Reactive gliosis in adult zebrafish telencephalon following daily nanoplastic consumption

Abstract

Background: Polystyrene nanoplastics are one of the most pervasive plastic contaminates and have been shown to infiltrate multiple mediums that are ingested by humans including air, food and water. Additionally, polystyrene nanoparticles have been shown to cross the blood brain barrier (BBB), the brain's first defense against pathogens and toxins. Following insult, the neuronal immune network enters a pro-inflammatory state referred to as reactive gliosis. Reactive gliosis is characterized by the recruitment of microglia and astroglia, the resident immune cells of the central nervous system (CNS), into a pro-inflammatory state that leads to the release of cytokines and the creation of a glial scar. This cascade of morphological and chemical changes in the CNS following prolonged insult can induce significant damage to the brain. Experiments in rodent models and cell culture indicate that exposure to plastic nanoparticles can induce inflammatory processes in the brain. This has been relatively underexplored in the zebrafish model system. In the current study we investigate the hypothesis that daily exposure to nanoplastics via the food chain can induce a chronic inflammatory state in the brain that may lead to permanent brain damage. The purpose of the current study is to investigate reactive gliosis in the telencephalon of adult zebrafish following the consumption of polystyrene-contaminated food. Methods: One thousand brine shrimp ( Artemia) per zebrafish were collected and incubated in artificial sea water (ASW) containing 44 nanometer (nm) fluorescent polystyrene (nPS) at a concentration of 0.15 milligrams (mg) nPS/50 milliliters (mL) ASW for 24 hours. Zebrafish were fed polystyrene-enriched brine shrimp once per day for fourteen consecutive days. On day fifteen, the telencephalon was removed and homogenized for western blot analysis. Reactive microglia were measured using the monoclonal 4C4 antibody which has been found to label reactive microglia. Reactive astrogliosis was quantified by measuring glial fibrillary acidic protein (GFAP), a marker for reactive astrocytes. Results: Confocal fluorescent imaging and spectrophotometry showed that Artemia were enriched with red-fluorescently tagged polystyrene nanoplastics. Additionally, the quantity of nanoplastic within one thousand shrimp was estimated to be 5 +/- 1.9 micrograms. There was an observed increase in 4C4 normalized to actin. Increases in GFAP were also detected. These data indicate that reactive gliosis by microglia and astroglia was enhanced in the zebrafish telencephalon after consuming polystyrene-contaminated food for fourteen consecutive days. Funding is supported by the Georgia Southern Department of Biology and Mercer University College of Medicine. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.