Estrogens Suppress a Behavioral Phenotype in Zebrafish Mutants of the Autism Risk Gene, CNTNAP2

Ellen J Hoffman; Katherine J Turner; Joseph M Fernandez; Daniel Cifuentes; Marcus Ghosh; Sundas Ijaz; Roshan A Jain; Fumi Kubo; Brent R Bill; Herwig Baier; Michael Granato; Michael J F Barresi; Stephen W Wilson; Jason Rihel; Matthew W State; Antonio J Giraldez

doi:10.1016/j.neuron.2015.12.039

Estrogens Suppress a Behavioral Phenotype in Zebrafish Mutants of the Autism Risk Gene, CNTNAP2

Neuron. 2016 Feb 17;89(4):725-33. doi: 10.1016/j.neuron.2015.12.039. Epub 2016 Jan 28.

Authors

Ellen J Hoffman¹, Katherine J Turner², Joseph M Fernandez¹, Daniel Cifuentes³, Marcus Ghosh², Sundas Ijaz¹, Roshan A Jain⁴, Fumi Kubo⁵, Brent R Bill⁶, Herwig Baier⁵, Michael Granato⁷, Michael J F Barresi⁸, Stephen W Wilson², Jason Rihel⁹, Matthew W State¹⁰, Antonio J Giraldez¹¹

Affiliations

¹ Child Study Center, Yale School of Medicine, New Haven, CT 06510, USA; Program on Neurogenetics, Yale School of Medicine, New Haven, CT 06510, USA.
² Department of Cell and Developmental Biology, University College London, Gower Street, London WC1E 6BT, UK.
³ 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.
⁴ 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.
⁵ Department Genes - Circuits - Behavior, Max Planck Institute of Neurobiology, Am Klopferspitz 18, 82152 Martinsried, Germany.
⁶ 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.
⁷ Department of Cell and Developmental Biology, University of Pennsylvania, Perelman School of Medicine, Philadelphia, PA 19104, USA.
⁸ Department of Biological Sciences, Smith College, Northampton, MA 01063, USA.
⁹ Department of Cell and Developmental Biology, University College London, Gower Street, London WC1E 6BT, UK. Electronic address: j.rihel@ucl.ac.uk.
¹⁰ 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.
¹¹ Department of Genetics, Yale University School of Medicine, New Haven, CT 06510, USA. Electronic address: antonio.giraldez@yale.edu.

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.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Animals, Genetically Modified
Autistic Disorder / drug therapy*
Autistic Disorder / genetics
Disease Models, Animal
Estrogens / pharmacology*
Estrogens / therapeutic use
Gene Expression Regulation / drug effects*
Gene Expression Regulation / genetics*
Genistein / pharmacology
Green Fluorescent Proteins / genetics
Green Fluorescent Proteins / metabolism
Humans
Larva
Luminescent Proteins / genetics
Luminescent Proteins / metabolism
Membrane Proteins / genetics*
Motor Activity / drug effects
Motor Activity / genetics
Mutation / genetics*
Nerve Tissue Proteins / genetics*
Phenotype
Phytoestrogens / pharmacology
Psychotropic Drugs / pharmacology
Psychotropic Drugs / therapeutic use
Seizures / drug therapy
Seizures / genetics
Sleep-Wake Transition Disorders / drug therapy
Sleep-Wake Transition Disorders / genetics
Vesicular Glutamate Transport Protein 2 / genetics
Vesicular Glutamate Transport Protein 2 / metabolism
Zebrafish

Substances

CNTNAP2 protein, human
Estrogens
Luminescent Proteins
Membrane Proteins
Nerve Tissue Proteins
Phytoestrogens
Psychotropic Drugs
Vesicular Glutamate Transport Protein 2
fluorescent protein 583
Green Fluorescent Proteins
Genistein
biochanin A

Abstract

Publication types

MeSH terms

Substances

Grants and funding