Proteins in the degenerative retina of C3H mice: deficiency of a cyclic-nucleotide phosphodiesterase and opsin

Abstract— The protein patterns from retinae of mice with inherited blindness (C3H/HeJ) were compared with those from normal (DBA/1J) retinae during postnatal maturation. The number of protein bands, as determined by polyacrylamide gel electrophoresis in sodium dodecyl sulphate, increased in the normal retina with development and, by adulthood, 30 bands of protein were observed, with molecular weights ranging from 13,500 to 105,000 daltons. Four bands of retinal protein were suggested to be histones, and one was identified as tubulin. The patterns of protein bands from the immature C3H retinae were similar to those from the DBA retinae but, by adulthood, the pattern from the C3H retina was deficient in three bands of protein, two of which were identified as opsin and a cyclic-nucleotide phosphodiesterase.     

RNA and DNA in developing retinae: comparison of a normal with the degenerating retinae of C3H mice

Abstract— The Nucleic acids were measured in developing retinae of normal (DBA) mice and those afflicted with an inherited degenerative disease (C3H). The content of RNA in normal and C3H retinae increased to a maximum at 5 days of postnatal age. Thereafter, that of C3H retinae declined to a value lower than the normal. The content of DNA in normal and C3H retinae was maximal at 10 and 5 days of postnatal age, respectively. By 20 days, it declined in both retinae to nearly adult values. The DNA/RNA ratio of normal adult retinae was about 3, while that of C3H adult retinae was nearly 1–5. It is proposed that the photoreceptor cells possess a smaller cytoplasmic volume and a larger DNA/RNA ratio than the cells of the inner retina. The loss of DNA in developing normal and C3H retinae appears to result from cellular death. It was calculated that approximately 1 million cells in normal and 6 million cells in C3H retinae disappear during development. Cellular death in C3H retinae may not be restricted to the photoreceptor population.     

Metabolic responses to exhaustive exercise change markedly during the protracted non-trophic spawning migration of the lamprey <Emphasis Type="Italic">Geotria australis</Emphasis>

Adults of the Southern hemisphere lamprey Geotria australis were subjected to an exercise/recovery regime at the commencement and end of their 12–15 month non-trophic, upstream spawning migration. In early (immature) migrants and pre-spawning females, muscle glycogen was markedly depleted during exercise, but became rapidly replenished. As muscle lactate rose during exercise and peaked 1–1.5 h into the recovery period, and therefore after muscle glycogen had become replenished, it cannot be the direct source for that replenishment. However, both plasma lactate and glycerol (but not muscle glycerol and glucose) rose sharply during exercise and then declined markedly during the first 0.5 h of recovery and thus exhibited the opposite trend to that of muscle glycogen, implying that these limited pools of glycogenic precursors contribute to glycogen replenishment. Although plasma glucose rose following exercise, and consequently could also be a precursor for muscle glycogen replenishment, it remained elevated even after muscle glycogen had become replenished. While resting pre-spawning females and mature males retained high muscle glycogen concentrations, this energy store became permanently depleted in females during spawning. In mature males, muscle glycogen remained high and lactate low during the exercise/recovery regime, whereas muscle glycerol declined precipitously during exercise and then rose rapidly. In summary, vigorous activity by G. australis is fuelled extensively by anaerobic metabolism of glycogen early in the spawning run and by pre-spawning females, but by aerobic metabolism of its energy reserves in mature males.     

Glucose metabolism in the newborn rat. Temporal studies in vivo

1. The concentrations of plasma d-glucose, l-lactate, free fatty acids and ketone bodies and of liver glycogen were measured in caesarian-delivered newborn rats at time-intervals up to 4h after delivery. Glucose and lactate concentrations decreased markedly during the first hours after delivery, but there was a delay of 60-90min before significant glycogen mobilization occurred. 2. The specific radioactivity of plasma d-glucose was measured as a function of time for up to 75min after the intraperitoneal injection of d-[6-(14)C]glucose and d-[6-(3)H]glucose into caesarian-delivered rats at 0, 1 and 2h after delivery. Calculations revealed that there was an appreciable rate of glucose formation at all ages studied, but immediately after delivery this was exceeded by the rate of glucose utilization. Around 2h post partum the rate of glucose utilization decreased dramatically and this coincided with a reversal of the immediately postnatal hypoglycaemia. 3. The specific radioactivity of plasma l-lactate and the incorporation of (14)C into plasma d-glucose and liver glycogen was measured as a function of time after the intraperitoneal injection of l-[U-(14)C]lactate into rats immediately after delivery. The logarithm of the specific radioactivity of plasma l-[U-(14)C]lactate decreased linearly with time for at least 60min after injection and the calculated rate of lactate utilization exceeded the rate of lactate formation. 4. (14)C incorporation into plasma d-glucose was maximal from 30-60min after injection of l-[U-(14)C]lactate and the amount incorporated at 60min was 23% of that present in plasma lactate. Although (14)C was also incorporated into liver glycogen the amount was always less than 3% of that present in plasma glucose. 5. The results are discussed in relationship to the adaptation of the newly born rat to the extra-uterine environment and the possible involvement of gluconeogenesis at this time before feeding is established.     

Glycogen metabolism in the rat retina

It has been reported that glycogen levels in retina vary with retinal vascularization. However, the electrical activity of isolated retina depends on glucose supply, suggesting that it does not contain energetic reserves. We determined glycogen levels and pyruvate and lactate production under various conditions in isolated retina. Ex vivo retinas from light- and dark-adapted rats showed values of 44 +/- 0.3 and 19.5 +/- 0.4 nmol glucosyl residues/mg protein, respectively. The glycogen content of retinas from light-adapted animals was reduced by 50% when they were transferred to darkness. Glycogen levels were low in retinas incubated in glucose-free media and increased in the presence of glucose. The highest glycogen values were found in media containing 20 mm of glucose. A rapid increase in lactate production was observed in the presence of glucose. Surprisingly, glycogen levels were the lowest and lactate production was also very low in the presence of 30 mm glucose. Our results suggest that glycogen can be used as an immediate accessible energy reserve in retina. We speculate on the possibility that gluconeogenesis may play a protective role by removal of lactic acid.     

Gluconeogenic pathway in liver and muscle glycogen synthesis after exercise

To determine whether prior exercise affects the pathways of liver and muscle glycogen synthesis, rested and postexercised rats fasted for 24 h were infused with glucose (200 mumol.min-1.kg-1 iv) containing [6-3H]glucose. Hyperglycemia was exaggerated in postexercised rats, but blood lactate levels were lower than in nonexercised rats. The percent of hepatic glycogen synthesized from the indirect pathway (via gluconeogenesis) did not differ between exercised (39%) and nonexercised (36%) rats. In red muscle, glycogen was synthesized entirely by the direct pathway (uptake and phosphorylation of plasma glucose) in both groups. However, only approximately 50% of glycogen was formed via the direct pathway in white muscle of exercised and nonexercised rats. Therefore prior exercise did not alter the pathways of tissue glycogen synthesis. To further study the incorporation of gluconeogenic precursors into muscle glycogen, exercised rats were infused with either saline, lactate (100 mumol.min-1.kg-1), or glucose (200 mumol.min-1.kg-1), containing [6-3H]glucose and [14C(U)]lactate. Plasma glucose was elevated one- to twofold and three- to fourfold by lactate and glucose infusion, respectively. Plasma lactate levels were elevated by about threefold during both glucose and lactate infusion. Glycogen was partially synthesized via an indirect pathway in white muscle and liver of glucose- or lactate-infused rats but not in saline-infused animals. Thus participation of an indirect pathway in white skeletal muscle glycogen synthesis required prolonged elevation of plasma lactate levels produced by nutritive support.     

Glucose metabolism in the newborn rat. Hormonal effects in vivo

1. The concentrations of liver glycogen and plasma d-glucose were measured in caesarian-delivered newborn rats at time-intervals up to 3h after delivery after treatment of the neonatal rats with glucagon, dibutyryl cyclic AMP, cortisol or cortisol+dibutyryl cyclic AMP. Glycogenolysis was promoted by glucagon or dibutyryl cyclic AMP in the third hour after birth but not at earlier times. Cortisol and dibutyryl cyclic AMP together (but neither agent alone) promoted glycogenolysis in the second hour after birth, but no hormone combination was effective in the first postnatal hour. 2. The specific radioactivity of plasma d-glucose was measured as a function of time for up to 75 min after the intraperitoneal injection of d-[6-(14)C]glucose and d-[6-(3)H]glucose into newborn rats at delivery and after treatment with glucagon or actinomycin D. Glucagon-mediated hyperglycaemia at this time was due to an increased rate of glucose formation and a decreased rate of glucose utilization. Actinomycin D prevented glucose formation and accelerated the rate of postnatal hypoglycaemia. 3. The specific radioactivity of plasma l-lactate and the incorporation of (14)C into plasma d-glucose was measured as a function of time after the intraperitoneal injection of l-[U-(14)C]lactate into glucagon- or actinomycin D-treated rats immediately after delivery. The calculated rates of lactate formation were unchanged by either treatment, but lactate utilization was stimulated by glucagon administration. Glucagon stimulated and actinomycin D diminished (14)C incorporation into plasma d-glucose. 4. The factors involved in the initiation of glycogenolysis and gluconeogenesis in the rat immediately after birth are discussed.     

CYCLIC-NUCLEOTIDE PHOSPHODIESTERASE : An Early Defect in Inherited Retinal Degeneration of C3H Mice

Cyclic nucleotides have been implicated in the differentiation and function of the vertebrate retina. In the normal retina of DBA mice, the specific activity of cyclic-nucleotide phosphodiesterase (PDE), with cyclic-AMP as the substrate (cAMP-PDE), increases eightfold between the 6th and 20th postnatal day. Kinetic analysis of retinae from newborn mice reveals a PDE with a single Michaelis constant (K_m) value for cyclic-AMP (low K_m-PDE). After the 6th postnatal day, a second PDE with a high K_m for cyclic-AMP (high K_m-PDE) can be demonstrated. The appearance and increasing activity of the high K_m-PDE coincides with the differentiation and growth of photoreceptor outer segments. Additionally, the high K_m-PDE is shown by microchemical techniques to be concentrated in the photoreceptor cell layer and the low K_m-PDE within the inner layers of the normal retina. In C3H mice afflicted with an inherited degeneration of the photoreceptor layer, the postnatal increase in the specific activity of cAMP-PDE is substantially lower than in the normal retina. The postnatal increase in the specific activity of cAMP-PDE in two regions of the brain of C3H mice is the same as in the normal strain. A deficiency in high K_m-PDE activity in the C3H retina is evident on the 7th postnatal day, when the activity of low K_m-PDE, photoreceptor morphology, and rhodopsin content of these retina are essentially normal. In the adult C3H retina, the PDE activity with cyclic-GMP and cyclic-UMP as substrates is significantly below that of the normal retina. These data indicate that an alteration in cyclic-AMP metabolism occurs before photoreceptor cell degeneration in the retinae of C3H mice.     

Isolation and Identification of l-Deoxy-l-(L-asparagino)-D-fructose Formed in the Autoclaved Culture Medium

To elucidate the effect of nutrition during induction on peripheral muscle responsiveness to insulin, the incorporation of radiolabeled glucose to glycogen and the uptake of radiolabeled deoxyglucose were studied in isolated diaphragms from the fetuses of normal and diabetic pregnant rats in vitro. Basal- and insulin-stimulated incorporation of [1-¹⁴C]glucose into diaphragm glycogen were greater in the fetuses of diabetic mothers (IDM) than in normal fetuses, but there was no difference in the degree of stimulation by insulin of labeled glucose into glycogen between normal fetuses and IDM. Diaphragms from normal fetuses and IDM had the same basal uptake of 2-deoxy-[1-³H]glucose as well as insulin-stimulated uptake. Consequently the sensitivity of glucose uptake to insulin was similar both in normal fetuses and IDM. These data indicate that glucose utilization (incorporation of labeled glucose into glycogen) was increased in IDM, but that the response of glucose uptake and glycogenesis to insulin was not altered.     

Alterations in cyclic AMP metabolism associated with photoreceptor cell degeneration in the C3H mouse

Previous studies have shown an abnormality in cyclic nucleotide phosphodiesterase activity in the retina of mice (C3H/HeJ) with an inherited degeneration of the photoreceptor layer. Adenyl cyclase activity and cyclic AMP content have been measured in C3H retina and compared with that in normal retina (DBA/1J) during postnatal maturation, to assess the influence of the mutation upon cyclic AMP metabolism. Adenyl cyclase activity increases normally for the first 7 days of age; thereafter, it becomes greater than normal. Cyclic AMP becomes obviously abnormal after 10 days of age. The elevated levels of cyclic AMP persist in the surviving cells of the inner layers of the adult C3H retina. Adenyl cyclase activity and cyclic AMP content are concentrated in the inner layers of the normal retina, while the photoreceptor layer has only a very low level of enzyme activity and cyclic AMP. The data suggest that the synthesis of cyclic AMP in the inner layers of C3H retina is significantly enhanced, during the period of postnatal development in which the photoreceptor cells have begun to degenerate.     

Effect of fasting on carbohydrate metabolism in frugivorous bats (Artibeus lituratus and Artibeus jamaicensis)

The compensatory changes of carbohydrate metabolism induced by fasting were investigated in frugivorous bats, Artibeus lituratus and Artibeus jamaicensis. For this purpose, plasma levels of glucose and lactate, liver and muscle glycogen content, rates of liver gluconeogenesis and the activity of related enzymes were determined in male bats. After a decrease during the first 48 h of fasting, plasma glucose levels remained constant until the end of the experimental period. Plasma lactate levels, extremely high in fed bats, decreased after 48 h of fasting. Similarly, liver glycogen content, markedly high in fed animals, was reduced to low levels after 24 h without food. Muscle glycogen was also reduced in fasted bats. The expected increase in liver gluconeogenesis during fasting was observed after 48 h of fasting. The activities of liver glucose-6-phosphatase and fructose-1,6-bisphosphatase were not affected by food withdrawn. On the other hand, fasting for 24 h induced an increase in the activity of liver cytosolic phosphoenolpyruvate carboxykinase. The data indicate that liver gluconeogenesis has an important role in the glucose homeostasis in frugivorous bats during prolonged periods of food deprivation. During short periods of fasting liver glycogenolysis seems to be the main responsible for the maintenance of glycemia.     

Nutrition during brain activation: does cell-to-cell lactate shuttling contribute significantly to sweet and sour food for thought?

Functional activation of astrocytic metabolism is believed, according to one hypothesis, to be closely linked to excitatory neurotransmission and to provide lactate as fuel for oxidative metabolism in neighboring neurons. However, review of emerging evidence suggests that the energetic demands of activated astrocytes are higher and more complex than recognized and much of the lactate presumably produced by astrocytes is not locally oxidized during activation. In vivo activation studies in normal subjects reveal that the rise in consumption of blood-borne glucose usually exceeds that of oxygen, especially in retina compared to brain. When the contribution of glycogen, the brain's major energy reserve located in astrocytes, is taken into account the magnitude of the carbohydrate-oxygen utilization mismatch increases further because the magnitude of glycogenolysis greatly exceeds the incremental increase in utilization of blood-borne glucose. Failure of local oxygen consumption to equal that of glucose plus glycogen in vivo is strong evidence against stoichiometric transfer of lactate from astrocytes to neighboring neurons for oxidation. Thus, astrocytes, not nearby neurons, use the glycogen for energy during physiological activation in normal brain. These findings plus apparent compartmentation of metabolism of glycogen and blood-borne glucose during activation lead to our working hypothesis that activated astrocytes have high energy demands in their fine perisynaptic processes (filopodia) that might be met by glycogenolysis and glycolysis coupled to rapid lactate clearance. Tissue culture studies do not consistently support the lactate shuttle hypothesis because key elements of the model, glutamate-induced increases in glucose utilization and lactate release, are not observed in many astrocyte preparations, suggesting differences in their oxidative capacities that have not been included in the model. In vivo nutritional interactions between working neurons and astrocytes are not as simple as implied by "sweet (glucose-glycogen) and sour (lactate) food for thought."     

Muscle glycogen, lactate and glycerol-3-phosphate concentrations of larval and young adult lampreys in response to exercise

When stimulated, the ammocoetes (larvae) of Geotria australis swim continuously at a moderate rate for only approximately 20 min, whereas the downstream migrants (young adults) of this species did not become exhausted following similar swimming activity over the same period. Mean concentrations of muscle glycogen in ammocoetes declined during exercise, but returned to resting levels within 30 min of recovery, whereas those in young adults changed little during the corresponding periods. Moreover, muscle lactate concentrations of ammocoetes rose markedly during exercise and the first 30 min of recovery, before declining significantly, while those of young adults remained similar during and immediately after exercise. Calculations, using the glycogen and lactate concentrations immediately after exercise, suggest that during exercise glycogen is, to some extent, utilised anaerobically (approx. 24%) by ammocoetes, but only aerobically by young adults. Furthermore, since young adults used only a small amount of glycogen, they presumably metabolised triacylglycerol aerobically to produce energy. Muscle glycerol-3-phosphate levels were far higher prior to and immediately after exercise in downstream migrants than in ammocoetes and then declined precipitously. The above trends in muscle glycogen and lactate of larval G. australis parallels, to some degree, those recorded by other workers for upstream migrant Petromyzon marinus that had been exercised to exhaustion.     

Changes in plasma thyroxine,triiodothyronine and cortisol associated with starvation in rainbow trout (Salmo gairdneri)

Synopsis Chronically starved rainbow trout (Salmo gairdneri) showed a significant fall in liver size, total liver glycogen, liver glycogen concentration and plasma glucose levels. Liver lipid concentration did not differ significantly from controls although total liver lipid reserves fell during the first 40 days of starvation but had partly recovered after 65 days of starvation. Plasma cortisol and T3 levels did not show consistent changes concomitant with food deprivation over the 65 day period of the experiment. However, plasma T4 levels in fish starved for 40 or 65 days were significantly lower than comparably fed animals. The involvement of T4 in intermediate metabolic processes in salmonids is discussed.     

Lactate metabolism in the perfused rat hindlimb.

A preparation of isolated rat hindleg was perfused with a medium consisting of bicarbonate buffer containing Ficoll and fluorocarbon, containing glucose and/or lactate. The leg was electrically prestimulated to deplete partially muscle glycogen. The glucose was labelled uniformly with 14C and with 3H in positions 2, 5 or 6, and lactate uniformly with 14C and with 3H in positions 2 or 3. Glucose carbon was predominantly recovered in glycogen, and to a lesser extent in lactate. The 3H/14C ration in glycogen from [5-3H,U-14C]- and [6-3H,U-14C]-glucose was the same as in glucose. Nearly all the utilized 3H from [2-3H]glucose was recovered as water. Insulin increased glucose uptake and glycogen synthesis 3-fold. When the muscle was perfused with a medium containing 10 mM-glucose and 2 mM-lactate, there was little change in lactate concentration. 14C from lactate was incorporated into glycogen. There was a marked exponential decrease in lactate specific radioactivity, much greater with [3H]- than with [14C]-lactate. The 'apparent turnover' of [U-14C]lactate was 0.28 mumol/min per g of muscle, and those of [2-3H]- and [3-3H]-lactate were both about 0.7 mumol/min per g. With 10 mM-lactate as sole substrate, there was a net uptake of lactate, at a rate of about 0.15 mumol/min per g, and the apparent turnover of [U-14C]lactate was 0.3 mumol/min per g. The apparent turnover of [3H]lactate was 3-5 times greater. When glycogen synthesis was low (no prestimulation, no insulin), the incorporation of lactate carbon into glycogen exceeded that from glucose, but at high rates of glycogen deposition the incorporation of lactate carbon was much less than that of glucose. Lactate incorporation into glycogen was similar in fast-twitch white and fast-twitch red muscle, but was very low in slow-twitch red fibres. We find that (a) pyruvate in muscle is incorporated into glycogen without randomization of carbon, and synthesis is not inhibited by mercaptopicolinate or cycloserine; (b) there is extensive lactate turnover in the absence of net lactate uptake, and there is a large dilution of 14C-labelled lactate from endogenous supply; (c) there is extensive detritiation of [2-3H]- and [3-3H]-lactate in excess of 14C utilization.     

Glucose Metabolism in Rat Retinal Pigment Epithelium

The retinal pigment epithelium (RPE) is the major transport pathway for exchange of metabolites and ions between choroidal blood supply and the neural retina. To gain insight into the mechanisms controlling glucose metabolism in RPE and its possible relationship to retinopathy, we studied the influence of different glucose concentrations on glycogen and lactate levels and CO₂ production in RPE from normal and streptozotocin-treated diabetic rats. Incubation of normal RPE in the absence of glucose caused a decrease in lactate production and glycogen content. In normal RPE, increasing glucose concentrations from 5.6 mM to 30 mM caused a four-fold increase in glucose accumulation and CO₂ yield, as well as reduction in lactate and glycogen production. In RPE from diabetic rats glucose accumulation did not increase in the presence of high glucose substrate, but it showed a four- and a seven-fold increase in CO₂ production through the mitochondrial and pentose phosphate pathways, respectively. We found high glycogen levels in RPE which can be used as an energy reserve for RPE itself and/or neural retina. Findings further show that the RPE possesses a high oxidative capacity. The large increase in glucose shunting to the pentose phosphate pathway in diabetic retina exposed to high glucose suggests a need for reducing capacity, consistent with increased oxidative stress.     

Developmental Changes in Brain Glucose, Glycogen, Phosphocreatine, and ATP Levels in DBA/2J and C57BL/6J Mice, and Audiogenic Seizures

Abstract. Brains of rodents are primarily dependent on ketone bodies as a source of hydrogen for NADH and of acetyl-CoA during the perinatal period characterized by suckling. A mouse dam begins to wean her pups at about 14 days of age (DOA), the same age at which the brain reaches near-adult size and begins to shift to dependence on carbohydrate-derived sources of acetyl-CoA for normal function. Also at this time, the ear canals open, and mice of some strains become susceptible to audiogenic seizures (AGS). There may be a genetically determined derangement in the orderly transition from one source of brain energy to the other in AGS-prone mice, with a concomitant brief (days) reduction in the in situ energy reserve during the transition. In mice with a decreased energy reserve, a large energy expenditure within a short period of time (s), such as that induced by a substantial acoustic stimulus to newly opened acoustic pathways, might briefly lead to CNS disorganization before body energy repletion processes may occur, resulting in the onset of an AGS. Since glucose, glycogen, ATP, and phosphocreatine provide the bulk of the brain energy reserve, a developmental study was performed to measure the concentrations of these metabolites in brain tissues of DBA/2J mice (genetically/developmentally susceptible to AGS: onset at 12–14 DOA, peak at 18–21 DOA, rapid decline until 30 DOA, essentially lost by 42 DOA) and in C57BL/6J mice (not developmentally susceptible to AGS). Samples of frontal, temporal, cerebellar, and diencephalic regions were taken from mice 0–44 DOA and assayed. With the exception of higher glycogen levels in both DBA and C57 mice in cerebellar and diencephalic samples 0–16 DOA, no regional differences were found. A decrease in glycogen in all regions was observed in DBA mice 16–30 DOA, which was the inverse of susceptibility to AGS in these mice. This dip was not found in C57 mice. ATP levels were elevated in DBA mice 14–18 DOA, and glucose levels were decreased in DBA mice 24–40 DOA. These data lend support to the hypothesis that lowered brain energy reserves, or lowered access to brain reserves, underlies susceptibility to AGS.     

Cardiac metabolism in mice: tracer method developments and in vivo application revealing profound metabolic inflexibility in diabetes

Studies of cardiac fuel metabolism in mice have been almost exclusively conducted ex vivo. The major aim of this study was to assess in vivo plasma FFA and glucose utilization by the hearts of healthy control (db/+) and diabetic (db/db) mice, based on cardiac uptake of (R)-2-[9,10-(3)H]bromopalmitate ([3H]R-BrP) and 2-deoxy-D-[U-14C]glucose tracers. To obtain quantitative information about the evaluation of cardiac FFA utilization with [3H]R-BrP, simultaneous comparisons of [3H]R-BrP and [14C]palmitate ([14C]P) uptake were first made in isolated perfused working hearts from db/+ mice. It was found that [3H]R-BrP uptake was closely correlated with [14C]P oxidation (r2 = 0.94, P < 0.001). Then, methods for in vivo application of [3H]R-BrP and [14C]2-DG previously developed for application in the rat were specially adapted for use in the mouse. The method yields indexes of cardiac FFA utilization (R(f)*) and clearance (K(f)*), as well as glucose utilization (R(g)'). Finally, in the main part of the study, the ability of the heart to switch between FFA and glucose fuels (metabolic flexibility) was investigated by studying anesthetized, 8-h-fasted control and db/db mice in either the basal state or during glucose infusion. In control mice, glucose infusion raised plasma levels of glucose and insulin, raised R(g)' (+58%), and lowered plasma FFA level (-48%), K(f)* (-45%), and R(f)* (-70%). This apparent reciprocal regulation of glucose and FFA utilization by control hearts illustrates metabolic flexibility for substrate use. By contrast, in the db/db mice, glucose infusion raised glucose levels with no apparent influence on cardiac FFA or glucose utilization. In conclusion, tracer methodology for assessing in vivo tissue-specific plasma FFA and glucose utilization has been adapted for use in mice and reveals a profound loss of metabolic flexibility in the diabetic db/db heart, suggesting a fixed level of FFA oxidation in fasted and glucose-infused states.     

Temperature effects on ACTH-stimulated adrenocortical secretion and carbohydrate metabolism in the lizard (Dipsosaurus dorsalis)

Summary Desert iguanas (Dipsosaurus dorsalis) were maintained at 10, 20, 30 or 40°C for one week. Animals were autopsied at zero time, and 30 and 90 minutes after a single injection of ACTH. Basal plasma corticosterone (B) level was similar in all lizards regardless of environmental temperature. Both blood glucose and liver glycogen levels showed highly significant correlations with environmental temperature. Injection of ACTH increased plasma B level in all temperature groups but the rate and magnitude of the response were temperature dependent. Liver glycogen was increased after ACTH injection at 20°C and plasma glucose response to ACTH was significantly correlated with temperature.     

Metabolic effects of testosterone during prolonged physical exercise and fasting

Previous studies have shown a decrease in plasma testosterone during prolonged physical exercise and 72 h fasting in rats. To determine whether this hormonal change has an influence upon energy metabolism, two experiments were carried out, in which the plasma levels of testosterone were elevated during prolonged physical exercise and fasting in male wistar rats. The effects of acute and chronic increases in the levels of circulating testosterone were studied, on the one hand after human chorionic gonadotropin (H.C.G.) injection, and on the other by prolonged testosterone perfusion with an osmotic minipump. Blood and tissue sampling were performed to evaluate blood glucose, alanine, and lactate, and tissue glycogen. The results in fed and rest control rats showed no changes in blood parameters under the effect of hypertestosteronemia but there was an increase in muscle glycogen after testosterone perfusion. In 72 h fasted rats both types of hypertestosteronemia were associated with a decrease in blood alanine and lactate ranging from 25% to 35%. Only testosterone perfusion was associated with higher concentrations of muscle glycogen. After 7 h of treadmill running, testosterone perfusion and H.C.G. injection induced a 35% decrease in blood alanine and a slight decrease in blood glucose, with no change in other parameters. Whereas an elevation in the level of testosterone can induce muscle glycogen compensation in the fed resting state, it cannot counteract the exhaustion of muscle glycogen during running.