Establishment of a time-resolved fluoroimmunoassay for measuring plasma insulin-like growth factor I (IGF-I) in fish: effect of fasting on plasma concentrations and tissue mRNA expression of IGF-I and growth hormone (GH) in channel catfish ( Ictalurus punctatus)

Abstract

A time-resolved fluoroimmunoassay (TR-FIA) was established and validated that allows for the determination of plasma concentrations of insulin-like growth factor I (IGF-I) in three domestically cultured fishes: channel catfish ( Ictalurus punctatus), hybrid striped bass ( Morone chrysops × M. saxatilis), and rainbow trout ( Oncorhynchus mykiss). Sensitivity of the assay was 0.20 ng/ml. Intra- and inter-assay coefficients of variation (CV) were <7 and <12%, respectively. Serial dilutions of plasma from each species were parallel to the standard curve. Recovery of IGF-I from spiked plasma samples was >90% for all three species of fishes. The IGF-I TR-FIA was biologically validated via its use to determine the effect of fasting on circulating IGF-I levels in channel catfish. Fasting-induced changes in plasma growth hormone (GH), hepatic IGF-I mRNA expression, and pituitary GH mRNA expression were also determined. Fasted channel catfish lost 5.6 and 15.6% body mass after 2 and 4 weeks of fasting, respectively. Plasma IGF-I concentrations were depressed ( P < 0.05) relative to fed controls following 2 and 4 weeks of fasting. Plasma GH concentrations were not different ( P > 0.05) in fasted fish after 2 weeks, but significantly increased ( P < 0.05) by 4 weeks of fasting. Hepatic IGF-I mRNA expression after 2 and 4 weeks of fasting was reduced ( P < 0.05) relative to fed controls. Pituitary GH mRNA expression was similar ( P > 0.05) between 2-week-fasted catfish and fed controls, but was increased ( P < 0.05) in 4-week-fasted catfish. The IGF-I TR-FIA was sensitive, accurate, and precise for all three species of fishes, and provided a low-cost, and non-radioisotopic method for quantifying plasma IGF-I levels in fed and fasted channel catfish.