Increases in urea synthesis and the ornithine-urea cycle capacity in the giant African snail,Achatina fulica, during fasting or aestivation, or after the injection with ammonium chloride

Abstract

The objectives of this study are to determine whether a full complement of ornithine–urea cycle (OUC) enzymes is present in the hepatopancreas of the giant African snail Achatina fulica, and to investigate whether the rate of urea synthesis and the OUC capacity can be up‐regulated during 23 days of fasting or aestivation, or 24 hr post‐injection with NH₄Cl (10 μmol g⁻¹ snail) into the foot muscle. A. fulica is ureotelic and a full complement of OUC enzymes, including carbamoyl phosphate synthetase III (CPS III), was detected from its hepatopancreas. There were significant increases in the excretion of NH₄⁺, NH₃ and urea in fasting A. fulica. Fasting had no significant effect on the tissue ammonia contents, but led to a progressive accumulation of urea, which was associated with an 18‐fold increase in the rate of urea synthesis. Because fasting took place in the presence of water and because there was no change in water contents in the foot muscle and hepatopancreas, it can be concluded that the function of urea accumulation in fasting A. fulica was unrelated to water retention. Aestivation in arid conditions led to a non‐progressive accumulation of urea in A. fulica. During the first 4 days and the last 3 days of the 23‐day aestivation period, experimental snails exhibited significantly greater rates of urea synthesis compared with fasted snails. These increases were associated with significant increases in activities of various OUC enzymes, except CPS III, in the hepatopancreas. However, the overall urea accumulation in snails aestivated and snails fasted for 23 days were comparable. Therefore, the classical hypothesis that urea accumulation occurred to prevent water loss through evaporation during aestivation in terrestrial pulmonates may not be valid. Surprisingly, there were no accumulations of ammonia in the foot muscle and hepatopancreas of A. fulica 12 or 24 hr after NH₄Cl was injected into the foot muscle. In contrast, the urea content in the foot muscle of A. fulica increased 4.5‐ and 33‐fold at hour 12 and hour 24, respectively, and the respective increases in the hepatopancreas were 4.9‐ and 32‐fold. The exogenous ammonia injected into A. fulica was apparently detoxified completely to urea. The urea synthesis rate increased 148‐fold within the 24‐hr experimental period, which could be the greatest increase known among animals. Simultaneously, there were significant increases in activities of glutamine synthetase (2.5‐fold), CPS III (3.1‐fold), ornithine transcarbamoylase (2.3‐fold), argininosuccinate synthetase+lyase (13.6‐fold) and arginase (3.5‐fold) in the hepatopancreas 12 hr after the injection of NH₄Cl. Taken altogether, our results support the view that the primary function of urea synthesis through the OUC in A. fulica is to defend against ammonia toxicity, but suggest that urea may have more than an excretory role in terrestrial pulmonates capable of aestivation. J. Exp. Zool. 303A:1040;–1053, 2005.