Acute and chronic temperature dependence of Na+/H+ exchange activity of Pimephales promelas gills

Abstract

Na⁺/H⁺ exchangers (NHE) mediate at least part of Na⁺ entry into gill epithelia via Na⁺/NH₄⁺ exchange. For homeostasis, Na⁺ entry into and exit via Na⁺/K⁺ ATPase from gill epithelia must balance. Na⁺/K⁺ ATPase activity is reduced in cold- compared to warm-acclimated freshwater temperate fish. We hypothesized gill NHE activity is greater in warm- than cold-acclimated fish when measured at acclimation temperatures, and NHE activity displays a temperature dependence similar to Na⁺/K⁺ ATPase. Since NHE mRNA expression does not differ, we measured the Na⁺-dependence of pH-induced Na⁺ fluxes in gill vesicles from warm- and cold-acclimated fathead minnows at 20^o and 7 °C, and calculated maximum transport rates (V_max) and Na⁺ K_1/2s. We also measured NH₄⁺-induced Na⁺ fluxes and Na⁺-induced H⁺ fluxes. In vesicles from warm-acclimated fish, NHE V_maxs were 278 ± 33 and 149 ± 23 arbitrary unit/s (au/s) and Na⁺ K_1/2s were 12 ± 4 and 6 ± 4 mmol/l when assayed at 20^o and 7 °C (p < 0.004), respectively. In vesicles from cold-acclimated fish, V_maxs were 288 ± 35 and 141 ± 13 au/s and Na⁺ K_1/2s 17 ± 5 and 7 ± 2 mmol/l when assayed at 20^o and 7 °C (p < 0.002), respectively. Na⁺-induced H⁺ fluxes were 98 ± 8 and 104 ± 26 au/s in warm- and cold-acclimated fish assayed at 20 °C, respectively. Na⁺/NH₄⁺ exchange was 120 ± 11 and 158 ± 13 au/s in warm- and cold-acclimated fish, respectively. Conclusions: Gill NHE activity was greater in warm- than cold-acclimated fish assayed at acclimation temperatures. The temperature dependence of NHE activity was similar in both groups, but differed from that reported for Na⁺/K⁺ ATPase suggesting complex mechanisms to maintain Na⁺ homeostasis.

Keywords: Ammonium; Na absorption; Na transport; Teleost.