Abstract
The present study was designed to evaluate the responsiveness of modulation of glial fibrillary acidic protein (GFAP) content and its fragmentation in the snake brain as a biomarker of local industrial pollution of aquatic ecosystems. Despite GFAP being a well known cytoskeleton marker of astrocytes’ reactivity in the brain of vertebrates, its expression in the snake brain remains insufficiently described. The GFAP expression and its fragmentation were detected using the immunoblot method in the snake brain. ROS level was determined with dichlorofluorescein diacetate fluorescence. The content of the glial fibrillary acidic protein (GFAP) of filament (cytoskeleton) and soluble (cytosol) fractions in the brain of dice snake Natrix tessellata from three ecosystems with different rates of industrial pollution were studied (two polluted and one clean control site). Characteristic increase in GFAP fragmentation was noted for the snakes from both the researched polluted sites. Significant increase in the content of the GFAP cleaved polypeptide fragments induced by industrial pollution exposure was confirmed in the snakes’ brains. Meaningful GFAP fragmentation was determined in snake brain astrocytes as an increase in cleaved fragments of 47–35 kDa molecular weight for both soluble and cytoskeletal GFAP fractions. We found significant abnormality in the ratio of the GFAP soluble fraction to the cytoskeletal one in contaminant-exposed dice snakes. It should testify to significant metabolic disturbance in nerve cells of the dice snakes. Furthermore, growth of reactive oxygen species level as the main cause of oxidative stress was determined in brains of the snakes exposed to environmental toxicity. Thus, astrocyte cytoskeleton disorders are associated with pollutant-induced redox imbalance in the snake brain. Despite the limited data on glial cell biology in the reptilian brain, the observed results prove that snake astrocytes can respond to the environmental toxicity using typical astroglial response. The presented results evidence that monitoring of molecular characteristics of glial cytoskeleton in dice snakes could be used as reliable biomarker of neurotoxicity and adverse effects of industrial pollution. Further studies are required to elucidate the role of astrocyte cytoskeleton in the response against neurotoxic contaminants.
Highlights
Nowadays, practically all ecosystems are exposed to adverse effects of toxic industrial-derived compounds
The samples of every brain homogenate in volume 100 μL were mixed with 1 mL of the aforementioned buffer and 5 μL of 10 μM 2′,7′-dichlorofluorescein diacetate (DCFDA)
Our findings have shown meaningful changes in cytosolic soluble glial fibrillary acidic protein (GFAP) fraction (s-GFAP) in animals exposed to environmental toxicants as compared to the chosen control group (Fig. 5)
Summary
Practically all ecosystems are exposed to adverse effects of toxic industrial-derived compounds. Biosystems demonstrate different responses to the effect of an ecotoxicant that could be observed throughout large geographical areas (Barker & Tingey, 1992; Sanderfoot & Holloway, 2017; Cantonati et al, 2020). As a rule, pronounced multiple ecotoxic effects are accompanied by different deviations of animal behaviour, population parameters and species abundance (Newman & Clements, 2007; Peterson et al, 2017; Lieshchova et al, 2018, 2019). Ecotoxicants cause changes in mortality, reproduction, community characteristics and other parameters of biosystems at any above species level (Clements & Rohr, 2009; Van de Perre et al, 2016; Schäfer & Bundschuh, 2018). Biochemical processes and molecular interactions are a background of modulation at all biosystem levels including consequent alterations in populations and ecosystem structure
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