Elisabeth Debras

Effect of glucocorticoid excess on skeletal muscle and heart protein synthesis in adult and old rats

This study was carried out to analyse glucocorticoid-induced muscle wasting and subsequent recovery in adult (6-8 months) and old (18-24 months) rats because the increased incidence of various disease states results in hypersecretion of glucocorticoids in ageing. Adult and old rats received dexamethasone in their drinking water for 5 or 6 d and were then allowed to recover for 3 or 7 d. As dexamethasone decreased food intake, all groups were pair-fed to dexamethasone-treated old rats (i.e. the group that had the lowest food intake). At the end of the treatment, adult and old rats showed significant increases in blood glucose and plasma insulin concentrations. This increase disappeared during the recovery period. Protein synthesis of different muscles was assessed in vivo by a flooding dose of [13C]valine injected subcutaneously 50 min before slaughter. Dexamethasone induced a significant decrease in protein synthesis in fast-twitch glycolytic and oxidative glycolytic muscles (gastrocnemius, tibialis anterior, extensor digitorum longus). The treatment affected mostly ribosomal efficiency. Adult dexamethasone-treated rats showed an increase in protein synthesis compared with their pair-fed controls during the recovery period whereas old rats did not. Dexamethasone also significantly decreased protein synthesis in the predominantly oxidative soleus muscle but only in old rats, and increased protein synthesis in the heart of adult but not of old rats. Thus, in skeletal muscle, the catabolic effect of dexamethasone is maintained or amplified during ageing whereas the anabolic effect in heart is depressed. These results are consistent with muscle atrophy occurring with ageing.

Differential effects of insulin and dietary amino acids on muscle protein synthesis in adult and old rats

The potential roles of insulin and dietary amino acids in the regulation of skeletal muscle protein synthesis were examined in adult and old rats. Animals were fed over 1 h with either a 25% or a 0% amino acid/protein meal. In each nutritional condition, postprandial insulin secretion was either maintained or blocked with diazoxide injections. Protein synthesis in gastrocnemius and soleus muscles was assessed in vivo using the flooding dose method. Insulin suppression decreased protein synthesis in both muscles irrespective of the nutritional condition and age of the rats. Moreover, reduced insulinaemia was associated with 4E‐BP1 dephosphorylation, enhanced assembly of the 4E‐BP1−eIF4E inactive complex and hypophosphorylation of eIF4E, p70S6k and protein kinase B, key intermediates in the regulation of translation initiation and protein synthesis. Old rats did not differ from adult rats. The lack of amino acids in the meal of insulin‐suppressed rats did not result in any additional decrease in protein synthesis. In the presence of insulin secretion, dietary amino acid suppression significantly decreased gastrocnemius protein synthesis in adult but not in old rats. Amino acid suppression was associated with reduced phosphorylation of 4E‐BP1 and p70S6k in adults. Along with protein synthesis, only the inhibition of p70S6k phosphorylation was abolished in old rats. We concluded that insulin is required for the regulation of muscle protein synthesis irrespective of age and that the effect of dietary amino acids is blunted in old rats.

Effect of insulin in conjunction with glucose, amino acids and potassium on net metabolism of glucose and amino acids in the goat mammary gland

The hyperinsulinaemic euglycaemic insulin clamp technique was used to study the effect of insulin on the arterio-venous concentration differences of glucose and amino acids across the mammary gland in dairy goats. Insulin was given in conjunction with K to prevent insulin hypokalaemia. Appropriate amino acid infusion was used to blunt insulin-induced hypoaminoacidaemia or to create hyperaminoacidaemia and maintain this state under insulin treatment. Hyperaminoacidaemia alone only stimulated mammary leucine uptake but did not significantly modify the net metabolism of other amino acids and glucose. Insulin infusion at physiological level in conjunction with glucose, KCl-NaCl and amino acids failed to alter mammary uptake of glucose and essential amino acids; occasional increase in arginine extraction and decrease in tyrosine extraction were exceptions. Thus these new experimental conditions did not reveal any galactopoietic effect of insulin.

Evidence for an alteration of plasma and liver proteins response to dexamethasone in aging rats

The aim of this study was carried out to analyse the liver and plasma proteins response to dexamethasone in adult (6-8 months) and old (24 months) rats in order to ascertain the involvement of glucocorticoids in the aging process. The animals received dexamethasone (Dex) for 5 or 6 days. As Dex decreased food intake, all groups were pair fed to dexamethasone-treated old rats. The synthesis of mixed plasma and liver proteins (assessed by a flooding dose of [13C] valine) was similarly greatly improved in adult and old rats after Dex treatment. However, the level of mixed plasma proteins was only slightly increased. When specific plasma proteins were assessed, a similar increase in the concentration of albumin and alpha1 acid glycoprotein was observed in adult and old rats. By contrast, fibrinogen decreased to a greater extend in old rats and alpha2 macroglobulin became undetectable in old animals. It was concluded that the response of plasma and liver proteins to Dex was altered in old rats and may contribute to the pathogenesis of several diseases which occur during aging.

ACUTE HYPERINSULINEMIA FAILS TO CHANGE GLUT-4 CONTENT IN CRUDE MEMBRANES FROM GOAT SKELETAL MUSCLES AND ADIPOSE TISSUE

The effect of insulin on GLUT-4 protein level in samples of adipose tissue and skeletal muscles from goats was studied in vivo using an euglycemic hyperinsulinemic clamp. The clamp was maintained in conscious goats for 6 h in the presence of amino acids to prevent insulin-induced hypoaminoacidemia. GLUT-4 protein was assessed in crude membrane preparations from adipose tissue and four skeletal muscles (longissimus dorsi, tensor fasciae latae, anconeus and diaphragm) by Western blot analysis. No changes of GLUT-4 protein content were detected after 6 h of hyperinsulinemia in either adipose tissue or skeletal muscles from goats. These results suggest that insulin is not the prime factor involved in the short-term regulation of GLUT-4 protein transporter content in insulin-sensitive tissues from goats.

Amino acid supplementation prevents the loss of appetite for casein in old Lou/Cjall rats☆

A deleterious reduction of casein intake occurring earlier in males than in females had been previously observed in old Lou/Cjall rats. On the contrary, protein intake was observed to be maintained in old males when they were offered whey protein. Present studies were designed to investigate the effect of dietary casein modification on protein decrease. In two lifelong studies, male and female Lou/Cjall rats were tested every four months in order to study protein intake depending on the protein available: casein, whey protein or casein supplemented with an amino acid mixture (SC). In subsequent cross-sectional studies, young, adult, middle-aged and old rats were successively fed with casein, casein supplemented either with leucine or with alanine or with glycine. Supplementing casein with an amino acid mixture both globally increased protein intake and allowed old males to maintain a high rate of protein intake. In cross-sectional experiments, no effect of supplementation was seen in the young group. In older animals, the greatest effect was seen when casein was supplemented with alanine or glycine, independently of sex and age. We therefore, concluded that supplemented casein is more beneficial for old rats than casein alone, probably by increasing amino acid availability. We hypothesize that alanine could act through its effect on gluconeogenesis.