TY - JOUR
T1 - 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
AU - Zimmer, Alex M.
AU - Nawata, C. Michele
AU - Wood, Chris M.
N1 - Funding Information:
Acknowledgments This study was funded by an NSERC Discovery Grant to CMW, who is also supported by the Canada Research Chair Program. The experiments were approved by the institutional animal care committee at McMaster University.
PY - 2010
Y1 - 2010
N2 - Recently, a "Na+/NH4+ 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 CO2 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 NH4HCO3 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.
AB - Recently, a "Na+/NH4+ 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 CO2 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 NH4HCO3 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.
KW - Ammonia excretion
KW - Feeding
KW - High environmental ammonia (HEA)
KW - Rainbow trout
KW - Rhesus glycoproteins
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U2 - 10.1007/s00360-010-0488-4
DO - 10.1007/s00360-010-0488-4
M3 - Article
C2 - 20563818
AN - SCOPUS:77958470477
VL - 180
SP - 1191
EP - 1204
JO - Journal of Comparative Physiology B: Biochemical, Systemic, and Environmental Physiology
JF - Journal of Comparative Physiology B: Biochemical, Systemic, and Environmental Physiology
SN - 0174-1578
IS - 8
ER -