Autism Spectrum Disorders (ASDs) are currently estimated to impact 1-2.6% of children world-wide, representing a steep rise in ASD prevalence with annual costs to the United States alone calculated at $126 billion (2012). Because known therapies are less effective with increasing age of diagnosis, addressing outstanding questions about ASD etiology is a research priority. In the more common mammalian models however, embryonic stages are inaccessible therefore embryogenesis presents a major gap in our understanding of ASD etiology. To address this gap, we propose to generate zebrafish ASD models to focus explicitly on functional consequences in embryos of mutations known to cause ASD. Rather than investigate social behaviors typically used to define ASD, we focus on internal phenotypes (endophenotypes) of neuroanatomy and physiology. Following this strategy, our preliminary data from morpholino knockdown experiments demonstrate common phenotypes when either of two distinct ASD-linked genes, SHANK3 and SYNGAP1, is knocked down in zebrafish. Common phenotypes include developmental delay and seizure-like behaviors. These seizure-like behaviors are likely explained by dramatic reductions in the numbers of inhibitory GABAergic neurons in both morphant models. Developmental delay, seizures, and reduced markers of GABAergic signaling are also characteristic of individuals with ASD. To follow up on these preliminary studies we propose to generate stable gene knock-outs of zebrafish shank3 and syngap1. By creating and analyzing stable mutant lines with respect to the development of GABAergic brain circuits, our goal is to determine the developmental mechanisms that underlie GABA deficits. In the long-term, these zebrafish ASD models can also serve as the basis for the discovery of therapeutic targets and environmental risk factors.
Our basic research proposal is relevant to public health because it generates zebrafish models of two 'single gene'mutations linked to Autism Spectrum Disorder (ASD). Stable shank3 and syngap1 mutants will contribute to our understanding of neural circuit disruptions that characterize Autism Spectrum Disorders and, in the long-term, provide animal models for drug screens to identify therapeutic strategies and environmental risk factors. Such research is a relevant part of the NIH's mission because it will provide fundamental knowledge and develop key animal models to help with the management of a human disorder that costs billions of dollars annually and affects millions of people.
|Kozol, Robert A; Cukier, Holly N; Zou, Bing et al. (2015) Two knockdown models of the autism genes SYNGAP1 and SHANK3 in zebrafish produce similar behavioral phenotypes associated with embryonic disruptions of brain morphogenesis. Hum Mol Genet 24:4006-23|