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Figures
- Fig. 1.
Aquatic hypoxia and air exposure causes changes in the blood of Kryptolebias marmoratus. (A) Hb protein concentration, (B) hematocrit and (C) red blood cell counts. Kryptolebias marmoratus were acclimated for 7 days to one of four treatment groups: normoxic water, normoxic air, hypoxic water or hypoxic air. Hypoxia in water (PO2=3.59±0.06kPa) and air (PO2=13.57±0.06kPa) was achieved by mixing N2 gas with atmospheric air. Different lower case letters indicate significant overall differences between normoxia and hypoxia (two-way ANOVA, P<0.05). Different upper case letters indicate significant differences between the four treatment groups if a significant interaction between oxygen availability and respiratory medium was found (two-way ANOVA, P<0.05). Asterisks denote a significant overall difference between fish in water versus air (two-way ANOVA, P<0.05). Sample sizes are given in parentheses. Error bars represent s.e.m.
- Fig. 2.
Air exposure increases the affinity of hemoglobin for O2 in Kryptolebias marmoratus. Fish were acclimated for 7 days to one of four treatment groups: normoxic water, normoxic air, hypoxic water or hypoxic air. Hypoxia in water (PO2=3.59±0.06kPa) and air (PO2=13.57±0.06kPa) was achieved by mixing N2 gas with atmospheric air. Different upper case letters indicate significant overall differences in P50 (i.e. across all concentrations of CO2, not within one PCO2) between the four treatment groups when a significant interaction between oxygen availability and respiratory medium was found (three-way ANOVA, P<0.05). P50 is presented in Torr; 1Torr=133Pa. There was a significant overall difference in P50 between each concentration of CO2 used (three-way ANOVA, P<0.05), but there was no significant interaction between CO2 and oxygen availability (P=0.71) or between CO2 and respiratory medium (P=0.82). Sample sizes are given in parentheses. Error bars represent s.e.m.
- Fig. 3.
Air exposure induces angiogenesis in the cutaneous epithelium and bucco-opercular chamber of Kryptolebias marmoratus. Representative composite images are presented of immunohistochemical staining for the angiogenesis marker CD31 (green) in a sagittal section through the head of Kryptolebias marmoratus acclimated to normoxic water (A) or air (B) for 7 days. Composite images were created from low-magnification (×4) fluorescent and differential interference contrast photomicrographs stitched together and overlaid using Adobe Photoshop. (C) Quantification of CD31 fluorescence (arbitrary units; a.u.) in fish from one of four treatment groups: normoxic water, hypoxic water, normoxic air or hypoxic air. Different letters indicate significant overall differences between fish in water versus air (three-way ANOVA, P<0.05). Asterisks denote a significant overall difference (three-way ANOVA, P<0.05) between the ventral epithelium and all other body regions. Error bars represent s.e.m.
- Fig. 4.
Aerial ventilation and body movements in Kryptolebias marmoratus exposed to air. Frequency of aerial ventilation (A) and number of body movements (B) was measured over 7 days. Fish (N=8) were acclimated to air for 1 h, 1 day or 7 days. (C) Cumulative frequency of aerial ventilations in relation to whole body movements in 1 h recording periods after 1 h, 1 day and 7 days of air exposure. Significant differences (one-way ANOVA, P<0.05) between time points are indicated by different letters. Sample sizes are given in parentheses. Error bars represent s.e.m.
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