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First published online September 19, 2008
Journal of Experimental Biology 211, 3111-3122 (2008)
Published by The Company of Biologists 2008
doi: 10.1242/jeb.019117
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AMP-activated protein kinase activity during metabolic rate depression in the hypoxic goldfish, Carassius auratus

Lindsay A. Jibb 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. Distribution of AMPK{alpha}1a (A), AMPK{alpha}1b (B) and AMPKβ1 (C) mRNA in eye, heart, brain, kidney, liver, muscle, intestine and gill in goldfish. Expression of each gene is relative to the same standard sample and absolute expression is adjusted such that the tissue with the lowest expression for each gene has an expression quantity of 1.

 

Figure 2
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  		Fig. 2. Liver [ATP] (A), [creatine phosphate] (CrP) (B) and calculated [AMPfree]/[ATP] (C) in goldfish exposed to normoxia (N; ~9.5 mg O2 l–1; open squares) and 12 h of hypoxia (~0.3 mg O2 l–1; closed squares). Horizontal broken lines through normoxia are shown as a reference. Data are means ± s.e.m. (N=5 to 8). *Significant difference from normoxia, P<0.05.

 

Figure 3
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  		Fig. 3. Liver AMPK activity (A), protein AMPK (B) and representative AMPK{alpha} western blot (C) in goldfish exposed to normoxia (N; ~9.5 mg O2l–1; open squares) and 12 h hypoxia (~0.3 mg O2l–1; closed squares). Horizontal broken lines through normoxia are shown as a reference. AMPK activity during hypoxia exposure is expressed relative to the activity observed in normoxia, which was set to 1. AMPK protein is normalized to total homogenate protein and expressed relative to the values observed in normoxia. C Bands shown on representative western blot are at ~62 kDa. Data are means ± s.e.m. (N=5 to 11). *Significant difference from normoxia, P<0.05.

 

Figure 4
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  		Fig. 4. Liver phospho-eEF2 (A), representative phospho-Thr-56-eEF2 and eEF2 western blots (B) and protein synthesis rate (C) in goldfish exposed to normoxia (N; ~9.5 mg O2l–1; open squares) and 12 h of hypoxia (~0.3 mg O2l–1; closed squares). Horizontal broken lines through normoxia are shown as a reference. Bands shown on representative western blot are at ~95 kDa. Data are means ± s.e.m. (N=5 to 8). *Significant difference from normoxia, P<0.05.

 

Figure 5
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  		Fig. 5. Muscle [ATP] (A), [creatine phosphate] (CrP) (B) and calculated [AMPfree]/[ATP] (C) in goldfish exposed to normoxia (N; ~9.5 mg O2l–1; open squares) and 12 h of hypoxia (~0.3 mg O2l–1; closed squares). Horizontal broken lines through normoxia are shown as a reference. Data are means ± s.e.m. (N=5 to 15). *Significant difference from normoxia, P<0.05.

 

Figure 6
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  		Fig. 6. Muscle AMPK activity (A), AMPK protein (B) and representative AMPK western blot (C) in goldfish exposed to normoxia (N; ~9.5 mg O2l–1; open squares) and 12 h of hypoxia (~0.3 mg O2l–1; closed squares). Data are means ± s.e.m. (N=6 to 12). See Fig. 3 caption for more detail. *Significant difference from normoxia, P<0.05.

 

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© The Company of Biologists Ltd 2008