The Influence of Lead on Different Proteins in Gill Cells From the Freshwater Bivalve, Corbicula fluminea, From Defense to Repair Biomarkers

Abstract

The objective of this study was to evaluate the influence of lead (Pb) on regulatory proteins linked to mechanisms of animal adaptation to polluted environments (using in vivo and in vitro tests) and to validate the in vitro assay as a tool for environmental assessment. Specimens of the bivalve Corbicula fluminea were exposed to nominal concentrations of Pb 5 mg l(-1) for 96 h. Isolated gill cells were exposed to three concentrations (1, 10, and 100 μM) for 5 h. Metal toxicity was evaluated by cell viability (trypan blue exclusion). We also analyzed Na+/K+ adenosine triphosphatase (ATPase) and carbonic anhydrase activity. Additionally, the multixenobiotic-resistance (MXR) phenotype was evaluated by the accumulation of rhodamine B (RB). Immunolabeling was used to quantify the expression of P-glycoproteins (C219) and proteins involved in ion transport, water movement, and cellular repair using antibodies against Na+/K+ ATPase, aquaporin 1, and heat-shock protein 70 (Hsp70). Pb was shown to be toxic in both in vivo and in vitro tests, in which cellular viability significantly decreased by approximately 25%. Cellular viability in the in vivo assays was determined by gill cell isolation after the entire animal was exposed to Pb. We observed that Na+/K+ ATPase activity was inhibited by 70%. Also, the expression of the MXR phenotype significantly increased in our in vivo tests. A statistically significant difference was observed in the expression of all proteins in the in vitro assays, whereas only Hsp70 increased in vivo. Employing these analyses, we could validate the sensitivity of the in vitro tests and can propose our in vitro model as a possible tool for environmental assessment.

Cytotoxicity of the association of pesticides Roundup Transorb® and Furadan 350 SC® on the zebrafish cell line, ZF-L

“…For this, the cells in plates were fixed in formaldehyde (4%) and stained with monoclonal antibody C219 (against Pgp) (SIGNE-COVANCE, Kansas City, USA) and then with the fluorescent secondary antibody TRITC (Sigma Aldrich, St Louis -USA) which binds to the primary antibody (Rocha and Souza, 2012). In order to show the efficiency of technique, we performed three samples without primary antibody, as negative control.…”

Multi-Biomarker Responses of Asian Clam Corbicula fluminea (Bivalvia, Corbiculidea) to Cadmium and Microplastics Pollutants

“…The freshwater bivalve Asian clams (C. fluminea) have successfully been used to monitor pollution levels in aquatic ecosystems, with in situ toxicity tests (e.g., nanoparticles, toxic trace elements) [8,9], and to study the toxicological effects of MPs in the laboratory [10]. This benthic mollusc has also been regularly adopted as a model due to its strong ability to bioaccumulate metals [8,11,12], and among the tested metals, cadmium (Cd) is the most noxious for freshwater organisms, including C. fluminea [13].…”

Contaminants from dredged sediments alter the transcriptome of Manila clam and induce shifts in microbiota composition

“…ABC transporters are a family of proteins that play a key role in cellular detoxification, facilitating the transport of various substances out of cells. Several studies showed that the ABC protein family can be involved in the excretion of metal ions [ 18 – 21 ]. Shi and Xiang [ 22 ] showed that Cu bioaccumulation in Crassostrea angulata was significantly higher after interference with an ABC gene, concluding that the expression levels of this gene affect the accumulation of copper in oysters.…”