Redox proteomic analysis of mytilus edulis gills: effects of the pharmaceutical diclofenac on a non‐target organism

Abstract

Veterinary and human pharmaceuticals are an emerging category of chemical pollutants with potential to cause serious toxicity to non-target organisms. Filter-feeding aquatic organisms such as mussels are especially threatened. In this study, the blue mussel, Mytilus edulis, was exposed to two doses (0.2 mg/L and 1 mg/L) of the anti-inflammatory diclofenac. Effects on the gill, the principal feeding organ of mussels, were investigated. It was noted that, while no effect was evident on gill glutathione transferase or catalase activities, there was a tissue-specific increase in glutathione reductase activity and reduction in total protein thiol groups. Two dimensional electrophoresis was performed and some affected proteins identified by in-gel tryptic digestion and peptide mass fingerprinting. Of these, four unique proteins (caspase 3/7-4, heat-shock cognate protein 70, a predicted enolase-like protein, arginine kinase) were found to be oxidized whilst eight unique proteins (β-tubulin, actin, isocitrate dehydrogenase, arginine kinase, heavy metal-binding HIP, cytosolic malate dehydrogenase, proteasome subunit alpha type 2, Mg: bb02e05 (glyceraldehyde-3-phosphate dehydrogenase) and superoxide dismutase) were found to have altered abundance. In addition, bioinformatic analysis suggested putative identities for six hypothetical proteins which either were oxidized or decreased in abundance. These were; 78 kDa glucose-regulated protein precursor, α-enolase, calreticulin, mitochondrial H + -ATPase, palmitoyl protein thioesterase 1 and initiation factor 5a. It is concluded that diclofenac causes significant oxidative stress to gills and that this affects key structural, metabolic and stress-response proteins.

Evaluation of Different Functional Antioxidant Groups and Protein Responses in Asian Seabass (Lates calcarifer Bloch, 1970) as an Early Indication of Fish Health in Aquaculture Farms.

“…The level of GST response was higher compared to GR. This agrees with the previous reported studies (Song et al, 2012;Jaafar et al, 2015;Gianng et al, 2018).…”

“…All samples were extracted using homogenizer (ULTRA TURREX) in 10 mM Tris -HCL buffer (pH 7.2) and the post-mitochondrial supernatants (PMS) was obtained after centrifugation at 15000 X g and 4ºC for one hour (Jaafar et al, 2015). Protein mass determination was done for all samples following Bradford assay using Bovine Serum Albumin (BSA) as a standard (Bradford, 1976).…”

“…The replicates concept for the analysis of protein damages, thiols (-SH-) and carbonyls (-CH-) followed the concept of antioxidant activity with the same sample from biological replicates used for antioxidant activity and followed with 2 gels replicates as technical replicates for each sample. All samples were extracted using homogeniser (ULTRA TURREX) in 10 mM Tris -HCL buffer (pH 7.2) and the postmithochondrial supernatants (PMS) was obtained after centrifugation at 15000 X g and 4ºC for one hour (Jaafar et al, 2015). Protein mass determination was done for all samples following Bradford assay using Bovine Serum Albumin (BSA) as a standard (Bradford, 1976).…”

Exposure of ELF-EMF and RF-EMF Increase the Rate of Glucose Transport and TCA Cycle in Budding Yeast

“…The IDP3 gene encodes peroxisomal NADP-dependent isocitrate dehydrogenase (ICDH), which catalyzes the oxidation of isocitrate to form alpha-ketoglutarate and NAD(P)H (Lu and McAlister-Henn, 2010). Environmental stresses, such as nutrient stress, temperature alteration, and tobacco smoking, have been shown to modulate ICDH activity (Konga et al, 2009; Jaafar et al, 2015). NAD(P)H is an effective scavenger of free radicals and hydrogen peroxide; thus ICDH might also act as an indirect antioxidant by providing NAD(P)H, which is required in corneal defense against oxidative damage (Lassen et al, 2008).…”

Circadian and Circatidal Rhythms of Protein Abundance in the Intertidal Mussel Mytilus californianus

“…individual, but studies have found that Mytilus californianus is capable of filtering at a rate of 50 -3000 mL/min (Bayne et al, 1976). This unique and important organ, serving a dual purpose for bivalve feeding and respiration, makes it a common focal tissue for studies of Mytilus stress physiology (Dahlhoff et al, 2002;Jaafar et al, 2015;Letendre et al, 2009;McDonagh et al, 2006;Rocher et al, 2015;Tomanek and Zuzow, 2010). Furthermore, the direct interaction between the gill tissue of sessile bivalves and their environment leaves them acutely exposed to pollutants, making this organ of great interest for research supporting ecotoxicological monitoring programs (Gómez-Mendikute et al, 2005).…”