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September 1, 1971Biochemical Journal450 citationsOpen Access

Evaluation of the isolated perfused rat hindquarter for the study of muscle metabolism

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NRNeil B. RudermanCHC.R.S. HoughtonRHR. Hems

Key Points

  • To evaluate the metabolic viability, physiological integrity, and fuel utilization patterns of an isolated perfused rat hindquarter preparation for muscle metabolism research.
  • Perfused isolated rat hindquarter preparations using a semi-synthetic medium containing aged human erythrocytes, comparing metabolic parameters in the presence and absence of skin against in vivo controls.
  • Assessed tissue high-energy phosphates (ATP, ADP, creatine phosphate), ultrastructure via electron microscopy, electrolyte shifts, and metabolic responses to insulin, bilateral sciatic nerve stimulation, and ketone bodies.
  • Muscle tissue accounted for more than 95% of total oxidative metabolism, maintaining tissue concentrations of ATP, ADP, and creatine phosphate equivalent to intact in vivo muscle after 40 minutes of perfusion.
  • Insulin stimulated a sixfold increase in glucose uptake, increased oxygen consumption by nearly 40%, and decreased glycerol release to less than half of control values.
  • Ketone bodies served as the predominant oxidative fuel in 48-hour starved states, accounting for 77% of total oxygen consumption, whereas glucose oxidation to carbon dioxide represented less than 4% of consumed oxygen.

Abstract

The metabolic integrity of a new isolated rat hindquarter preparation was studied. The hindquarter was perfused with a semi-synthetic medium containing aged human erythrocytes. More than 95% of the oxidative metabolism of the preparation was due to muscle, the remainder being due to bone, adipose tissue and, where present, skin. 2. Consumption of O(2), glucose utilization, glycerol release and lactate production were similar in the presence and in the absence of the skin, indicating that the latter contributed little to the overall metabolism of the preparation. 3. After 40min of perfusion, tissue concentrations of creatine phosphate, ATP and ADP were similar to those found in muscle taken directly from intact animals. The muscle also appeared normal under the electron microscope. 4. The hindquarter did not lose K(+) to the medium during a 30min perfusion. In the presence of insulin it had a net K(+) uptake. 5. Insulin caused a sixfold increase in glucose uptake, stimulated O(2) consumption by nearly 40% and depressed glycerol release to less than half the control value. 6. Bilateral sciatic-nerve stimulation caused severalfold increases in O(2) consumption and lactate production. In the absence of insulin nerve stimulation also enhanced glucose uptake; in the presence of insulin it did not further increase the already high rate of glucose uptake. 7. Rates of lactate production and O(2) consumption of the rat hindquarter in vivo and the isolated perfused hindquarter were very similar. 8. Ketone bodies were a major oxidative fuel in vivo of the hindquarter of a rat starved for 2 days. If the acetoacetate and 3-hydroxybutyrate removed by the tissue were completely oxidized, they would have accounted for 77% of the O(2) consumption. 9. Acetoacetate accounted for 84% of the ketone bodies removed by the hindquarter in vivo even though its arterial concentration was half that of 3-hydroxybutyrate. 10. Similar rates of acetoacetate and 3-hydroxybutyrate utilization were observed in the perfused hindquarter. 11. Acetoacetate utilization by the perfused hindquarter was not diminished by the addition of either oleate or insulin to the perfusate. 12. Oxidation of glucose to CO(2) accounted for less than 4% of the O(2) consumed by the perfused hindquarter in both the presence and the absence of insulin. 13. The results indicate that the isolated perfused hindquarter is a useful tool for studying muscle metabolism. They also suggest that ketone bodies, if present in sufficient concentration, are the preferred oxidative fuel of resting muscle.

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Cite This Study

Ruderman et al. (1971) studied this question.

synapsesocial.com/papers/6a1777e44f2b3115b01296cchttps://doi.org/10.1042/bj1240639
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