A rotenone organotypic whole hemisphere slice model of mitochondrial abnormalities in the neonatal brain

A rotenone organotypic whole hemisphere slice model of mitochondrial abnormalities in the neonatal brain

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Abstract Mitochondrial abnormalities underscore a variety of lagoon (snes neurologic injuries and diseases and are well-studied in adult populations.Clinical studies identify critical roles of mitochondria in a wide range of developmental brain injuries, but models that capture mitochondrial abnormalities in systems representative of the neonatal brain environment are lacking.Here, we develop an organotypic whole-hemisphere (OWH) brain slice model of mitochondrial dysfunction in the neonatal brain.We extended the utility of complex I inhibitor rotenone (ROT), canonically used in models of adult neurodegenerative diseases, to inflict mitochondrial damage in OWH slices from term-equivalent rats.

We quantified whole-slice health over 6 days of exposure for a range of doses represented in ROT literature.We identified 50 nM ROT as a suitable exposure level for OWH slices to inflict injury without compromising viability.At the selected exposure level, we confirmed exposure- and time-dependent mitochondrial responses showing differences in mitochondrial fluorescence and nuclear localization using MitoTracker imaging in live OWH slices and dysregulated mitochondrial markers via RT-qPCR screening.We leveraged the regional structures present in OWH slices to quantify cell density and cell death in the how to rebuild a 1963 corvette front suspension cortex and the midbrain regions, observing higher susceptibilities to damage in the midbrain as a function of exposure and culture time.

We supplemented these findings with analysis of microglia and mature neurons showing time-, region-, and exposure-dependent differences in microglial responses.We demonstrated changes in tissue microstructure as a function of region, culture time, and exposure level using live-video epifluorescence microscopy of extracellularly diffusing nanoparticle probes in live OWH slices.Our results highlight severity-, time-, and region-dependent responses and establish a complimentary model system of mitochondrial abnormalities for high-throughput or live-tissue experimental needs.