title: An in vitro analysis of intestinal ammonia handling in fasted and fed freshwater rainbow trout (Oncorhynchus mykiss).
abstract: Ammonia transport and metabolism were investigated in the intestinal tract of freshwater rainbow trout which had been either fasted for 7 days, or fasted then fed a satiating meal of commercial trout pellets. In vivo, total ammonia concentrations (T amm) in the chyme were approximately 1 mmol L(-1) across the entire intestine at 24 h after the meal. Highest chyme pH and P NH3 values occurred in the posterior intestine. In vitro gut sac experiments examined ammonia handling with mucosal (Jmamm) and serosal (Js amm) fluxes under conditions of fasting and feeding, with either background (control ≤ 0.013 mmol L(-1)) or high luminal ammonia concentrations (HLA = 1 mmol L(-1)), the latter mimicking those seen in chyme in vivo. Feeding status (fasted or fed) appeared to influence ammonia handling by each individual section. The anterior intestine exhibited the greatest Jm amm and Js amm values under fasted control conditions, but these differences tended to disappear under typical post-feeding conditions when total endogenous ammonia production (Jt amm = Js amm - Jm amm, signs considered) was greatly elevated in all intestinal sections. Under fasted conditions, glutamate dehydrogenase (GDH) and glutaminase (GLN) activities were equal across all sections, but the ammonia-trapping enzyme glutamine synthetase (GS) exhibited highest activity in the posterior intestine, in contradiction to previous literature. Feeding clearly stimulated the total rate of endogenous ammonia production (Jt amm), even in the absence of a high luminal ammonia load. This was accompanied by an increase in GDH activity of the anterior intestine, which was also the site of the largest Jt amm. In all sections, during HLA exposure, either alone or in combination with feeding, there were much larger increases in endogenous Jt amm, most of which was effluxed to the serosal solution. This is interpreted as a response to avoid potential cytotoxicity due to overburdened detoxification mechanisms in the face of elevated mucosal ammonia. Thus T amm of the intestinal tissue remained relatively constant regardless of feeding status and exposure to HLA. Ammonia production by the gut may explain up to 18 % of whole-body ammonia excretion in vivo under fasting conditions, and 47 % after feeding, of which more than half originates from endogenous production rather than from absorption from the lumen.
authors.affiliation: Department of Biology, McMaster University, Hamilton, ON, L8S 4K1, Canada, rubinojg@gmail.com.
authors.forename: Julian G
authors.initials: JG
authors.lastname: Rubino
authors.forename: Alex M
authors.initials: AM
authors.lastname: Zimmer
authors.forename: Chris M
authors.initials: CM
authors.lastname: Wood
mesh_terms: D000641:Ammonia / Q000378:metabolism / Q000494:pharmacology*; D000818:Animals; D005215:Fasting / Q000378:metabolism*; D029941:Fish Proteins / Q000378:metabolism; D005969:Glutamate Dehydrogenase / Q000378:metabolism; D005974:Glutamate-Ammonia Ligase / Q000378:metabolism; D005972:Glutaminase / Q000378:metabolism; D066298:In Vitro Techniques; D007413:Intestinal Mucosa / Q000378:metabolism*; D017686:Oncorhynchus mykiss / Q000378:metabolism*
chemical_list: D029941:Fish Proteins; D000641:Ammonia; D005969:Glutamate Dehydrogenase; D005972:Glutaminase; D005974:Glutamate-Ammonia Ligase
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