ABSTRACT
Juvenile Senegalese sole Solea senegalensis were subjected for short periods to two different types of handling-related stress: air exposure stress and net handling stress. The S. senegalensis were sacrificed 2 and 24 h after the stress events and the levels of serotonin (5-HT), noradrenaline (NA), dopamine (DA) and their respective major metabolites, 5-hydroxyindoleacetic acid (5-HIAA), 3-methoxy-4-hydroxyphenylglycol (MHPG) and 3,4-dihydroxyphenylacetic acid (DOPAC), were measured in three brain regions (telencephalon, hypothalamus and optic tectum) and compared with those in control, non-stressed S. senegalensis. Neither type of stress caused any significant alteration of serotoninergic activity (5-HIAA:5-HT ratio) or NA levels. Dopaminergic activity (DOPAC:DA ratio) was lower in stressed fish in all of the brain regions studied. For both air exposure stress and net handling stress, DA levels were significantly higher (P < 0.05) than in the control S. senegalensis. In addition, the higher DA levels after net handling stress were always significantly higher (P < 0.05) than those observed after acute air exposure stress, except in the telencephalon after 24 h. The significantly lower DOPAC:DA ratio (P < 0.05) in all of the brain regions studied was only observed in response to net handling stress.
Subject(s)
Biogenic Monoamines/metabolism , Brain/metabolism , Flatfishes/metabolism , Neurotransmitter Agents/metabolism , Stress, Psychological/metabolism , 3,4-Dihydroxyphenylacetic Acid/metabolism , Animals , Dopamine/metabolism , Hydroxyindoleacetic Acid/metabolism , Hypothalamus/metabolism , Norepinephrine/metabolism , Serotonin/metabolismABSTRACT
The present study explored the potential role of brain catecholaminergic and serotoninergic systems as neuronal targets for the toxicological effects of acute ammonia exposure (0.28 mg l(-1) of un-ionized ammonia for 12 and 24 h) in juvenile sole (Solea senegalensis). In addition, plasma cortisol levels were measured. The results showed significant increases in their concentrations that were similar after 12 and 24 h of exposure. These data indicate that acute exposure (12 and 24 h) to ammonia initiates a typical stress response in the Senegalese sole, with stimulation of the hypothalamus-pituitary-interrenal axis. The concentrations of dopamine (DA), serotonin (5-hydroxytryptamine; 5-HT) and noradrenaline (NA), and their metabolites, 3, 4-dihydroxyphenylacetic acid (DOPAC) and 5-hydroxy-3-indoleacetic acid (5HIAA), were measured in the hypothalamus, telencephalon and optic tectum. The main changes induced by acute exposure to ammonia were decreases in the concentrations of 5-HT and DA, which were significant in most of the brain regions studied. The ratios of 5-HIAA/5-HT and DOPAC/DA increased in all regions and at all times studied, although in the case of the DOPAC/DA ratio, the increases were only significant in the hypothalamus (24 h exposure) and in the optic tectum (12 and 24 h exposure). These changes indicated that exposure to ammonia elicited time-dependent increases in serotoninergic and dopaminergic activity in the hypothalamus, telencephalon and optic tectum.
Subject(s)
Ammonia/toxicity , Biogenic Monoamines/metabolism , Brain/drug effects , Flatfishes/physiology , Neurotransmitter Agents/metabolism , Water Pollutants, Chemical/toxicity , 3,4-Dihydroxyphenylacetic Acid/metabolism , Ammonium Chloride/toxicity , Animals , Brain/metabolism , Brain Chemistry , Dopamine/metabolism , Hydrocortisone/blood , Hydroxyindoleacetic Acid/metabolism , Hypothalamo-Hypophyseal System/drug effects , Hypothalamo-Hypophyseal System/metabolism , Kidney/drug effects , Kidney/metabolism , Life Cycle Stages/drug effects , Life Cycle Stages/physiology , Norepinephrine/metabolism , Serotonin/metabolismABSTRACT
An enzymoimmunoassay (EIA) kit for plasma melatonin (MLT) measurements was employed in tench (Tinca tinca) and in turbot (Scophtalmus maximus). Tench and turbot plasma samples were purified with a C18 reversed phase extraction columns because this kit is designed for human serum measurements. The lowest detection limit of the technique was 11.48 pg/well with a sensitivity at 50% binding of 100 pg/well. Intra-assay and inter-assay CV (%) were always less than 5% (n=8), and 9% (n=6) in tench plasma samples, and less than 5% (n=8) and 13% (n=5) in turbot plasma samples, respectively. Correlation coefficients between EIA and RIA measurements in tench and turbot plasma samples were 0.93 and 0.89 (p<0.001) respectively. Diurnal and nocturnal plasma melatonin mean levels were 14.7+/-2.1 pg/ml and 87.4+/-11 pg/ml in tench (n=15), and 3.5+/-0.4 pg/ml and 28.1+/-2.1 pg/ml in turbot (n=15). These species showed a melatonin circadian rhythm as in other animals studied. The results suggest that the commercial kit used in this experiment could be a suitable and alternative method to RIA for plasma MLT determinations in tench and turbot although it is necessary to increase volumes (1ml) and concentrate daytime samples.
