The glass-eel stage of the life history of Anguillid eels (Anguilla spp.) makes the important physiological transition from seawater (SW) to freshwater (FW) osmoregulation. Central to successful adaptation to fresh water is the switch from active hypoosmoregulation (ion excretion) to active hyperosmoregulation (ion uptake) to counter passive fluxes imposed by concentration gradients. Anguilla anguilla (L.) glass eels, captured from the lower Minho River estuary and maintained in brackish water (BW; 24), were fully capable of adapting to FW following acute transfer. In a comparison between SW- (34) and FW-adapted glass eels, gill Na+/K+-ATPase and Na+:K+:2Cl cotransporter (NKCC) expression were both markedly reduced in the FW group. Branchial mitochondria-rich chloride cells (MRCs) were also significantly larger in SW-adapted glass eels. Apical CFTR Cl channel immunoreactivity associated with branchial MRCs was also only present in SW-adapted glass eels. The expression of these three ion-transport proteins is strongly associated with active ion excretion in SW teleost fishes. In FW-adapted glass eels, cortisol treatment increased branchial Na+/K+-ATPase expression but was without effect on NKCC expression. Glucocorticoid receptor blockade by RU-486 had no effect on Na+/K+-ATPase or NKCC expression.
To assess the role of noradrenaline (NA) as a possible regulator of brain energy metabolism in teleost fish, the impact of increased noradrenaline levels within the brain on several parameters of energy metabolism was assessed in rainbow trout brain. Accordingly, two different doses of noradrenaline, producing increases in brain NA levels comparable to those occurring in several physiological processes in nature, were selected. In a subsequent set of three different experiments, fish were intracerebroventricularly injected with 1 microL 100 g(-1) body weight of Cortland saline alone (control) or containing NA (5 nmol NA and 10 nmol NA); after 30 min, brain and plasma samples were taken to assess changes in parameters of energy metabolism due to NA treatment. The results obtained clearly show dose-dependent changes in NA-treated fish in several parameters, including decreased glycogen and ATP levels, increased lactate and pyruvate levels, decreased fructose 1,6-bisphosphatase activity, and increased pyruvate kinase and lactate dehydrogenase activities. Altogether, the present experiments show for the first time in a teleost fish evidence supporting that increased noradrenaline levels in the brain elicit metabolic changes in the brain (enhanced glycogenolysis and glycolysis), resulting in an increased energy demand. These metabolic changes may be related to those occurring under several physiological conditions in nature such as hypoxia, in which increased energy demand and increased noradrenaline levels occur in the brain simultaneously.
In the present study, glass eels Anguilla anguilla in the Minho River estuary (41Á5°N, 8Á5°W) decreased in size (standard length, L S and mass, M) from the beginning (autumn) to the end of the sampling season (summer). On the other hand elvers increased in L S and M from spring to summer and were significantly larger than glass eels in paired comparisons. Branchial Na þ /K þ -ATPase and vacuolar (V-type) proton ATPase (in vitro activities), two important ion transporting pumps, did not show significant seasonal changes in either glass eels or elvers although in glass eels Na þ /K þ -ATPase (activity) expression was significantly higher than in elvers. In a single month comparison Na þ /K þ -ATPase branchial mRNA expression was also higher in glass eels as was the protein level expression of both Na þ /K þ -ATPase and NKCC (Na þ :K þ :2Cl ÿ co-transporter). Immunofluorescence microscopy indicated apical CFTR Cl ÿ channel labelling in Na þ /K þ -ATPase positive chloride cell in glass eels which was absent in elvers. Whole body sodium concentration and percentage water did not show significant seasonal differences in either glass eels or elvers although there were significant differences between these two groups during some months.
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