Subtropical hibernation in juvenile tegu lizards (Salvator merianae): insights from intestine redox dynamics.

Juvenile tegu lizards (Salvator merianae) experience gradual and mild temperature changes from autumn to winter in their habitat. This tropical/subtropical reptile enter a state of dormancy, with an 80% reduction in metabolic rate, that remains almost constant during winter. The redox metabolism in non-mammalian vertebrates that hibernate under such distinguished conditions is poorly understood. We analyzed the redox metabolism in the intestine of juvenile tegus during different stages of their first annual cycle. The effect of food deprivation (in spring) was also studied to compare with fasting during hibernation. Both winter dormancy and food deprivation caused decreases in reduced glutathione levels and glutathione transferase activity. While glutathione peroxidase and glutathione transferase activities decreased during winter dormancy, as well as glutathione (GSH) levels, other antioxidant enzymes (catalase, superoxide dismutase and glutathione reductase) remained unchanged. Notably, levels of disulfide glutathione (GSSG) were 2.1-fold higher in late autumn, when animals were in the process of depressing metabolism towards hibernation. This increased "oxidative tonus" could be due to a disruption in NADPH-dependent antioxidant systems. In dormancy, GSSG and lipid hydroperoxides were diminished by 60-70%. The results suggest that the entrance into hibernation is the main challenge for the redox homeostasis in the intestine of juvenile tegus.

Morphological and metabolic adjustments in the small intestine to energy demands of growth, storage, and fasting in the first annual cycle of a hibernating lizard (Tupinambis merianae).

Seasonal plasticity in the small intestine of neonatal tegu lizards was investigated using morphometry and analysis of enzymes involved in supplying energy to the intestinal tissue. In the autumn, the intestinal mass (Mi) was 1.0% of body mass and the scaling exponent b=0.92 indicated that Mi was larger in smaller neonates. During arousal from dormancy Mi was 23% smaller; later in spring, Mi increased 60% in relation to the autumn and the exponent b=0.14 indicated that the recovery was disproportionate in smaller tegus. During the autumn, the intestinal villi were greatly elongated; by midwinter, the Hv, SvEp, and VvEp were smaller than during the autumn (59%, 54%, 29%) and were restored to autumn levels during spring. In the active tegus, the maximum activity (Vmax) of enzymes indicated that the enterocytes can obtain energy from different sources, and possess gluconeogenic capacity. During winter, the Vmax of CS, HOAD, GDH, PEPCK was 40-50% lower in relation to the autumn and spring, while the Vmax of HK, PK, LDH, AST was unchanged. The hypoglycemia and the mucosal atrophy/ischemia during winter would prevent the enterocytes from using glucose, whereas they could slowly oxidize fatty acids released from body stores and amino acids from the tissue proteolysis to satisfy their needs of energy. Contrastingly, starvation during spring caused severe mass loss (50%); the tissue protein and the VvEp and VvLP did not change while the thickness of the muscular layer increased 51%, which suggested different effects along the length of the organ. In addition, the Vmax of the glycolytic enzymes was lower, indicating that a regulatory mechanism would spare blood glucose for vital organs during unanticipated food restriction.

Cardiac hypertrophy and structural and metabolic remodeling related to seasonal dormancy in the first annual cycle in tegu lizards.

Morpho-functional adjustments in the heart of juvenile tegu lizards (Tupinambis merianae) were analyzed at distinct seasonal periods to investigate how the demands of growth and of energy saving are reconciled during the first annual cycle. The relative ventricular mass (Mv) was 31% and 69% larger in late autumn and winter dormancy, respectively, compared to early autumn. This effect did not persist during unfed arousal, suggesting that protein accumulates in the heart during hypometabolism and is degraded on arousal. Both the hypertrophy and the atrophy were disproportionate in the largest individuals. In contrast, Mv was smaller in lizards that were starved during spring activity compared to fed lizards, this effect being larger in smaller individuals. In late autumn and winter dormancy the spongy myocardium had 8% of the section area covered by lacunary spaces, which expanded after food intake during arousal and reached 29% in spring activity together with higher density of cardiomyocytes. Total and soluble proteins per mass unity were unchanged, and maximum activities of selected enzymes suggest sustained glycolytic and aerobic capacities during hypometabolism. Results indicate that important structural adjustments occur in the heart in anticipation of dormancy, and that the protein balance in the tissue is maintained at winter temperatures ~17°C.

Metabolic scaling associated with unusual size changes during larval development of the frog, Pseudis paradoxus.

The early larvae of P. paradoxus grow large but metamorphose into relatively small frogs, the diminished post-metamorphic growth producing a marked contrast between maximum larval size and adult. Thus, O(2) uptake does not appear to limit the energy expenditure on growth processes, and unlike in other anuran larvae, may not be a surface area-related function in P. paradoxus larvae. The resting rates of metabolism (M(O(2))) and partitioning between aquatic (Mw(O(2))) and aerial O(2) uptake (Ma(O(2))) were measured on tadpoles and froglets by closed system respirometry, using water of P(O(2)) ranging from 145 to 40 mmHg. Correlative changes in body glycogen and lactate were examined by standard enzyme assays. Scaling patterns in the growth and degrowth stages were analysed on whole-body, log-transformed data using linear regressions. In normoxia, M(O(2)) was 2.1-2.5 mumol g(-1) h(-1) in the early larvae, increasing more than twofold on forelimb emergence and decreasing sharply in the froglets; M(O(2)) varies in strict proportion to body mass (M(b)), both in the growth (b=1.02) and degrowth (b=0.97) phases, according to the equation M(O(2))=aM(b)(b), where b is the scaling coefficient. Mw(O(2)) constitutes >90% of total uptake in the growth stages, increasing with b=1.02 while Ma(O(2)) increases with b=1.13; during degrowth there is a change in the pattern related to intensification of metamorphosis. Hypoxic water did not affect M(O(2)); however, in all larval stages Mw(O(2)) and Ma(O(2)) changed with a decrease in P(O(2)). At 60 mmHg, rates are more severely affected in the largest tadpoles, causing the b values for Mw(O(2)) and Ma(O(2)) to change to 0.11 and 1.44, respectively, in the growth phase. Glycogen and lactate levels increase out of proportion with body mass increase (b=2.05 and 1.47, respectively) in the growth stages, and increase anaerobic capacity in late metamorphosis. In hypoxic water, glycogen levels decrease in the growth stages and the largest tadpoles accumulate surplus lactate, possibly related to surfacing activity. Our results may reveal the consequences of size on energy demand at the tissue level in P. paradoxus larvae, indicating that air breathing must subsidise energy expenditure during larval development.

Seasonal metabolic depression, substrate utilisation and changes in scaling patterns during the first year cycle of tegu lizards (Tupinambis merianae).

The tegus increase in body mass after hatching until early autumn, when the energy intake becomes gradually reduced. Resting rates of oxygen consumption in winter drop to 20% of the values in the active season ((O(2))=0.0636 ml g(-1) h(-1)) and are nearly temperature insensitive over the range of 17-25 degrees C (Q(10)=1.55). During dormancy, plasma glucose levels are 60% lower than those in active animals, while total protein, total lipids and beta-hydroxybutyrate are elevated by 24%, 43% and 113%, respectively. In addition, a significant depletion of liver carbohydrate (50%) and of fat deposited in the visceral fat bodies (24%) and in the tail (25%) and a slight loss of skeletal muscle protein (14%) were measured halfway through the inactive period. Otherwise, glycogen content is increased 4-fold in the brain and 2.3-fold in the heart of dormant lizards, declining by the onset of arousal. During early arousal, the young tegus are still anorexic, although (O(2)) is significantly greater than winter rates. The fat deposits analysed are further reduced (62% and 45%, respectively) and there is a large decrease in tail muscle protein (50%) together with a significant increase in glycogen (2-3-fold) and an increase in plasma glucose (40%), which suggests a role for gluconeogenesis as a supplementary energy source in arousing animals. No change is detectable in citrate synthase activity, but beta-hydroxyacyl CoA dehydrogenase activities are strongly affected by season, reaching a 3-fold and 5-fold increase in the liver tissue of winter and arousing animals, respectively, and becoming reduced by half in skeletal muscle and heart of winter animals compared with late fall or spring active individuals. From hatching to late autumn, the increase of the fat body mass relatively to body mass is disproportionate (b=1.44), and the mass exponent changes significantly to close to 1.0 during the fasting period. The concomitant shift in the (O(2)) mass exponent in early autumn (b=0.75) to values significantly greater than 1.0 in late autumn and during winter dormancy indicates an allometric effect on the degree of metabolic depression related to the size of the fat stores and suggests greater energy conservation in the smaller young.