Metabolic and neuroendocrine effects on diurnal urea excretion in the mangrove killifish Rivulus marmoratus

In mangrove killifish Rivulus marmoratus, urea excretion (J(urea)) follows a distinct diurnal pattern with the highest rates between 12:00 h and 18:00 h. We investigated the regulating mechanisms that underlie temporal rhythms in J(urea) in R. marmoratus. We hypothesized that the daily pattern of J(urea) in R. marmoratus is (1) due to diurnal changes in urea synthesis rates and ultimately metabolic rate and/or (2) controlled by neuroendocrine messengers. Oxygen consumption and whole body urea content in R. marmoratus demonstrated a clear diurnal pattern with maximum rates for both parameters occurring at 12:00 h. A strong synchrony between diurnal patterns of oxygen consumption, whole body urea content and J(urea) implicated metabolic regulation of the diurnal J(urea) pattern. Ketanserin, a 5-HT(2) receptor antagonist, and RU-486, a cortisol receptor antagonist, were used to test the second hypothesis. Increasing antagonist concentrations of either ketanserin or RU-486 resulted in dose-dependent decreases in J(urea). Application of a single dose of either antagonist significantly decreases J(urea) for up to 12 and 6 h for ketanserin and RU-48, respectively. Repeated exposure to doses of either ketanserin or RU-486 did not abolish the diurnal pattern in J(urea); however, there was a significant decrease in the amplitude of the rates. Taken together, these findings indicate that the diurnal pattern of J(urea) in R. marmoratus are regulated by both metabolic and neuroendocrine factors. We propose that cortisol and 5-HT influence the absolute rate of urea excretion by altering the permeability of the gill membrane to urea and/or the rate of urea synthesis.