Hormonal effects on fatty-acid synthase in cultured fetal rat lung; induction by dexamethasone and inhibition of activity by triiodothyronine.

We previously reported that administration of dexamethasone to the pregnant dam increased the activity of fatty-acid synthase (EC 2.3.1.85) in fetal rat lung and that this effect was reduced when triiodothyronine (T3) was also administered. To determine whether the hormones act directly on the lung, we examined their effects in organ culture. Explants of 18-day and 19-day fetal rat lung were cultured with 100 nM dexamethasone or 100 nM T3, the two hormones together or no hormone at all for 48 h, after which fatty-acid synthase was assayed. Dexamethasone increased fatty-acid synthase activity at both gestational ages. T3 alone had no effect on 18-day, but decreased the activity in 19-day explants by 20%. T3 reduced the stimulatory effect of dexamethasone from 177% to 102% and from 61% to 22% in 18- and 19-day explants, respectively. The effects of dexamethasone and T3 were concentration dependent, with EC50 (concentration achieving 50% of the maximum effect) values of 0.65 nM and approx. 25 nM, respectively. This dexamethasone EC50 is lower than the reported Kd for dexamethasone binding, but the T3 EC50 is considerably higher than its reported Kd. The physiological significance of the T3 effect is, therefore, not clear. The effect of dexamethasone was not apparent until at least 12 h after exposure to the hormone and it was abolished by actinomycin D. Immunoprecipitation with antibody against rat liver fatty-acid synthase showed that there was more fatty-acid synthase in the dexamethasone-treated than in the control cultures. The potency order of glucocorticoids in stimulating fatty-acid synthase was similar to that previously reported for specific nuclear glucocorticoid binding. These data show that dexamethasone and T3 act directly on the fetal lung and that the stimulatory effect of the glucocorticoid on fatty-acid synthase is due to new protein synthesis.

Effects of glucocorticoid and thyroid hormones on regulatory enzymes of fatty acid synthesis and glycogen metabolism in developing fetal rat lung.

Although glucocorticoid and thyroid hormones are known to act synergistically to stimulate surfactant production, they have opposite effects on other parameters of fetal lung maturation. We recently reported that the developmental increases in de novo fatty acid synthesis and glycogen accumulation in fetal rat lung were accelerated by dexamethasone but prevented by triiodothyronine and that the dexamethasone-induced increases were diminished when the two hormones were administered together. We have now examined the effects of maternal administration of these hormones on activities of enzymes of lung fatty acid synthesis and glycogen metabolism in the rat. There was a developmental increase in fatty-acid synthase activity between 19 and 21 days gestation. This activity was increased by dexamethasone but decreased by triiodothyronine. When the two hormones were administered together the stimulatory effect of dexamethasone was decreased from 56% to 29%. The stimulatory effect on fatty-acid synthase was also observed in fetal lung explants cultured in the presence of dexamethasone. This shows that the effect of the hormone was directly on the fetal lung. Dexamethasone had no effect on liver fatty-acid synthase. There was a developmental decrease in acetyl-CoA carboxylase activity but it was not affected by the hormones. These data show that the developmental and hormone-induced changes in fetal lung de novo fatty acid synthesis are mediated by fatty-acid synthase. Although there were developmental changes in fetal lung 6-phosphofructokinase, glycogen synthase and glycogen phosphorylase activities, these enzymes were not affected by the hormones.

Dietary dependent distribution of acetyl CoA carboxylase between cytoplasm and mitochondria of rat liver.

Biotinyl proteins in cytoplasm and mitochondria of rat liver were examined by fluorography and the quantity of acetyl CoA carboxylase was determined after sodium dodecyl sulfate-denatured proteins were incubated with [14C] methyl avidin and separated by polyacrylamide gel electrophoresis. Results show that one-half of the total acetyl CoA carboxylase in liver of fed rats was associated with mitochondria in a relatively inactive form. Fasting shifted the distribution of the enzyme toward the mitochondrial fraction and refeeding previously fasted rats shifted the distribution towards cytoplasm. Thus, acetyl CoA carboxylase can be added to the list of ambiquitous enzymes whose subcellular distribution varies with physiological conditions.

Molecular weights of subunits of acetyl CoA carboxylase in rat liver cytoplasm.

Monomeric [14C] methyl avidin was shown to bind to sodium dodecyl sulfate-denatured biotinyl proteins and remain bound through polyacrylamide gel electrophoresis which allowed their detection by fluorography. This method was used to show that purified rat liver acetyl CoA carboxylase contained two high molecular weight forms of the enzyme (MR = 241,000 and 252,000) while rapidly prepared, crude rat liver cytoplasm contained two larger molecular weight (MR = 257,000 and 270,000) forms. Thus, the enzyme had undergone substantial proteolysis during purification. The crude enzyme preparation also contained a smaller biotinyl protein (MR = 141,000) which is likely a proteolytic product of the larger forms of acetyl CoA carboxylase.