, Hangzhou, Zhejiang 310012; 2The Diagnostic Institute of Chinese Medicine, School of Basic Medicine, Zhejiang Chinese Medical University, Hangzhou, Zhejiang 310053, P.R. China Received May 9, 2017; Accepted October 26, 2017 DOI: 10.3892/etm.2018.6023 Abstract. It is hypothesized that there may be common potential genetic interconnections between depression, PTC and characteristics between the genetic regulatory networks of UF, which may be beneficial for understanding their underlying different diseases. To identify these potential similarities, coregulatory mechanisms and contributing to the development analysis of overlapping differentially expressed genes (DEGs) of homeotherapy based on bioinformatics prediction. in several diseases, which are believed to be associated in traditional Chinese medicine (TCM) was performed in the Introduction present study. The gene expression profiles associated with depression, papillary thyroid carcinoma (PTC) and uterine Molecular biology methods, including DNA microarrays, fibroids (UF) were preliminarily analyzed using Gene are being increasingly used for high-throughput gene expres- Expression Omnibus 2R tools. Gene Ontology enrichment sion analysis and to assess
    [Show full text]
  • Gene Networks Activated by Specific Patterns of Action Potentials in Dorsal Root Ganglia Neurons Received: 10 August 2016 Philip R
    www.nature.com/scientificreports OPEN Gene networks activated by specific patterns of action potentials in dorsal root ganglia neurons Received: 10 August 2016 Philip R. Lee1,*, Jonathan E. Cohen1,*, Dumitru A. Iacobas2,3, Sanda Iacobas2 & Accepted: 23 January 2017 R. Douglas Fields1 Published: 03 March 2017 Gene regulatory networks underlie the long-term changes in cell specification, growth of synaptic connections, and adaptation that occur throughout neonatal and postnatal life. Here we show that the transcriptional response in neurons is exquisitely sensitive to the temporal nature of action potential firing patterns. Neurons were electrically stimulated with the same number of action potentials, but with different inter-burst intervals. We found that these subtle alterations in the timing of action potential firing differentially regulates hundreds of genes, across many functional categories, through the activation or repression of distinct transcriptional networks. Our results demonstrate that the transcriptional response in neurons to environmental stimuli, coded in the pattern of action potential firing, can be very sensitive to the temporal nature of action potential delivery rather than the intensity of stimulation or the total number of action potentials delivered. These data identify temporal kinetics of action potential firing as critical components regulating intracellular signalling pathways and gene expression in neurons to extracellular cues during early development and throughout life. Adaptation in the nervous system in response to external stimuli requires synthesis of new gene products in order to elicit long lasting changes in processes such as development, response to injury, learning, and memory1. Information in the environment is coded in the pattern of action-potential firing, therefore gene transcription must be regulated by the pattern of neuronal firing.
    [Show full text]
  • Regulation of AMPA Receptor Subunit Glua1 Surface Expression by PAK3
    Regulation of AMPA receptor subunit GluA1 surface PNAS PLUS expression by PAK3 phosphorylation Natasha K. Hussaina, Gareth M. Thomasb, Junjie Luoa,1, and Richard L. Huganira,2 aSolomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD 21205; and bShriners Hospitals Pediatric Research Center and Department of Anatomy and Cell Biology, Temple University School of Medicine, Philadelphia, PA 19140 Contributed by Richard L. Huganir, September 17, 2015 (sent for review July 17, 2015) AMPA receptors (AMPARs) are the major excitatory receptors of the a member of a superfamily of kinases (7). The PAK protein family brain and are fundamental to synaptic plasticity, memory, and comprises six members, falling into two categories: group I (con- cognition. Dynamic recycling of AMPARs in neurons is regulated sisting of PAKs 1, 2, and 3) and group II (includes PAKs 4, 5, and through several types of posttranslational modification, including 6), which are distinguished based on sequence homology and are phosphorylation. Here, we identify a previously unidentified signal functionally regulated by distinct mechanisms. Although PAK transduction cascade that modulates phosphorylation of serine proteins differ in their specific developmental/subcellular distribu- residue 863 (S863) in the GluA1 AMPAR subunit and controls surface tion, they are primarily implicated in regulating actin cytoskeletal trafficking of GluA1 in neurons. Activation of the EphR–Ephrin sig- dynamics (8). However, the possibility that PAK signaling controls nal transduction pathway enhances S863 phosphorylation. Further, AMPAR localization and/or trafficking has not been investigated. EphB2 can interact with Zizimin1, a guanine–nucleotide exchange In this study, we identified a novel site in the GluA1 subunit of factor that activates Cdc42 and stimulates S863 phosphorylation in AMPAR (serine 863) that is rapidly phosphorylated following neurons.
    [Show full text]
  • Supplementary Table 5A
    CE2/+ f/f CA/+ CE2/+ f/f Supplementary Table 5A: Pathway enrichment analysis for differentially expressed genes between Nkx3.1 ; Pten ; Braf and Nkx3.1 ; Pten mouse prostate tumors Pathway Gene Number Enrichment Normalized p-value False Family-wise Rank Leading Edge Genes Set of genes Score enrichment Discovery Error Rate at Details in the score Rate p-value Max pathway q-value TPM2 ACTG2 MYL9 ACTA2 VCL DES MYLK ITGA1 Details TPM4 SORBS1 CALM2 ITGB5 PXN LMOD1 TNNC2 1 REACTOME_MUSCLE_CONTRACTION ... 30 0.78 2.58 0 0 0 2180 MYL1 MYH11 Details TPM2 ACTG2 MYL9 ACTA2 VCL MYLK ITGA1 2 REACTOME_SMOOTH_MUSCLE_CONTRACTION ... 20 0.81 2.39 0 0 0 1609 TPM4 SORBS1 CALM2 ITGB5 PXN LMOD1 MYH11 ITGA3 LAMB3 ITGB4 THBS1 LAMA3 CD44 ITGA6 ITGAV ITGB1 SPP1 LAMC2 ITGA1 SDC1 RELN LAMC1 FN1 ITGB5 THBS2 COL1A2 ITGA9 ITGB6 SDC2 HSPG2 TNC COL6A1 COL4A2 COL6A2 Details COL4A6 ITGA5 COL4A1 LAMA1SV2B COL5A1 3 KEGG_ECM_RECEPTOR_INTERACTION ... 59 0.63 2.32 0 0 0 3812 COL6A3 COL3A1 ITGA11 COL5A2 ITGA10 Details ACTIN1 FLNC FBLIM1 ITGB1 FLNA RSU1 FERMT2 4 REACTOME_CELLEXTRACELLULAR_MATRIX_INTERACTIONS ... 13 0.81 2.16 0 0 0.001 1193 PXN ILK ITGA3 ITGB4 THBS1 ITGA6 ITGAV ITGB1 SPP1 ITGA1 LAMC1 FN1 ITGB5 COL1A2 CDH1 BCRAR1 ITGA9 ITGB6 COL4A5 TNC COL4A2 ITGA5 COL4A1 Details LAMA1 PTPN1 FGG PECAM1 FGA FBN1 ITGA11 5 REACTOME_INTEGRIN_CELL_SURFACE_INTERACTIONS ... 62 0.57 2.09 0 0 0.002 3812 ITGA10 COL1A1 LAMA2 LAMB2 ITGA3 CCND1 LAMB3 ITGB4 THBS1 FLNB LAMA3 ACTN1 IGF1R CCND2 ITGA6 ITGAV FLNC MYL9 CAV1 ITGB1 VEGF1 PDGFRA SPP1 FLNA MAP2K1 PDGFC VCL PPP1CC MYLK LAMC2 CAPN2 CDC42 ITGA1 RELN LAMC1 AKT3 FN1 CAV2 PDGFRB ITGB5 PXN ILK PDGFB THBS2 DOCK1 COL1A2 FLT1 FIGF BCAR1 ITGA9 ITGB6 PAK3 TNC COL6A1 COL4A2 COL6A2 COL4A6 ITGA5 COL4A1 LAMA1 Details BCL2 COL5A1 PGF ZYX COL6A3 COL3A1 ITGA11 6 KEGG_FOCAL_ADHESION ..
    [Show full text]
  • The Human Gene Connectome As a Map of Short Cuts for Morbid Allele Discovery
    The human gene connectome as a map of short cuts for morbid allele discovery Yuval Itana,1, Shen-Ying Zhanga,b, Guillaume Vogta,b, Avinash Abhyankara, Melina Hermana, Patrick Nitschkec, Dror Friedd, Lluis Quintana-Murcie, Laurent Abela,b, and Jean-Laurent Casanovaa,b,f aSt. Giles Laboratory of Human Genetics of Infectious Diseases, Rockefeller Branch, The Rockefeller University, New York, NY 10065; bLaboratory of Human Genetics of Infectious Diseases, Necker Branch, Paris Descartes University, Institut National de la Santé et de la Recherche Médicale U980, Necker Medical School, 75015 Paris, France; cPlateforme Bioinformatique, Université Paris Descartes, 75116 Paris, France; dDepartment of Computer Science, Ben-Gurion University of the Negev, Beer-Sheva 84105, Israel; eUnit of Human Evolutionary Genetics, Centre National de la Recherche Scientifique, Unité de Recherche Associée 3012, Institut Pasteur, F-75015 Paris, France; and fPediatric Immunology-Hematology Unit, Necker Hospital for Sick Children, 75015 Paris, France Edited* by Bruce Beutler, University of Texas Southwestern Medical Center, Dallas, TX, and approved February 15, 2013 (received for review October 19, 2012) High-throughput genomic data reveal thousands of gene variants to detect a single mutated gene, with the other polymorphic genes per patient, and it is often difficult to determine which of these being of less interest. This goes some way to explaining why, variants underlies disease in a given individual. However, at the despite the abundance of NGS data, the discovery of disease- population level, there may be some degree of phenotypic homo- causing alleles from such data remains somewhat limited. geneity, with alterations of specific physiological pathways under- We developed the human gene connectome (HGC) to over- come this problem.
    [Show full text]
  • Membrane and Protein Interactions of the Pleckstrin Homology Domain Superfamily
    Membranes 2015, 5, 646-663; doi:10.3390/membranes5040646 OPEN ACCESS membranes ISSN 2077-0375 www.mdpi.com/journal/membranes Article Membrane and Protein Interactions of the Pleckstrin Homology Domain Superfamily Marc Lenoir 1, Irina Kufareva 2, Ruben Abagyan 2, and Michael Overduin 3,* 1 School of Cancer Sciences, Faculty of Medical and Dental Sciences, University of Birmingham, Birmingham B15 2TT, UK; E-Mail: [email protected] 2 Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California, San Diego, 9500 Gilman Drive, La Jolla, CA 92093, USA; E-Mails: [email protected] (I.K.); [email protected] (R.A.) 3 Department of Biochemistry, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, AB T6G 2H7, Canada * Author to whom correspondence should be addressed: E-Mail: [email protected]; Tel.: +1-780-492-3357; Fax: +1-780-492-0886. Academic Editor: Shiro Suetsugu Received: 15 September 2015 / Accepted: 16 October 2015 / Published: 23 October 2015 Abstract: The human genome encodes about 285 proteins that contain at least one annotated pleckstrin homology (PH) domain. As the first phosphoinositide binding module domain to be discovered, the PH domain recruits diverse protein architectures to cellular membranes. PH domains constitute one of the largest protein superfamilies, and have diverged to regulate many different signaling proteins and modules such as Dbl homology (DH) and Tec homology (TH) domains. The ligands of approximately 70 PH domains have been validated by binding assays and complexed structures, allowing meaningful extrapolation across the entire superfamily. Here the Membrane Optimal Docking Area (MODA) program is used at a genome-wide level to identify all membrane docking PH structures and map their lipid-binding determinants.
    [Show full text]