Variations on stress defences and metallothionein levels in the Senegal sole, Solea senegalensis, during early larval stages

Abstract

The biochemical status of antioxidant defences of larvae from the commercial, benthic fish, Solea senegalensis, were studied over a period of 28 days from hatching. The parameters studied were: catalase (EC 1.11.1.6), total glutation peroxidase activity (sum of the selenium dependent and independent forms) (EC 1.6.4.2), DT-diaphorase (EC 1.6.99.2), and as an indication of conjugative detoxifying metabolism, the enzyme glutathione S-transferase (GST; EC 2.5.1.18). Oxidative damage was evaluated by the formation of malondialdehyde (MDA) as an indication of lipid peroxidation (LP). Metallothionein (MT) levels were measured by differential pulse polarography and stress proteins (HSP 70 and HSP 60) were detected by immunoblotting, using commercial antibodies. The presence of catalytic activities was observed from hatching day and tended to increase during larval development. Significant changes were seen in most enzymes at day 3, when the larvae finished their endogenous feeding and started to feed on rotifers. DT-diaphorase activities were similar in both NADH and NADPH dependent forms and, in turn, were about 10-times lower than their correspondent reductases. LP sharply increased at 19 and 28 days post-hatch (dph), suggesting a saturation of the antioxidant defences during metamorphosis. MT showed high values during the endogenous phase and the lowest value corresponded to day 3, when the egg-yolksac was fully reabsorbed; a steady-state value was reached thereafter. Stress proteins were present at all stages, showing distinctiveness in the molecular weight of HSP60 at hatching day and 1 dph. Quantitatively, both forms were significantly elevated at 3 dph, with their ratio (HSP70/HSP60) decreasing over time