SUMMARYThe swamp eel, Monopterus albus, is an air-breathing teleost which typically lives in freshwater but can also be found in estuaries, where it has to deal with ambient salinity fluctuations. Unlike other teleosts, its gills are highly degenerate. Hence, it may have uncommon osmoregulatory adaptations, but no information is available on its osmoregulatory capacity and mechanisms at present. In this study M. albus was exposed to a 5 day progressive increase in salinity from freshwater (1‰) to brackish water ( . Consequently, fish exposed to 25‰ water had to undergo cell volume regulation through accumulation of organic osmolytes and inorganic ions. Increases in tissue free amino acid content were apparently the result of increased protein degradation, decreased amino acid catabolism, and increased synthesis of certain non-essential amino acids. Here we report for the first time that glutamine is the major organic osmolyte in M. albus. Glutamine content increased to a phenomenal level of >12 μmol g -1 and >30 μmol g -1 in the muscle and liver, respectively, of fish exposed to 25‰ water. There were significant increases in glutamine synthetase (GS) activity in muscle and liver of these fish. In addition, exposure to 25‰ water for 4 days led to significant increases in GS protein abundance in both muscle and liver, indicating that increases in the expression of GS mRNA could have occurred.

The objective of this study was to elucidate the role of the intestine from juveniles of the marble goby, Oxyeleotris marmorata, during seawater (SW) exposure. It has been reported elsewhere that SW-exposed juvenile O. marmorata exhibits hypoosmotic and hypoionic regulation, with the induction of branchial Na(+)/K(+)-ATPase (NKA), Na(+):K(+):2Cl(-) cotransporter (NKCC), and cystic fibrosis transmembrane receptor-like chloride channels. Here, we report that SW exposure also led to significant increases in the activity and protein abundance of NKA in, and probably an increase in Na(+) uptake through, its intestine. Additionally, there was an increase in apical NKCC immunoreactivity in the intestinal epithelium, indicating that there could be increased Cl(-) uptake through the intestine. These results suggest that absorption of ions, and hence water, from the intestinal lumen could be an essential part of the osmoregulatory process in juvenile O. marmorata during exposure to SW. Furthermore, there were significant increases in the glutamate content, and the aminating activity and protein abundance of glutamate dehydrogenase (GDH) in the intestine of fish exposed to SW. Since the intestinal glutamine synthetase activity and protein abundance decreased significantly, and the intestinal glutamine content remained unchanged, in the SW-exposed fish, excess glutamate formed via increased GDH activity in the intestine could be channeled to other organs to facilitate the increased synthesis of amino acids. Taken together, our results indicate for the first time that, besides absorbing ions and water during SW exposure, the intestine of juvenile O. marmorata also participated in altered nitrogen metabolism in response to salinity changes.

The objectives of this study were to examine how the swamp eel, Monopterus albus, defended against acute ammonia toxicity derived from the intraperitoneal injection with a sublethal dose (10 micromol g(-1) fish) of ammonium acetate (CH(3)COONH(4)) followed by 24 hr of emersion, and to elucidate the mechanisms of acute ammonia toxicity with respect to glutamine accumulation in the brain using L-methionine S-sulfoximine [MSO; a glutamine synthetase inhibitor]. When confronted with a sublethal dose of CH(3)COONH(4) followed by emersion, only a small fraction of the exogenous ammonia was excreted, and ammonia contents in various organs, especially the brain, increased transiently to high levels. Increased glutamine synthesis and decreased amino acid catabolism in and outside the brain were involved in the defence against acute ammonia toxicity. When injected with a lethal dose (16 micromol g(-1) fish) of CH(3)COONH(4) followed by emersion, ammonia (approximately 30 micromol g(-1) tissue), but not glutamine ( approximately 5 micromol g(-1) tissue), accumulated to extraordinarily high levels in the brain of succumbed fish. Hence, glutamine accumulation in the brain might not be the major mechanism of acute ammonia toxicity in M. albus. MSO (100 microg g(-1) fish) had a partial protective effect in fish injected with a lethal dose of CH(3)COONH(4). However, this effect was unrelated to the suppression of glutamine synthesis and accumulation in the brain. Instead, MSO suppressed the rate of ammonia buildup in the brain, possibly through its effects on glutamate dehydrogenase therein.

The swamp eel, Monopterus albus, is an obligatory air-breathing teleost which can undergo long period of emersion, has high environmental and tissue ammonia tolerance, and can survive in brackish water. We obtained a cDNA sequence of glutamate dehydrogenase (gdh), which consisted of a 133-bp 5′ UTR, a complete coding sequence region spanning 1629 bp and a 3′ UTR of approximately 717 bp, from the liver, intestine, and brain of M. albus. The translated Gdh amino acid sequence had 542 residues, and it formed a monophyletic clade with Bostrychus sinensis Gdh1a, Tetraodon nigroviridis Gdh1a, Chaenocephalus aceratus Gdh1a, Salmo salar Gdh1a1 and Gdh1a2, and O. mykiss Gdh1a. One day of exposure to terrestrial conditions or 75 mmol l−1 NH4Cl, but not to water at salinity 20, resulted in a significant increase in mRNA expression of gdh1a and Gdh amination activity in the liver of M. albus. However, exposure to brackish water, but not to terrestrial conditions or 75 mmol l−1 NH4Cl, led to a significant increase in the mRNA expression of gdh1a and Gdh amination activity in the intestine. By contrast, all the three experimental conditions had no significant effects on the mRNA expression of gdh1a in the brain of M. albus, despite a significant decrease in the Gdh amination activity in the brain of fish exposed to 75 mmol l−1 NH4Cl for 6 days. Our results indicate for the first time that the mRNA expression of gdh1a was differentially up-regulated in the liver and intestine of M. albus in response to ammonia toxicity and salinity stress, respectively. The increases in mRNA expression of gdh1a and Gdh amination activity would probably lead to an increase in glutamate production in support of increased glutamine synthesis for the purpose of ammonia detoxification or cell volume regulation under these two different environmental conditions.