In this Administrative Supplement application, we propose to apply our targeted integration strategy to build a novel in vivo zebrafish model of microglia inflammasome signaling in neuroinflammation. Chronic neuroinflammation driven by microglia activation underlies the pathophysiology of Alzheimer?s Disease (AD) and (PD) and related dementia. a-synuclein secreted by neurons in Parkinson?s patients form aggregates that are taken up by microglia, however, our understanding of the downstream intracellular mechanisms that signal microglia inflammasome activation is incomplete. We will build on our expertise in zebrafish site directed genome engineering and mouse models of neuroinflammation to generate a zebrafish model of conditional active fyn kinase that will allow temporal and spatial induction of microglia inflammasome signaling and activation. Our model will provide an in vivo platform for phenotypic chemical screens that recapitulates the cellular complexity of neuroinflammation. The new tools we develop for microglia specific Cre expression and conditional control of inflammasome signaling will provide an important resource for zebrafish researchers investigating immune function in disease pathogenesis as well as development of translational strategies.
The central goal for this application is to generate a genetic resource for experimental investigation into neuroinflammation in neurodegenerative diseases related to Alzheimer?s. The strategies proposed employ the ability of designer nucleases to create DNA double strand breaks and subsequent site- specific integration of DNA. We propose to generate conditional gene alleles and recombinase drivers in zebrafish to investigate how microglia, the immune cells of the nervous system, contribute to neuroinflammation. The technologies developed will provide advanced tools for modeling human immune system development and disease in zebrafish.
|Schultz, Laura E; Haltom, Jeffrey A; Almeida, Maira P et al. (2018) Epigenetic regulators Rbbp4 and Hdac1 are overexpressed in a zebrafish model of RB1 embryonal brain tumor, and are required for neural progenitor survival and proliferation. Dis Model Mech 11:|