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First published online October 7, 2008
Journal of Experimental Biology 211, 3237-3248 (2008)
Published by The Company of Biologists 2008
doi: 10.1242/jeb.019257
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Differential recovery from exercise and hypoxia exposure measured using 31P- and 1H-NMR in white muscle of the common carp Cyprinus carpio

Troy M. Hallman, Anibal C. Rojas-Vargas, David R. Jones and Jeffrey G. Richards*

Department of Zoology, The University of British Columbia, 6270 University Boulevard, Vancouver, BC, Canada V6T 1Z4


Figure 1
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  		Fig. 1. Representative series of spectra obtained from white muscle of carp acclimated to 15°C and exposed to hypoxia followed by recovery (A) or during recovery from exhaustive exercise (B). The first spectra in panel A represent metabolite profiles in our control group (rest/normoxia) and the first spectra in panel B represents exhausted fish. PCr, phosphocreatine; Pi, intracellular phosphate.

 

Figure 2
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  		Fig. 2. White muscle PCr (A) and pHi (B) following exercise (white bars) and exposure to hypoxia (striped bars) in carp acclimated to 15 or 25°C. Phosphocreatine levels are normalized to the sum of PCr and Pi, and expressed relative to normoxic/resting controls. NMR data were collected at the fishes' acclimation temperature. Each bar represents mean + s.e.m. Significant differences (P<0.05) between exercise and hypoxia at a given temperature are indicated by + (two-way ANOVA). From left to right in each panel, N=7, 7, 10 and 6.

 

Figure 3
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  		Fig. 3. White muscle PCr (A) and pHi (B) in carp exposed to normoxia and hypoxia for up to 3 h at 15°C (squares) and 25°C (circles). Phosphocreatine levels are normalized to the sum of PCr and Pi, and expressed relative to normoxic controls. NMR data were collected at the fishes' acclimation temperature and for each time point an average consisting of 256 spectra gathered over 5 min is shown. The black horizontal bar immediately above the x-axis indicates the period of hypoxia exposure (PO2=20 Torr). Each point represents mean ± s.e.m. N=7–10 for each point.

 

Figure 4
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  		Fig. 4. Recovery of PCr following exercise (A) and hypoxia exposure (B) at 15°C (squares) and 25°C (circles). Phosphocreatine levels are normalized to the sum of PCr and Pi and expressed relative to normoxic/resting controls. NMR data were collected at the fishes' acclimation temperature and for each time point an average consisting of 256 spectra gathered over 5 min is shown. Time zero represents the point at which exercise stopped (A) or when fish were returned to normoxic water (B). In A, transfer of the fish from the exercise respirometer to the NMR took ~10 min, therefore the recovery trace does not start at zero. Each point represents mean ± s.e.m. N=7–10 for each data point.

 

Figure 5
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  		Fig. 5. Recovery of pHi following exercise (A) and hypoxia exposure (B) at 15°C (squares) and 25°C (circles). Each point represents mean ± s.e.m. N=7–10 for each data point. See Fig. 4 legend for more detail.

 

Figure 6
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  		Fig. 6. Recovery of ATP following exercise (A) and hypoxia exposure (B) at 15°C (squares) and 25°C (circles). ATP levels are expressed relative to normoxic/resting controls. Each point represents mean ± s.e.m. N=7–10 for each data point. See Fig. 4 legend for more detail.

 

Figure 7
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  		Fig. 7. Recovery of cytosolic free ADP following exercise (A) and hypoxia exposure (B) at 15°C (squares) and 25°C (circles). ADPfree levels are in units of µmol l–1 intracellular water (see Materials and methods for more detail). Each point represents mean ± s.e.m. N=7–10 for each data point. See Fig. 4 legend for more detail.

 

Figure 8
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  		Fig. 8. Recovery of [ATP]/[ADPfree] following exercise (A) and hypoxia exposure (B) at 15°C (squares) and 25°C (circles). Each point represents mean ± s.e.m. N=7–10 for each data point. See Fig. 4 legend for more detail.

 

Figure 9
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  		Fig. 9. Recovery of Gibbs free energy of ATP hydolysis ({Delta}fG'; kJ mol–1) following exercise (A) and hypoxia exposure (B) at 15°C (squares) and 25°C (circles). Each point represents mean ± s.e.m. N=7–10 for each data point. See Fig. 4 legend for more detail.

 

Figure 10
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  		Fig. 10. Representative series of 1H-NMR spectra taken on carp during a 2 h recovery from exercise followed by a 4 h exposure to hypoxia. The resting/normoxic trace was taken 24 h after the initial time 0 h reading following exercise. Peaks representing creatine (Cr)/phosphocreatine, pyruvate and lactate are identified. The size of the creatine/phosphocreatine peak is not quantitative.

 

Figure 11
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  		Fig. 11. White muscle PCr (A) pHi (B), ATP (C) and lactate (D) levels from carp exposed to normoxia (N) and for up to ~8 h exposure to hypoxia. Phosphocreatine levels are normalized to the sum of PCr and Pi. PCr, ATP and lactate levels are expressed relative to normoxic controls. The black horizontal bar above the x-axis indicates the period of hypoxia exposure (PO2=20 Torr). Each point represents mean ± 1 s.d. Significant differences (P<0.05) between the resting/normoxic fish and hypoxia-exposed fish are indicated by an asterisk. From left to right, N=4, 10, 6, 3 and 4.

 

Figure 12
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  		Fig. 12. White muscle PCr (A), pHi (B), ATP (C) and lactate (D) levels from carp at exhaustion (time zero) and for up to ~8 h recovery. Phosphocreatine levels are normalized to the sum of PCr and Pi. PCr, ATP and lactate levels are expressed relative to normoxic controls. Each bar represents mean ± 1 s.d. Significant differences (P<0.05) between resting/normoxic fish and post-exercise recovery fish are indicated by an asterisk. N=3 for each point.

 

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© The Company of Biologists Ltd 2008