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Figures
- Fig. 1.
The effects of experimental treatments on circulating cortisol concentrations (ng ml–1) in gulf toadfish, Opsanus beta. In Series I (A), toadfish were injected with saline (N=8) or metyrapone (N=8). In Series II (B), toadfish were infused with saline (N=8), hydrocortisone hemisuccinate salt (cortisol; N=8), ammonia (N=8) or ammonia and cortisol combined (N=7). See Materials and methods for further details of treatment procedures. All values are expressed as means ± s.e.m. Data were analyzed by two-way RM-ANOVA with treatment group and sampling time as factors; P-values are indicated on the figure. Asterisks indicate a significant difference from the initial (0 h) sample within a given treatment group (Holm–Sidak post hoc test, P<0.05). Dashed vertical lines indicate the start of the treatment period.
- Fig. 2.
Measurements of (A,C) ammonia excretion (relative units) and (B,D) plasma ammonia levels (mmol l–1) in gulf toadfish, Opsanus beta, following various corticosteroid and ammonia treatments. In Series I (A,B), toadfish were treated with saline (N=8) or metyrapone (N=8). In Series II (C,D), toadfish were infused with saline (N=8), hydrocortisone hemisuccinate salt (cortisol; N=8), ammonia (N=8) or ammonia and cortisol combined (N=8). See Materials and methods for further details of treatment procedures. All ammonia excretion data are presented as the treatment value normalized to the initial 24 h control period. In A and C, numerical values presented under the data bars represent absolute values (μmol N kg–1) of ammonia excretion during the initial 24 h control period. All values are expressed as means ± s.e.m. Data were analyzed by two-way RM-ANOVA with treatment group and sampling time as factors; P-values are indicated on the figure. Asterisks indicate a significant difference between initial control and treatment values within a given treatment group (Holm–Sidak post hoc test, P<0.05). In B and D, the dashed vertical lines indicate the start of the treatment period.
- Fig. 3.
The effects of cortisol and/or ammonia manipulation on branchial and hepatic glutamine synthetase (GS) relative mRNA expression as measured by (A,C) real-time RT-PCR and (B,D) GS activity (mmol min–1 g–1) in gulf toadfish, Opsanus beta. In Series I (A,B), toadfish were injected with saline (N=8) or metyrapone (N=8). In Series II (C,D), toadfish were infused with saline (N=8), hydrocortisone hemisuccinate salt (cortisol; N=8), ammonia (N=8) or ammonia and cortisol combined (N=7). See Materials and methods for further details of treatment procedures. The mRNA transcript levels for each tissue were normalized against the control gene 18S and expressed relative to the saline-treated group for each series, which was set to a value of 1. Values are means ± s.e.m. For Series I, data were analyzed by Student's t-tests; see Results for P-values. No significant differences were detected for GS mRNA expression. For Series II, data were analyzed by one-way ANOVA; see Results for P-values. Statistical differences within a given tissue are indicated by different letters; different sets of letters were used between tissues (Holm–Sidak post hoc tests, P<0.05).
- Fig. 4.
Relative mRNA expression of Rhag, Rhbg, Rhcg1 and Rhcg2 in the gill of ureotelic gulf toadfish, Opsanus beta, as measured by real-time RT-PCR. In Series I (A), toadfish were injected with saline (N=8) or metyrapone (N=8). In Series II (B), toadfish were infused with saline (N=8), hydrocortisone hemisuccinate salt (cortisol; N=8), ammonia (N=8) or ammonia and cortisol combined (N=7). See Materials and methods for further details of treatment procedures. The mRNA levels for each gene were normalized against the control gene 18S and expressed relative to the saline-treated group for each series, which was set to a value of 1. Values are means ± s.e.m. For Series I, data were analyzed by Student's t-tests, and for Series II, data were analyzed by one-way ANOVA; see Results for P-values. Statistical differences for a given Rh isoform are indicated by different letters; different sets of letters were used between Rh isoforms (Series I, P<0.05; Series II, Holm–Sidak post hoc tests, P<0.05).
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