Physical characterization of high-affinity gastrointestinal Cu transport in vitro in freshwater rainbow trout Oncorhynchus mykiss

This study investigated the transport of copper (Cu) in the gut of trout. Examination of the spatial distribution of Cu along the digestive tract and a physical characterization of the uptake process was carried out using an in vitro gut sac technique and 64Cu as a tracer. Unidirectional Cu uptake was highest in the anterior intestine followed in decreasing order by the posterior intestine, mid intestine and the stomach. Cu uptake was resistant to hypoxia and appeared to be fueled equally well by Cu(II) or Cu (I) at Cu concentrations typically found in the fluid phase of the chyme in vivo in the trout intestine. Transport demonstrated saturation kinetics (e.g. K m = 31.6 μM, J max = 17 pmol cm−2 h−1, in mid intestine) at low Cu levels representative of those measured in the chyme in vivo, with a diffusive component at higher Cu concentrations. Q 10 analysis indicated Cu uptake is via diffusion across the apical membrane and biologically mediated across the basolateral membranes of enterocytes. The presence of l-histidine but not d-histidine stimulated both Cu and Na uptake suggesting a common pathway for the transport of Cu/Na with l-histidine

In vitro examination of interaction between copper and zinc uptake via the gastrointestinal tract of rainbow trout (Oncorhynchus mykiss).

An in vitro gut sac technique was used to investigate whether reciprocal inhibitory effects occurred between Cu and Zn uptake in the gastrointestinal tract of the rainbow trout and, if so, whether there was regional variation among the stomach, anterior intestine, mid intestine, and posterior intestine in the phenomena. Metal accumulation in surface mucus and in the mucosal epithelium and transport into the blood space were assayed using radiolabeled Cu or Zn at environmentally realistic concentrations of 50 μmol L−1 in the luminal saline, with 10-fold higher levels of the other metal (nonradioactive) as a potential inhibitor. Zn transport rates were generally higher than Cu transport rates in all compartments except the stomach, where they were lower. High [Zn] reduced Cu transport into the blood space in the mid and posterior intestines by 67% and 33%, respectively, whereas high [Cu] reciprocally reduced Zn transport into the blood space in these same sections by 54% and 78%. No inhibitions occurred in either the anterior intestine or the stomach. In these segments, elevated concentrations of the other metal stimulated Cu and Zn transport into the blood space and/or the mucosal epithelium by 50–100%, possibly by displacement from intracellular binding sites, thereby raising local concentrations at other transport sites. None of the treatments affected metal accumulation in surface mucus. The results indicate that one or more shared high-affinity pathways (possibly DMT1) occur in the mid and posterior intestine, which transport both Cu and Zn. These pathways appear to be absent from the stomach and anterior intestine, where other transport mechanisms may occur.NSER

Gastrointestinal assimilation of Cu during digestion of a single meal in the freshwater rainbow trout ( Oncorhynchus mykiss)

Gastrointestinal processing and assimilation of Cu in vivo was investigated by sequential chyme analysis over a 72 h period following ingestion of a single satiation meal (3% body weight) of commercial trout food (Cu content = 0.42 μmol g − 1 ) by adult rainbow trout. Leaded glass ballotini beads incorporated into the food and detected by X-ray radiography were employed as an inert marker in order to quantify net Cu absorption or secretion in various parts of the tract. Cu concentrations in the supernatant (fluid phase) fell from about 0.06 μmol mL − 1 (63 μM) in the stomach at 2 h to about 0.003 μmol mL − 1 (3 μM) in the posterior intestine at 72 h. Cu concentrations in the solid phase were 10 to 30-fold higher than in the fluid phase, and increased about 4-fold from the stomach at 2 h to the posterior intestine at 72 h. By reference to the inert marker, overall net Cu absorption from the ingested food by 72 h was about 50%. The mid-intestine, and posterior intestine emerged as important sites of net Cu and water absorption and a potential role for the stomach in this process was also indicated. The anterior intestine was a site of large net Cu addition to the chyme, probably due to large net additions of Cu-containing fluids in the form of bile and other secretions in this segment. The results provide valuable information about sites of Cu absorption and realistic concentrations of Cu in chyme fluid for future in vitro mechanistic studies on Cu transport in the trout gastrointestinal tract