Glutathione, Glutathione-Related Enzymes, and Catalase Activities in the Earthworm Eisenia fetida andrei

Abstract

The aim of this work was to provide basic data on the antioxidant defences in the annelid Eisenia fetida andrei (E. f. a.). Methods for measurement of three antioxidant enzymes-catalase (CAT), glutathione peroxidase (GPX), and glutathione reductase (GR)-and of glutathione-S-transferase (GST) were optimized. GPX activity differed according to the substrate used: cumene hydroperoxide (CUOOH) or hydrogen peroxide (H2O2). The effects on the enzyme activities of storage up to 2 months at -80 degrees C, -20 degrees C, and +4 degrees C were evaluated. The subcellular distribution (in cytosol, mitochondrial, and microsomal fractions) was examined. The properties and subcellular distribution of the enzymes and glutathione were also characterized in dissected tissues and body fluids. The GR activity decreased at -80 degrees C and was the only one not stable at this temperature. The four enzymes were localized mainly in the cytosolic fraction. CAT distribution was unusual as it was not associated with peroxisomes, its properties being consistent with a catalase-peroxidase, rather than a true catalase. However, this result could also be an artifact linked to the use of an inappropriate method to obtain the fractions. Our observations indicate the presence of a distinct cytosolic selenium-dependent GPX (Se-GPX), and of a possible microsomal Se-GPX. A strong non-Se-GPX activity was measured in the CF and CL, which could be linked to the peroxidase activity of fetidins secreted by coelomocytes and with the ROS production of these cells. This study seems to indicate that E. f. a. is well equipped for the metabolism of electrophilic and pro-oxidants through glutathione.

Antioxidant enzyme activities and markers of oxidative stress in the life cycle of earthworm,Eudrilus eugeniae

“…The juveniles showed higher SOD activity compared to other groups. Considering the fact that metabolic rate is higher in juvenile than in adult earthworms, the former require higher SOD activities to protect against increased amount of ROS (Saint-Denis et al 1998).…”

“…The effects of metals on the CAT and glutathione peroxidase (GPX) activities in Eisenia fetida andrei have also been studied by Labrot et al (1996) both in vivo and in vitro. A study on the activities of the CAT, GPX and glutathione reductase (GR) involved in the antioxidant defences of Eisenia fetida andrei has been conducted by Saint-Denis et al (1998). These results suggest that the antioxidant system of earthworms should be interesting to study as a potential biomarker.…”

Toxicity, bioaccumulation and metabolism of pesticides in the earthworm

“…wt. of the CDNB-active GST to be 46.2 kDa 146) with an optimal pH of 7.5 for activity 153) in Eisenia sp. GST isoforms in the post-mitochondrial fraction of E. fetida, partially purified by GSH-affinity chromatography, were subjected to SDS-PAGE, and four subunits with mol.…”

Survival, growth, detoxifying and antioxidative responses of earthworms (Eisenia fetida) exposed to soils with industrial DDT contamination

“…CAT activity was measured as described in Claiborne [24] and Saint-Denis et al [25,26]. The variation of absorbance at 240 nm due to the dismutation of H 2 O 2 was measured.…”

“…GST activity was measured using the method described in Saint-Denis et al [25,26]. The assay was conducted by monitoring the appearance of the conjugated complex of CDNB and GSH at 340 nm over time.…”