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In vivo characterisation of intestinal zinc uptake in freshwater rainbow trout

Chris N. Glover^1,* and Christer Hogstrand^1,2

¹ Division of Life Sciences, King’s College, London, London SE1 9NN, UK and
² T. H. Morgan School of Biological Sciences, University of Kentucky, Lexington, Kentucky 40506–0225, USA

View larger version (12K): [in a new window]   Fig. 1. Profile of 65Zn(II) activity obtained from the efferent cannula following perfusion of the intestine with Zn(II) (50 µmol l–1). Perfusion rates averaged 9.0±0.15 ml h–1. The vertical line indicates when the solution was switched to 65Zn(II)-free medium. Values are means ± S.E.M. (N=10, Zn(II) perfusion; N=4, Zn(II)-free perfusion).

View larger version (20K): [in a new window]   Fig. 2. The influence of perfused Zn(II) concentration upon epithelial mass, retained Zn(II) fraction and absorbed Zn(II) fraction. The value for epithelial mass (g dry mass) resulting from perfusion of Zn(II)-free solutions (filled bar) is the mean ± S.E.M. of 4 experiments; epithelial mass of Zn(II) perfused solutions (open bars), retained Zn(II) fraction (open circles), and absorbed Zn(II) fraction (filled circles) are means ± S.E.M. of 7–11 experiments. Control and Zn(II)-exposed intestines were perfused for 3 h, and retained and absorbed fractions are expressed as percentage of total Zn(II) perfused over the 3 h period. Epithelial mass is the mass of material scraped from the intestinal mucosal surface and includes both epithelial cells and mucus. Retained Zn(II) fraction is the proportion of perfused Zn(II) retained in the animal, while absorbed Zn(II) fraction represents the proportion of Zn(II) retained in post-epithelial tissues (intestine, blood, body).

View larger version (12K): [in a new window]   Fig. 3. Net influx of Zn(II) (nmol g–1 h–1) from the perfused medium into the tissues, measured as disappearance of Zn(II) from perfusate. Values are means ± S.E.M. (N=7–11). Net influx was calculated over the final 30 min of a 3 h perfusion.

View larger version (13K): [in a new window]   Fig. 4. Epithelial Zn(II) accumulation rate (nmol g dry mass–1 h–1) following a 3 h intestinal perfusion of the intestine with Zn(II) solutions of varying concentration. Values are means ± S.E.M. (N=7–11). Epithelium represents tissue scraped from intestinal mucosal surface and includes epithelial cells and surface mucus.

View larger version (12K): [in a new window]   Fig. 5. Subepithelial Zn(II) accumulation rate (nmol g–1 wet mass h–1) following a 3 h perfusion of the intestine with Zn(II) solutions of varying concentration. Values are means ± S.E.M. (N=7–11). Subepithelium represents intestinal tissue remaining following mucosal scraping.

View larger version (13K): [in a new window]   Fig. 6. Body Zn(II) accumulation rate (nmol g–1 wet mass h–1) following a 3 h perfusion of the intestine with Zn(II) of varying concentration. Values are means ± S.E.M. (N=7–11). Body represents the tissues remaining following intestinal excision.

View larger version (18K): [in a new window]   Fig. 7. Relationship between subepithelial (intestinal tissue following mucosal scraping) and post-intestinal (pooled blood and body) Zn(II) accumulation at different concentrations of perfused Zn(II). Values are means ± S.E.M. (N=7–11).

View larger version (19K): [in a new window]   Fig. 8. Effect of post-perfusion flushing of excised intestine on epithelial (A) and subepithelial (B) Zn(II) accumulation rate (nmol g–1 h–1). Values are means ± S.E.M. (N=3–4). Significant differences * between perfused Zn(II) concentrations; between control and flushed treatments; between EGTA-free and EGTA solutions, where indicated, are at P<0.05.