Synergy between angiotensin and aldosterone in evoking sodium appetite in baboons.

The synergy between ANG II and aldosterone (Aldo) in the induction of salt appetite, extensively studied in rats, has been tested in baboons. ANG II was infused intracerebroventricularly at 0.5 or 1.0 microg/h; Aldo was infused subcutaneously at 20 microg/h. Separate infusions over 7 days had no significant effect on the daily intake of 300 mM NaCl. Concurrent infusions, however, increased daily NaCl intake approximately 10-fold and daily water intake approximately 2.5-fold. In addition, the combined infusions caused 1) a reduction in daily food intake, 2) changes in blood composition indicative of increased vasopressin release, and 3) changes of urinary excretion rates of cortisol and Aldo indicative of increased ACTH release. Arterial blood pressure, measured in two baboons, rose during concurrent ANG II and Aldo treatment. These results indicate a potent synergy between central ANG II and peripheral Aldo in stimulating salt appetite in baboons. At the same time, other ANG II-specific brain mechanisms concerned with water intake, food intake, vasopressin release, ACTH release, and blood pressure regulation appear to have been activated by the same type of synergy. These central enhancement processes have never been previously demonstrated in primates.

Ingestive responses to administration of stress hormones in baboons.

Experimental stress and the administration of the stress hormone ACTH have been reported to stimulate sodium appetite in many nonprimate species. Experiments were conducted to determine whether prolonged intracerebroventricular infusions of the neuropeptides corticotropin-releasing factor (CRF) and urocortin (Ucn), or systemic administration of ACTH, affected ingestive behaviors in a nonhuman primate, the baboon. Intracerebroventricular infusions of CRF or Ucn significantly decreased daily food intake. The decrease with Ucn continued into the postinfusion period. These infusions did not alter daily water intake. Daily voluntary intake of 300 mM NaCl solution was not increased, and there was evidence of reductions on days 2-4 of the infusions. Intramuscular injections of porcine ACTH or synthetic ACTH (Synacthen) for 5 days did not affect daily NaCl intake, although the doses were sufficient to increase cortisol secretion and arterial blood pressure. Sodium depletion by 3 days of furosemide injections did induce a characteristic sodium appetite in the same baboons. These results demonstrate the anorexigenic action of CRF and Ucn in this primate. Also, CRF, Ucn, and ACTH did not stimulate sodium appetite at the doses used.

Possible contribution of brain angiotensin III to ingestive behaviors in baboons.

Recent experiments with specific aminopeptidase inhibitors in rats have strengthened earlier proposals that ANG III may be an important regulatory peptide in the brain. Central mechanisms regulating blood pressure, ingestive behaviors, and vasopressin release could be involved. Arguments in favor of a role for ANG III depend, in part, on the efficacy of ANG III as an agonist. These first studies in primates tested whether ANG III stimulates ingestive behaviors in baboons. Intracerebroventricular (ICV) infusions of ANG III were as potent as ANG II in stimulating water drinking and intake of NaCl solution. On the basis of this criterion and consistent with findings in rats, ANG III could be a main effector peptide in the regulation of ingestive behaviors in a primate.

Intraventricular neuropeptide Y decreases need-induced sodium appetite and increases pica in rats.

Neuropeptide Y (NPY) is a potent endogenous stimulator of food intake. In addition to stimulating increased food intake, when paired with a novel-flavored solution, NPY produces an aversion to that flavor. Hence, exogenous NPY elicits 2 seemingly opposing behaviors, increased feeding and the formation of a conditioned taste aversion. One interpretation of these data is that NPY produces some form of malaise or visceral illness. NPY's orexigenic and malaise-inducing properties were tested in rats with 2 measures sensitive to malaise, increased kaolin consumption (pica behavior) and failure to express need-induced sodium intake. Administration of NPY resulted in increased food intake, increased kaolin consumption, and decreased need-induced sodium intake. These data support the hypothesis that exogenous NPY has both orexigenic and malaise-inducing properties.

Effect of exogenous cholesterol and dithiothreitol on the activity of human liver microsomal acyl-coenzyme A:cholesterol acyltransferase (ACAT).

Acyl-coenzyme A:cholesterol acyltransferase (ACAT) is the intracellular enzyme responsible for the esterification of cholesterol with long-chain fatty acyl-CoA derivatives and has been implicated in atherosclerosis and gallstone disease. The effects of exogenous cholesterol and dithiothreitol (DTT) on the ACAT activity of human liver microsomes have been determined. Pre-incubation of microsomes with exogenous cholesterol gave a marked stimulation of activity. Experiments with [3H]cholesterol and [14C]oleoyl-CoA indicated the time course of equilibration of exogenous with endogenous cholesterol as ACAT substrates, and showed that ACAT activity could be accurately measured using [3H]cholesterol/Tween 80, providing that the concentration of endogenous microsomal cholesterol was also determined. Pre-incubation of liver microsomes for 90 min in the presence of 2 mmol/l DTT and exogenous cholesterol/Tween 80 resulted in a 60% reduction in ACAT activity, compared with the corresponding activity when DTT was omitted. If microsomes were pre-incubated with DTT prior to the pre-incubation with exogenous cholesterol/Tween 80, an 85-90% reduction in ACAT activity occurred. In contrast, pre-incubation of microsomes with DTT in the absence of exogenous cholesterol/Tween 80 (only endogenous cholesterol present) resulted, initially in a stimulation of ACAT activity; on further pre-incubation, activity returned to control levels. These results indicate that the supply of cholesterol to the enzyme active site is an important factor in ACAT assays in vitro and that DTT has a major effect on this process, suggesting that these factors may be important in controlling ACAT activity in vivo.

Fatty acid ethyl ester synthesis by human liver microsomes.

Fatty acid ethyl esters are a family of non-oxidative metabolites of ethanol present in many tissues after ethanol consumption. In this report we demonstrate the existence in human liver of an acyl-CoA:ethanol acyltransferase activity which may be responsible in part for the synthesis of these compounds in vivo. The effects of oleoyl-CoA and ethanol concentrations, presence or absence of bovine serum albumin and detergent, pH and enzyme concentration on this activity have been determined. Acyl-CoA:ethanol acyltransferase activity is localised in the membrane-bound fraction. Using inhibitors directed against related enzyme activities, it has been shown that the activity is not related to serine-dependent carboxylesterases or acyl-CoA:cholesterol acyltransferase, but tht it may be associated with acyl-CoA hydrolase activity. We have also compared acyl-CoA:ethanol acyltransferase activity with fatty acid ethyl ester synthase activity in microsomes and cytosol from the same liver. Our data indicate that these activities are comparable in vitro (on a unit/g liver basis), and suggest that both may be significant in vivo.

Intraventricular insulin and the level of maintained body weight in rats.

To determine whether central insulin administration lowers the level around which body weight is regulated, insulin (6 mU/day) or saline was infused into the third ventricles of four groups of rats. One insulin-infused and one saline-infused group were food-deprived for 3 days and were then returned to an ad lib feeding schedule. The other two groups were maintained on ad lib feeding throughout. Insulin-fused food-deprived rats. In ad lib fed rats, insulin caused a significant reduction of food intake and weight relative to saline-infused controls. When formerly food-deprived rats were returned to ad lib feeding, they gained weight, and this was significantly more pronounced in the saline-infused than the insulin-fused group. The body weights of the two insulin-infused groups converged on a value approximately 9% below the average of the two saline infused groups, with one group increasing its weight and the other decreasing its weight to achieve that weight. These findings suggest that the third-ventricular infusion of insulin does not incapacitate the rats and that they can alter their food intake either upward or downward to attain a new weight. The results are also consistent with the hypothesis that direct administration of insulin into the brain determines the level of weight maintained by the animal.