Systematic evaluation of extrinsic variables to improve rigor and reproducibility in biomedical research with zebrafish

About This Grant

Abstract Improvement of experimental rigor and reproducibility is critical to enhance biomedical research. Zebrafish (Danio rerio) are widely used organisms in numerous biomedical research fields, including developmental biology, genetics, neurobiology, /toxicology, and more. Zebrafish also are a key component of New Approach Methodologies (NAMs), as they provide the only in-tact vertebrate system within this category. While many aspects of zebrafish environmental factors have been standardized across laboratories, there remain many elements that have not been systematically studied to account for how they affect variability of measured biological endpoints such as behavior, organ growth, morphology, nutrient content, and xenobiotic responses. To address this data gap and enhance shared resources across our community, we propose to acquire new, state of the art equipment capable of recording, measuring, and analyzing multiple parameters simultaneously: 1) embryo incubators that monitor and control light, temperature, and humidity, 2) a multi-camera array microscope for morphology and behavior endpoints, and 3) an aquatics data system to monitor, record, and automatically adjust water quality parameters. Leveraging four independently managed zebrafish facilities at the University of Massachusetts Amherst, we aim to systematically evaluate extrinsic factors affecting biological endpoints across multiple physiological and organ systems in zebrafish at embryonic/larval (rearing media, light cycle, vessel size, population density, and feeding) and parental life stages (diet), and monitor other environmental measures (e.g. fish facility water quality parameters, temperature). Outcomes affecting developmental toxicology impacts on embryonic development, pancreas and liver structure and function, antioxidant responses, behavior, neurogenesis, craniofacial development and skeletal mineralization, will incorporate extrinsic factors to determine if they are modifiable variables. This study will lead to identification of new mechanistic bioindicators and biomarkers that can be utilized by others to improve reproducibility, rigor, and translation of findings from zebrafish to human health outcomes.