, Volume 5, Issue 1, pp 92-101

Expression and Characterization of European Sea Bass (Dicentrarchus labrax) Somatolactin: Assessment of In Vivo Metabolic Effects

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Abstract

The complementary DNA coding for European sea bass somatolactin was expressed in the pET-3a bacteria expression vector. The recombinant somatolactin (rbSL) was purified by size exclusion chromatography, and 95% of the protein remained in the oxidized form with negligible aggregation over prolonged cold storage. The identity of the recombinant protein was demonstrated by Western blotting with a rabbit polyclonal antibody against gilthead sea bream somatolactin. The same antibody was utilized in a radioimmunoassay procedure, using rbSL as standard and radioiodinated tracer. Curve displacements of pituitary and plasma samples paralleled the rbSL standard, and the midrange of the assay (8 ng/ml) was low enough to measure in a consistent manner the circulating SL concentration. To assess biological activity a single dose of rbSL (0.1 µg/g of body mass) was administered to juvenile gilthead sea bream by intraperitioneal injection. In comparison with saline-treated fish, rbSL did not modify the circulating amount of insulin-like growth factor I, whereas a 50% increase was found with the same dose of recombinant trout growth hormone (rtGH). Hormone treatment did not modify nitrogen-ammonia excretion, but both rbSL and rtGH increased carbon dioxide output and oxygen uptake, which in turn decreased the respiratory quotient (CO2 output per O2 uptake). This pattern of gas exchange suggests the enhancement of lipid catabolism, which is consistent with the observation that both hormones were able to inhibit the hepatic activity of acetyl–coenzyme A carboxylase. These new insights provide direct evidence for the involvement of fish somatolactin in energy homeostasis, which may serve to maintain the lipolytic tonus in different physiologic states.