INFLUENCE OF DIETARY-PROTEIN, ENERGY AND CORTICOSTEROIDS ON PROTEIN-TURNOVER, PROTEOGLYCAN SULFATION AND GROWTH OF LONG-BONE AND SKELETAL-MUSCLE IN THE RAT

1. We report here the extent to which changes in protein turnover contribute to the previously described inhibition of growth of rat tibial length and skeletal muscle mass in response to protein deficiency [1], energy restriction and corticosterone treatment [2]. Measurements of S-35 uptake in vivo also enabled the qualitative pattern of changes in proteoglycan synthesis in bone and muscle to be established. 2. Protein deficiency was examined by ad libitum feeding of 20%, 7%, 3.5% and 0.5% protein diets with measurements at 1, 3 and 7 days (all diets), and 14 and 21 days (0.5% protein). In bone this induced delayed inhibition of tibial growth with parallel inhibition of protein synthesis, as measured by the phenylalanine flooding dose method. This was mediated by reductions in both ribosomal capacity (RNA/protein ratio) and activity (protein synthesis/RNA) in the 0.5% protein group. The pattern of inhibition of proteoglycan sulphation, measured as S-35 uptake 60 min after injection of a tracer dose of labelled sulphate, was similar to that of protein synthesis. 3. In muscle there was an intermediate graded inhibition of protein synthesis by protein deficiency, mediated by reductions in both ribosomal capacity and activity in the 0.5% protein group, which preceded growth inhibition in the 7% and 3.5% groups, and which was progressive with time. Transient increases in proteolysis contributed to the growth inhibition is some groups, but the rate fell eventually in the 0.5% group. The pattern of response of proteoglycan sulphation differed from protein synthesis with a delayed inhibition, but with subsequent marked reduction. 4. Energy restriction was induced by diets fed for 4 or 8 days at 75%, 50% and 25% ad libitum intakes with protein intakes held constant, and corticosterone treatment involved a dose of 10 mg day(-1) 100(-1) g (subcutaneous) with ad libitum feeding. In bone this induced a pattern of length growth inhibition which was dissociated from inhibition of protein synthesis in the moderately restricted (75% and 50%) groups. Only in the 25% group and in the 8 day corticosterone group was protein synthesis inhibited, through reductions in ribosomal capacity and activity. S-35 uptake was also dissociated from growth inhibition, with reduced S-35 uptake observed only after corticosterone treatment or 8 days of the 50% or 25% diets. 5. In muscle the energy restriction and corticosterone treatment induced parallel inhibitions of growth and protein synthesis, mediated by similar graded reductions in the RNA/protein ratios and in the 25% group in the K-RNA. Proteolysis was unchanged in all except the 4-day corticosterone group (elevated by 25%) and the day 8 25% group (elevated by 40%) and corticosterone group (elevated by 60%). S-35 uptake was inhibited in parallel to muscle growth and protein synthesis. 6. These data show that inhibition of protein synthesis and S-35 uptake is an invariable element of muscle growth inhibition, and a usual but not invariable element of bone growth inhibition. Partial correlation analysis of the interactions between dietary protein, bone growth and muscle protein and proteoglycan synthesis shows that bone growth (as indicated by epiphyseal cartilage width) is significantly correlated with muscle protein synthesis and especially S-35 uptake, suggesting that the regulation of muscle growth by passive stretch consequent an bone lengthening includes muscle connective tissue growth as an important target.