Department of Biological Science, National University of Singapore, Kent Ridge, Singapore 117543, Republic of Singapore.
J Exp Biol. 2009 Dec;212(Pt 23):3828-36. doi: 10.1242/jeb.034074.
This study aimed to examine whether the stenohaline freshwater stingray, Potamotrygon motoro, which lacks a functional ornithine-urea cycle, would up-regulate glutamine synthetase (GS) activity and protein abundance, and accumulate glutamine during a progressive transfer from freshwater to brackish (15 per thousand) water with daily feeding. Our results revealed that, similar to other freshwater teleosts, P. motoro performed hyperosmotic regulation, with very low urea concentrations in plasma and tissues, in freshwater. In 15 per thousand water, it was non-ureotelic and non-ureoosmotic, acting mainly as an osmoconformer with its plasma osmolality, [Na+] and [Cl-] comparable to those of the external medium. There were significant increases in the content of several free amino acids (FAAs), including glutamate, glutamine and glycine, in muscle and liver, but not in plasma, indicating that FAAs could contribute in part to cell volume regulation. Furthermore, exposure of P. motoro to 15 per thousand water led to up-regulation of GS activity and protein abundance in both liver and muscle. Thus, our results indicate for the first time that, despite the inability to synthesize urea and the lack of functional carbamoyl phosphate synthetase III (CPS III) which uses glutamine as a substrate, P. motoro retained the capacity to up-regulate the activity and protein expression of GS in response to salinity stress. Potamotrygon motoro was not nitrogen (N) limited when exposed to 15 per thousand water with feeding, and there were no significant changes in the amination and deamination activities of hepatic glutamate dehydrogenase. In contrast, P. motoro became N limited when exposed to 10 per thousand water with fasting and could not survive well in 15 per thousand water without food.
本研究旨在探讨是否缺乏功能性鸟氨酸-尿素循环的淡水尖吻鲷(Potamotrygon motoro)在从淡水向 15‰(千分之十五)盐水的渐进转移过程中,通过每日进食,会上调谷氨酰胺合成酶(GS)活性和蛋白丰度,并积累谷氨酰胺。我们的结果表明,与其他淡水硬骨鱼类一样,P. motoro 在淡水中进行高渗调节,血浆和组织中的尿素浓度非常低。在 15‰的盐水中,它是非尿氮排泄和非尿渗调节的,主要作为渗透调节者,其血浆渗透压、[Na+]和[Cl-]与外部介质相当。肌肉和肝脏中几种游离氨基酸(FAA)的含量(包括谷氨酸、谷氨酰胺和甘氨酸)显著增加,但血浆中没有增加,表明 FAA 可能部分有助于细胞体积调节。此外,P. motoro 暴露于 15‰的盐水中会导致肝脏和肌肉中 GS 活性和蛋白丰度的上调。因此,我们的结果首次表明,尽管 P. motoro 不能合成尿素,缺乏作为底物的功能性氨基甲酰磷酸合成酶 III(CPS III),但它仍具有在盐度胁迫下上调 GS 活性和蛋白表达的能力。当暴露于 15‰盐水并进食时,P. motoro 不会受到氮(N)限制,肝谷氨酸脱氢酶的氨化和脱氨活性也没有显著变化。相比之下,当暴露于 10‰盐水且禁食时,P. motoro 会受到 N 限制,在没有食物的情况下无法在 15‰盐水中很好地生存。