This study extends our earlier work (T. Raclot and R. Groscolas. J. Lipid Res. 34: 1515-1526, 1993), which showed that, under norepinephrine-stimulated lipolysis, fatty acids of rat retroperitoneal fat cells are selectively mobilized. The present study examines whether this selective mobilization of fatty acids 1) is based on their proportions in adipose tissue, 2) is a metabolic feature common to all adipose tissues, and/or 3) depends on the lipolysis-stimulating agent. Rat fat cells with two markedly different fatty acid compositions were isolated from four white adipose tissues and treated with three lipolytic agents. Fatty acid composition of in vitro released free fatty acids was compared with that of fat cell triacylglycerols, the ratio of percent in free fatty acid to percent in triacylglycerol being defined as the relative mobilization rate (RMR). The RMR of individual fatty acids was related to their molecular structure. It increased exponentially with unsaturation for a given chain length and decreased with increasing chain length for a given unsaturation. The selectivity of fatty acid mobilization was similar regardless of the fatty acid composition of adipose tissue, the tissue location, and the lipolytic agent used. Under conditions of stimulated lipolysis, the selectivity of fatty acid mobilization is therefore a general metabolic feature of adipose tissue. Fatty acids with 16-20 carbon atoms and 4 or 5 double bonds had the highest RMR (from 1.4 to > 5), whereas fatty acids with 20-22 carbon atoms and 0 or 1 double bond had the lowest RMR (from 0.3 to 0.7). For the other fatty acids, RMR was close to unity.(ABSTRACT TRUNCATED AT 250 WORDS)

The stress that might result in animals from the routine handling that most experimental studies involve, e.g., weighing, injecting, and blood sampling, is usually assumed to be minimal when the animals look quiet. However, the intensity of this stress remains largely ignored. We have developed a system that allows blood samples to be taken from freely behaving geese without entering the animal room. In these entirely undisturbed geese, the humoral indexes of stress, i.e., blood levels of catecholamines, corticosterone, and lactate, were as low or even lower than the lowest values previously reported for birds. Remarkably, the mean basal values for epinephrine and norepinephrine were 90-fold and 5-fold, respectively, below the lowest values in the literature. Stress-induced variations in pH that would have concealed detection of nutrition-induced changes in pH were eliminated. In contrast, even though the birds looked quiet during a short 5-min routine handling procedure, to which they had been accustomed for weeks, there was a dramatic increase in the level of humoral indexes of stress. These increased severalfold within only 2 min, and the return to initial values could take up to 1 h. Acid-base balance was also disrupted. Thus, in studies on animals, the absence of stress cannot be deduced from only behavioral observations. Only a system for taking blood without human interference may enable stress-free investigations.

This study tests the hypothesis that the metabolic and endocrine shift characterizing the phase II-phase III transition during prolonged fasting is related to a decrease in fatty acid (FA) oxidation. Changes in plasma concentrations of various metabolites and hormones and in lipolytic fluxes, as determined by continuous infusion of [2-3 H]glycerol and [1-14 C]palmitate, were examined in vivo in spontaneously fasting king penguins in the phase II status (large fat stores, protein sparing) before, during, and after treatment with mercaptoacetate (MA), an inhibitor of FA oxidation. MA induced a 7-fold decrease in plasma ␤-hydroxybutyrate and a 2-to 2.5-fold increase in plasma nonesterified fatty acids (NEFA), glycerol, and triacylglycerols. MA also stimulated lipolytic fluxes, increasing the rate of appearance of NEFA and glycerol by 60-90%. This stimulation might be partly mediated by a doubling of circulating glucagon, with plasma insulin remaining unchanged. Plasma glucose level was unaffected by MA treatment. Plasma uric acid increased 4-fold, indicating a marked acceleration of body protein breakdown, possibly mediated by a 2.5-fold increase in circulating corticosterone. Strong similarities between these changes and those observed at the phase II-phase III transition in fasting penguins support the view that entrance into phase III, and especially the end of protein sparing, is related to decreased FA oxidation, rather than reduced NEFA availability. MA could be therefore a useful tool for understanding mechanisms underlying the phase II-phase III transition in spontaneously fasting birds and the associated stimulation of feeding behavior.protein sparing; lipolytic fluxes; isotopic tracers; mercaptoacetate; seabirds PROLONGED FASTING IS CHARACTERIZED by the preferential utilization of lipid, with relative sparing of body protein (4, 6, 25). Previous studies indicated that protein sparing depends on the availability of lipid fuels (17). The conservation of body protein that characterizes the so-called phase II of fasting (9, 17) is no longer maintained when a lower threshold in fat stores is reached (6,17,22,25). Then a metabolic shift occurs, with a simultaneous acceleration in the catabolism of body protein and a decrease in the contribution of lipid to energy production, the signature of the so-called phase III of fasting (6,22,25). Entrance into phase III is also accompanied by hormonal changes, such as an increase in the level of circulating glucocorticoids thought to contribute to the stimulation of protein breakdown (10). How fat store availability determines body protein sparing during phase II or accelerated catabolism during phase III is not well understood. Is protein sparing during phase II linked to the availability of nonesterified fatty acids (NEFA) mobilized from adipose tissue, or does it depend on their oxidation? Arguments suggest that NEFA may specifically modulate the breakdown of myofibrillar proteins independently of their oxidation as a fuel for muscle (26). This suggestion agrees with ...

During starvation, after a short dynamic period of adaptation (phase I), a metabolic steady state is reached in which proteins are spared and lipids provide most of the energy expended [phase II (P2)]. However, protein breakdown increases dramatically once a lower threshold of body lipids is reached [phase III (P3)]. Body composition, energy intake, energy expenditure, and energy efficiency were determined in 8 groups of rats (fed, food-deprived up to P2 or P3 of starvation and refed for 3 d, 7 d, or until body mass restoration) to determine whether the kinetics of lipid and/or protein reserve recovery may be slowed down when refeeding occurs after the lipid threshold has been reached. Despite larger losses, P3 refed rats restored their body reserves as efficiently as those refed in P2. Whatever the nutritional status at the onset of refeeding, rehydration occurred first and hyperphagia played a more important role than hypometabolism in the restoration of the lost reserves. However, the pattern of body component gains was different during early refeeding. In P3 refed rats, body lipids were restored preferentially by significant contribution from endogenous lipid production. Thus, the extent of lipid depletion has important consequences for the restoration pattern of the body reserves. It depends not only on the intensity of the energy restriction (partial or total) as already demonstrated but also on the metabolic status at the onset of refeeding. These results may have significant implications on the way refeeding should be conducted after severe energy depletion.