Differential expression and activity of flavin-containing monooxygenases in euryhaline and stenohaline flatfishes indicates potential osmoregulatory role

Abstract

N,N-Dimethylaniline (DMA) N-oxidase activity indicative of flavin-containing monooxygenase (FMO) was examined in four tissues (liver, gill, muscle, and kidney) of the flounder ( Platichthys flesus). Gill microsomes had the highest levels of activity (456 ± 343 pmol/min/mg), while kidney (121± 109) and liver (67 ± 26) had levels just above detection. A single faint band of a 56 kD protein was observed in liver and gill microsomes following Western blot analyses with polyclonal antibodies to FMO 1. DMA N-oxidase activity in gill and liver directly correlated with the expression of the 56 kD protein recognized by polyclonal antibodies against FMO form 1. Likewise a mRNA band of approximately 2.5 kilobases was higher in gill than a 3.0 kb band in liver following hydridization with an FMO 1 cDNA probe. Gill and liver microsomal DMA N-oxidase from the euryhaline P. flesus was compared with that of the stenohaline turbot ( Scophthalmus maximus). DMA oxidase activity, FMO protein and mRNA were significantly greater in the gill of P. flesus, while S. maximus had higher levels of enzyme activity in the liver, but also significant levels in gill. Comparison of the enzymatic properties of the P. flesus gill and S. maximus liver enzymes indicated dramatic differences in K m between gill and liver, but were both inhibited by equimolar concentrations of trimethylamine (TMA). Gill microsomal activity in each species was unaffected by the mammalian FMO 2 substrate (competitive inhibitor) n-octylamine. Differential expression of FMO in tissues from stenohaline and euryhaline fish suggests a functional relationship between FMO and osmoregulation.