High-Content Analysis Core

ABSTRACT – CORE A Our Center proposes to study a large number of domain mutations and previously uncharacterized NDD- associated genetic variants in TRIO and SYNGAP1 using a high-throughput strategies (Project 1), to perform functional screens for newly developed chemical probes in mouse neurons (Project 1), and to generate novel iPSC-derived neuronal models (Project 2). All of these research endeavors will rely heavily upon efficient medium or high-throughput screening capabilities as well as the ability to analyze a multitude of phenotypes in dozens of conditions. Core A will provide the following services: 1) assist Projects 1 and 2 with the optimization of imaging strategies, MEA recordings, and analysis of cultured neuron systems; 2) perform high-content imaging (HCI) of neuroarchitectural phenotypes for Projects 1 and 2; 3) perform multielectrode array (MEA) analysis of neuronal network electrophysiological phenotypes for Projects 1 and 2; 4) provide instruction of Center users for HCI, MEA recordings, and analysis; 5) facilitate the offline analysis of data; 6) facilitate the storage of phenotypic data; 7) ensure quality control of data and analysis. To accomplish this work in an efficient, cost-effective, and rigorous manner, Core A will exploit standardized high-throughput workflows that unite innovative technologies with economies of scale. The Specific Aims of Core A are: Aim 1. To assess the impact of SNVs in TRIO and SYNGAP1 on neuroarchitecture and network activity in mouse neurons. Aim 2. To assess the impact of chemical modulators of GEF and GAP activity on neuroarchitecture and network activity in mouse neurons. Aim 3. To assess the impact of genetic mutations and chemical probes on human iPSC-derived neuronal models.