title: Estrogens Suppress a Behavioral Phenotype in Zebrafish Mutants of the Autism Risk Gene, CNTNAP2.
abstract: Autism spectrum disorders (ASDs) are a group of devastating neurodevelopmental syndromes that affect up to 1 in 68 children. Despite advances in the identification of ASD risk genes, the mechanisms underlying ASDs remain unknown. Homozygous loss-of-function mutations in Contactin Associated Protein-like 2 (CNTNAP2) are strongly linked to ASDs. Here we investigate the function of Cntnap2 and undertake pharmacological screens to identify phenotypic suppressors. We find that zebrafish cntnap2 mutants display GABAergic deficits, particularly in the forebrain, and sensitivity to drug-induced seizures. High-throughput behavioral profiling identifies nighttime hyperactivity in cntnap2 mutants, while pharmacological testing reveals dysregulation of GABAergic and glutamatergic systems. Finally, we find that estrogen receptor agonists elicit a behavioral fingerprint anti-correlative to that of cntnap2 mutants and show that the phytoestrogen biochanin A specifically reverses the mutant behavioral phenotype. These results identify estrogenic compounds as phenotypic suppressors and illuminate novel pharmacological pathways with relevance to autism.
authors.affiliation: Child Study Center, Yale School of Medicine, New Haven, CT 06510, USA; Program on Neurogenetics, Yale School of Medicine, New Haven, CT 06510, USA.
authors.forename: Ellen J
authors.initials: EJ
authors.lastname: Hoffman
authors.affiliation: Department of Cell and Developmental Biology, University College London, Gower Street, London WC1E 6BT, UK.
authors.forename: Katherine J
authors.initials: KJ
authors.lastname: Turner
authors.affiliation: Child Study Center, Yale School of Medicine, New Haven, CT 06510, USA; Program on Neurogenetics, Yale School of Medicine, New Haven, CT 06510, USA.
authors.forename: Joseph M
authors.initials: JM
authors.lastname: Fernandez
authors.affiliation: Department of Genetics, Yale University School of Medicine, New Haven, CT 06510, USA; Department of Biochemistry, Boston University School of Medicine, Boston, MA 02118, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: Cifuentes
authors.affiliation: Department of Cell and Developmental Biology, University College London, Gower Street, London WC1E 6BT, UK.
authors.forename: Marcus
authors.initials: M
authors.lastname: Ghosh
authors.affiliation: Child Study Center, Yale School of Medicine, New Haven, CT 06510, USA; Program on Neurogenetics, Yale School of Medicine, New Haven, CT 06510, USA.
authors.forename: Sundas
authors.initials: S
authors.lastname: Ijaz
authors.affiliation: Department of Cell and Developmental Biology, University of Pennsylvania, Perelman School of Medicine, Philadelphia, PA 19104, USA; Department of Biology, Haverford College, Haverford, PA 19041, USA.
authors.forename: Roshan A
authors.initials: RA
authors.lastname: Jain
authors.affiliation: Department Genes - Circuits - Behavior, Max Planck Institute of Neurobiology, Am Klopferspitz 18, 82152 Martinsried, Germany.
authors.forename: Fumi
authors.initials: F
authors.lastname: Kubo
authors.affiliation: Center for Autism Research and Treatment, Semel Institute for Neuroscience and Human Behavior, Department of Psychiatry, University of California, Los Angeles, Los Angeles, CA 90095, USA; Department of Biology, The University of Texas at Tyler, Tyler, TX 75799, USA.
authors.forename: Brent R
authors.initials: BR
authors.lastname: Bill
authors.affiliation: Department Genes - Circuits - Behavior, Max Planck Institute of Neurobiology, Am Klopferspitz 18, 82152 Martinsried, Germany.
authors.forename: Herwig
authors.initials: H
authors.lastname: Baier
authors.affiliation: Department of Cell and Developmental Biology, University of Pennsylvania, Perelman School of Medicine, Philadelphia, PA 19104, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Granato
authors.affiliation: Department of Biological Sciences, Smith College, Northampton, MA 01063, USA.
authors.forename: Michael J F
authors.initials: MJ
authors.lastname: Barresi
authors.affiliation: Department of Cell and Developmental Biology, University College London, Gower Street, London WC1E 6BT, UK.
authors.forename: Stephen W
authors.initials: SW
authors.lastname: Wilson
authors.affiliation: Department of Cell and Developmental Biology, University College London, Gower Street, London WC1E 6BT, UK. Electronic address: j.rihel@ucl.ac.uk.
authors.forename: Jason
authors.initials: J
authors.lastname: Rihel
authors.affiliation: Child Study Center, Yale School of Medicine, New Haven, CT 06510, USA; Program on Neurogenetics, Yale School of Medicine, New Haven, CT 06510, USA; Department of Genetics, Yale University School of Medicine, New Haven, CT 06510, USA; Department of Psychiatry, University of California, San Francisco, San Francisco, CA 94143, USA. Electronic address: matthew.state@ucsf.edu.
authors.forename: Matthew W
authors.initials: MW
authors.lastname: State
authors.affiliation: Department of Genetics, Yale University School of Medicine, New Haven, CT 06510, USA. Electronic address: antonio.giraldez@yale.edu.
authors.forename: Antonio J
authors.initials: AJ
authors.lastname: Giraldez
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001321:Autistic Disorder / Q000188:drug therapy* / Q000235:genetics; D004195:Disease Models, Animal; D004967:Estrogens / Q000494:pharmacology* / Q000627:therapeutic use; D005786:Gene Expression Regulation / Q000187:drug effects* / Q000235:genetics*; D019833:Genistein / Q000494:pharmacology; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007814:Larva; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics*; D009043:Motor Activity / Q000187:drug effects / Q000235:genetics; D009154:Mutation / Q000235:genetics*; D009419:Nerve Tissue Proteins / Q000235:genetics*; D010641:Phenotype; D048789:Phytoestrogens / Q000494:pharmacology; D011619:Psychotropic Drugs / Q000494:pharmacology / Q000627:therapeutic use; D012640:Seizures / Q000188:drug therapy / Q000235:genetics; D020922:Sleep-Wake Transition Disorders / Q000188:drug therapy / Q000235:genetics; D050599:Vesicular Glutamate Transport Protein 2 / Q000235:genetics / Q000378:metabolism; D015027:Zebrafish
chemical_list: C403049:CNTNAP2 protein, human; D004967:Estrogens; D008164:Luminescent Proteins; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D048789:Phytoestrogens; D011619:Psychotropic Drugs; D050599:Vesicular Glutamate Transport Protein 2; C413663:fluorescent protein 583; D049452:Green Fluorescent Proteins; D019833:Genistein; C004541:biochanin A
references.citation: J Pharmacol Sci. 2010;112(3):282-9
references.pmid: 20168048
references.citation: N Engl J Med. 2006 Mar 30;354(13):1370-7
references.pmid: 16571880
references.citation: Neuroscience. 2004;126(3):591-8
references.pmid: 15183509
references.citation: Neuron. 2015 Sep 23;87(6):1215-33
references.pmid: 26402605
references.citation: Front Neuroendocrinol. 2014 Aug;35(3):255-71
references.pmid: 24705124
references.citation: Cell. 2013 Nov 21;155(5):1008-21
references.pmid: 24267887
references.citation: Nature. 2014 Nov 13;515(7526):216-21
references.pmid: 25363768
references.citation: J Cell Biol. 2003 Sep 15;162(6):1149-60
references.pmid: 12963709
references.citation: Psychoneuroendocrinology. 2009 Dec;34 Suppl 1:S130-42
references.pmid: 19596521
references.citation: Genes Brain Behav. 2003 Oct;2(5):255-67
references.pmid: 14606691
references.citation: Nature. 2014 Nov 13;515(7526):209-15
references.pmid: 25363760
references.citation: Autism Res. 2013 Apr;6(2):69-77
references.pmid: 23441094
references.citation: Science. 2010 Jan 15;327(5963):348-51
references.pmid: 20075256
references.citation: J Neurosci. 2006 Dec 20;26(51):13400-10
references.pmid: 17182791
references.citation: J Neurosci. 2000 Jan 15;20(2):709-21
references.pmid: 10632600
references.citation: Trends Cell Biol. 2013 Dec;23(12):584-6
references.pmid: 24275383
references.citation: Neuroscience. 2005;131(3):759-68
references.pmid: 15730879
references.citation: Genome Res. 2003 Apr;13(4):533-43
references.pmid: 12670995
references.citation: Proc Natl Acad Sci U S A. 2011 Jan 18;108(3):1164-9
references.pmid: 21199947
references.citation: Cell. 2013 Nov 21;155(5):997-1007
references.pmid: 24267886
references.citation: PLoS Biol. 2011 Jun;9(6):e1001081
references.pmid: 21695109
references.citation: Nat Neurosci. 2005 Aug;8(8):1059-68
references.pmid: 16007083
references.citation: N Engl J Med. 2002 Aug 1;347(5):314-21
references.pmid: 12151468
references.citation: Cell. 2011 Sep 30;147(1):235-46
references.pmid: 21962519
references.citation: Methods Cell Biol. 2011;105:517-24
references.pmid: 21951545
references.citation: Curr Genomics. 2006;7(8):497-508
references.pmid: 18369406
references.citation: Am J Hum Genet. 2014 Mar 6;94(3):415-25
references.pmid: 24581740