title: Physiological and molecular analysis of the interactive effects of feeding and high environmental ammonia on branchial ammonia excretion and Na+ uptake in freshwater rainbow trout.
abstract: Recently, a "Na(+)/NH(4)(+) exchange complex" model has been proposed for ammonia excretion in freshwater fish. The model suggests that ammonia transport occurs via Rhesus (Rh) glycoproteins and is facilitated by gill boundary layer acidification attributable to the hydration of CO(2) and H(+) efflux by Na(+)/H(+) exchanger (NHE-2) and H(+)-ATPase. The latter two mechanisms of boundary layer acidification would occur in conjunction with Na(+) influx (through a Na(+) channel energized by H(+)-ATPase and directly via NHE-2). Here, we show that natural ammonia loading via feeding increases branchial mRNA expression of Rh genes, NHE-2, and H(+)-ATPase, as well as H(+)-ATPase activity in juvenile trout, similar to previous findings with ammonium salt infusions and high environmental ammonia (HEA) exposure. The associated increase in ammonia excretion occurs in conjunction with a fourfold increase in Na(+) influx after a meal. When exposed to HEA (1.5 mmol/l NH(4)HCO(3) at pH 8.0), both unfed and fed trout showed differential increases in mRNA expression of Rhcg2, NHE-2, and H(+)-ATPase, but H(+)-ATPase activity remained at control levels. Unfed fish exposed to HEA displayed a characteristic reversal of ammonia excretion, initially uptaking ammonia, whereas fed fish (4 h after the meal) did not show this reversal, being able to immediately excrete ammonia against the gradient imposed by HEA. Exposure to HEA also led to a depression of Na(+) influx, demonstrating that ammonia excretion can be uncoupled from Na(+) influx. We suggest that the efflux of H(+), rather than Na(+) influx itself, is critical to the facilitation of ammonia excretion.
authors.affiliation: Department of Biology, McMaster University, Hamilton, ON, L8S 4K1, Canada. zimmea@mcmaster.ca
authors.forename: Alex M
authors.initials: AM
authors.lastname: Zimmer
authors.forename: C Michele
authors.initials: CM
authors.lastname: Nawata
authors.forename: Chris M
authors.initials: CM
authors.lastname: Wood
mesh_terms: D000641:Ammonia / Q000008:administration & dosage / Q000378:metabolism / Q000494:pharmacology*; D000818:Animals; D004435:Eating / Q000502:physiology*; D029941:Fish Proteins / Q000378:metabolism; D005880:Gills / Q000201:enzymology / Q000502:physiology; D006023:Glycoproteins / Q000378:metabolism; D017686:Oncorhynchus mykiss / Q000378:metabolism; D006180:Proton-Translocating ATPases / Q000378:metabolism; D012964:Sodium / Q000378:metabolism*; D017923:Sodium-Hydrogen Exchangers / Q000378:metabolism
chemical_list: D029941:Fish Proteins; D006023:Glycoproteins; D017923:Sodium-Hydrogen Exchangers; D000641:Ammonia; D012964:Sodium; D006180:Proton-Translocating ATPases