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Comparison between starvation and consumption of a high protein diet: plasma insulin and glucagon and hepatic activities of gluconeogenic enzymes during the first 24 hours.

Plasma insulin, glucagon, glucose, free fatty acids and glycerol, hepatic cyclic AMP and glycogen, and liver phosphoenolpyruvate carboxykinase (PEPCK), fructose 1,6-bisphosphatase (FBPase), glucose 6-phosphatase (G6Pase) and alanine amino transferase (AAT) activities were examined in adult rats during the first 24 h of either starvation or consumption of a high protein, carbohydrate-free (HP) diet. Under both nutritional conditions, plasma insulin fell within 12 h and remained constant thereafter. Glucagon increased 12 h after the start of the experiment and peaked between 18-24 h. The insulin: glucagon ratio was lower during the last 12 h of the experiment. In both experimental groups, liver cyclic AMP increased progressively and peaked between 15-24 h, but it increase was higher on HP diet than on starvation. Whereas plasma glucose remained low on starvation for 24 h, it returned to normal on consumption of the HP diet. In both groups, liver glycogen fell within 12 h and remained low until the end of experiment. FBPase, G6Pase and AAT did not change on starvation, while they increased toward the end of 1 d HP consumption. During starvation or consumption of the HP diet, PEPCK increased progressively and peaked between 15-24 h, but the increase was greater with the HP diet than with starvation. These findings suggest that in the first 24 hours, the adaptative response of hepatic gluconeogenesis is higher with a HP diet than upon starvation.

Adaptation, Physiological↗

Structures of the amylopectins of waxy, normal, amylose-extender, and wx:ae genotypes and of the phytoglycogen of maize.

Average chain lengths and beta-amylolysis limits have been determined for the waxy and ae/wx genotypes of mature maize starch, and for the amylopectin fractions of normal and amylose-extender starches (prepared by precipitation with concanavalin A), rabbit-liver glycogen, phytoglycogen, and waxy rice starch. All amylopectin samples had similar A:B chain ratios of > 1 and the two glycogens had ratios of < 1. This finding led to average frequencies of substitution of B chains over the whole molecule of > 2 for the amylopectins and < 2 for the glycogens. An equation for the number of tiers for a molecule with various frequencies of substitution of B chains and chain lengths has been used to determine the effect of variation in average frequency of branching and average chain length on structure.

Amylopectin↗

Stimulation of mitochondrial functions by glucagon treatment, starvation and by treatment of isolated mitochondria with glycogen-bound enzymes.

Liver mitochondria isolated from rats starved overnight, or fed rats injected with glucagon, exhibited a similar increase of the respiration rate with succinate (by 30-40%) and glutamate plus malate (by 20-30%), as compared to mitochondria from control fed animals. The content of mitochondrial adenine nucleotides was elevated by 30-45% by glucagon treatment or starvation. Mitochondrial respiration and citrulline synthesis were stimulated by 30-40% when mitochondria isolated from fed rats were briefly preincubated with the extract from liver glycogen granules, ATP and MgCl2. This effect was abolished by heating the extract at 100 degrees C.

Adenine Nucleotides↗

Synthase activation is not a prerequisite for glycogen synthesis in the starved liver.

To evaluate the contribution of phosphorylase and synthase interconversion as well as the availability of substrates to the onset of liver glycogen synthesis, this process was studied in rats starved overnight and refed for 4 h. On feeding, phosphorylase kinase and phosphorylase were inactivated in a cAMP-independent way, but the proportion of synthase a was unchanged and associated with increased hexoses 6-phosphate (glucose and fructose 6-phosphate), uridine diphosphoglucose (UDPG), and fructose 2,6-bisphosphate concentrations. These findings serve to support a "push" mechanism whereby substrate availability for synthase a concerted with phosphorylase inactivation provokes glycogen deposition. Anesthesia was compulsory for liver sampling and analysis. If such experiments were carried out in conscious rats killed by decapitation, artefactual cAMP-dependent phosphorylase activation and synthase inactivation were observed in starved animals. The phosphorylase activation persisted in refed animals but by a cAMP-independent mechanism.

Animals↗

[Change in tissue glycogen reserves in tumor bearers as a reflection of a hypoglycemic stress syndrome].

Glycogen content in the brain, liver and skeletal muscles of rats bearing ascite Zajdela hepatoma (AZH) and solid 27 hepatoma (27-H) has been studied. Serum glucose levels directly correlated with liver glycogen reserves. In the terminal stage of tumor growth depletion of liver glycogen was observed, while the stores of muscle glycogen did not diminish. Within 1-4 days (AZH) and 15-30 days (27-H) after implantation the stores even exceeded those of control healthy rats. In the terminal stage, in spite of hypoglycaemia development, the content of brain glycogen was significantly elevated in both groups of animals.

Animals↗

Hormonal effects on gluconeogenesis from (U-14C)glutamate in rainbow trout (Salmo gairdneri).

Blood glucose was significantly decreased by insulin (4 I.U./kg). Glucagon (1 mg/kg) and Cortisol (5 mg/kg) administration produced a significant hyperglycaemia. Insulin administration did not modify liver glycogen levels. Glucagon showed a marked liver glycogen mobilization. Cortisol stimulated liver glycogen deposition. Insulin and Glucagon showed a significant inverse effect on gluconeogenesis from (U-14C)glutamate, decreasing and increasing 14C-glucose formation respectively. Hormonal treatments did not influence the very low levels of incorporation of (U-14C)glutamate into liver and muscle glycogen.

Animals↗

High-affinity binding of glycogen-synthase phosphatase to glycogen particles in the liver. Role of glycogen in the inhibition of synthase phosphatase by phosphorylase a.

1. Post-mitochondrial supernatants were prepared from the livers of 24 h-fasted rats. Upon centrifugation at high speed, the major part of the glycogen-synthase phosphatase activity sedimented with the microsomal fraction. However, two approaches showed that the enzyme was associated with residual glycogen rather than with vesicles of the endoplasmic reticulum. Indeed, the activity was entirely solubilized when the remaining glycogen was degraded either by glucagon treatment in vivo or by alpha-amylolysis in vitro. No evidence could be found for an association of glycogen-synthase phosphatase with the smooth endoplasmic reticulum, as isolated with the use of discontinuous sucrose gradients. 2. After solubilization by glucagon treatment in vivo, synthase phosphatase could be transferred to glycogen particles with very high affinity. Half-maximal binding occurred at a glycogen concentration of about 0.25 mg/ml, whereas glycogen synthase and phosphorylase required 1.5-2 mg/ml. 3. In gel-filtered extracts prepared from glycogen-depleted livers, the activation of glycogen synthase was not inhibited at all by phosphorylase alpha. The inhibition was restored when the liver homogenates were prepared in a glycogen-containing buffer. The effect was half-maximal at a glycogen concentration of about 0.25 mg/ml, and virtually complete at 1 mg/ml. These findings explain long-standing observations that in fasted animals the liver contains appreciable amounts of both synthase and phosphorylase in the active form.

Animals↗

Preferential resynthesis of muscle glycogen in fasting rats after exhausting exercise.

Despite carbohydrate starvation (fasting or fat feeding) considerable glycogen accumulation, ranging from 20 to 30 mumol glucose/g, occurred in hindlimb muscles of rats following exhausting exercise that caused severe muscle and liver glycogen depletion and hypoglycemia. The largest increase in muscle glycogen occurred during the first 3 h after exercise when plasma levels of glucagon and epinephrine were very high and insulin concentration was low. The concentrations of glycogen attained in different hindlimb muscles in the fasting and fat-fed animals were between 50 and 100% of the values found in rats fed carbohydrate after the exhausting exercise. In rats fed carbohydrate following exercise, liver glycogen accumulation greatly exceeds muscle glycogen accumulation. A remarkable difference in the response of liver glycogen was seen in the carbohydrate starved rats. In contrast to the rapid increase in muscle glycogen, liver glycogen was still essentially completely depleted in the fasting and fat-fed rats 24 h after exercise. This indicates that the glucose made available via gluconeogenesis was preferentialy channeled away from liver glycogen synthesis into muscle glycogen.

Animals↗

Effect of swimming in thermal water on skeletal muscle, liver and heart glycogen.

Skeletal muscle, liver and heart glycogen variations, induced by swimming in thermal water (at 35 degrees C) as a model of physical exercise for clinical use, were studied. Muscle and liver glycogen moderately decreases after a 30-min period of swimming and comes near to depletion after 60 min. Heart glycogen decreases only slightly after 60 min. Blood glucose and plasma insulin decrease only after 60 min of swimming. A 30-min swim in thermal water, cooled to 25 degrees C, depletes muscle and liver glycogen and slightly decreases heart glycogen. Under these conditions, plasma insulin decreases and hypoglycemia occurs. The results seem to indicate some advantages of swimming in hot thermal water in order to prevent glycogen store depletion as the physiological prerequisite for a physical exercise of clinical interest to obtain therapeutical benefits, avoiding premature fatigue and exhaustion.

Animals↗

The effect of temperature on immunoreactive glucagon plasma level in carp Cyprinus carpio.

Plasma immunoreactive glucagon levels (IRG), plasma glucose levels and brain and liver glycogen concentrations were analyzed in carp (adapted to 15 degrees C) subjected to short-term temperature changes (1.6 or 11 h, at 5 degrees C or 28 degrees C, respectively) and to long-term temperature changes (21 months at 28 degrees C). The high temperature (28 degrees C) produced significant increases in IRG in both short and long-term experiments. Brain glycogen also decreased in both experiments whereas liver glycogen only changed in the long-term experiment. Low temperatures did not provoke any changes either in IRG or in liver glycogen, whereas brain glycogen decreased in the 1 h exposure. In short, under these conditions in carp, IRG did not respond to low temperature but could play an important role in high temperature acclimation.

Animals↗

Glycogen metabolism as detected by in vivo and in vitro 13C-NMR spectroscopy using [1,2-13C2]glucose as substrate.

The metabolism of glucose to glycogen in the liver of fasted and well-fed rats was investigated with 13C nuclear magnetic resonance spectroscopy using [1,2-(13)C2]glucose as the main substrate. The unique spectroscopic feature of this molecule is the 13C-13C homonuclear coupling leading to characteristic doublets for the C-1 and C-2 resonances of glucose and its breakdown products as long as the two 13C nuclei remain bonded together. The doublet resonances of [1,2-(13)C2]glucose thus provide an ideal marker to follow the fate of this exogenous substrate through the metabolic pathways. [1,2-(13)C2]Glucose was injected intraperitoneally into anesthetized rats and the in vivo 13C-NMR measurements of the intact animals revealed the transformation of the injected glucose into liver glycogen. Glycogen was extracted from the liver and high resolution 13C-NMR spectra were obtained before and after hydrolysis of glycogen. Intact [1,2-13C2]glucose molecules give rise to doublet resonances, natural abundance [13C]glucose molecules produce singlet resonances. From an analysis of the doublet-to-singlet intensities the following conclusions were derived. (i) In fasted rats virtually 100% of the glycosyl units in glycogen were 13C-NMR visible. In contrast, the 13C-NMR visibility of glycogen decreased to 30-40% in well-fed rats. (ii) In fed rats a minimum of 67 +/- 7% of the exogenous [1,2-(13)C2]glucose was incorporated into the liver glycogen via the direct pathway. No contribution of the indirect pathway could be detected. (iii) In fasted rats externally supplied glucose appeared to be consumed in different metabolic processes and less [1,2-(13)C2]glucose was found to be incorporated into glycogen (13 +/- 1%). However, the observation of [5,6-(13)C2]glucose in liver glycogen provided evidence for the operation of the so-called indirect pathway of glycogen synthesis. The activity of the indirect pathway was at least 9% but not more than 30% of the direct pathway. (vi) The pentose phosphate pathway was of little significance for glucose but became detectable upon injection of [1-(13)C]ribose.

Animals↗

The glycogen-depleting effect of exogenous maternal epinephrine on the liver of the fetal rat during the sixteenth day of gestation.

The effect of 1-epinephrine administered I. P. to pregnant rats during sodium pentobarbital anesthesia on fetal liver glycogen levels has been investigated. Sodium pentobarbital (22 mg./kg. maternal body weight) was administered I. P. to all pregnant rats in the control group and each 1-epinephrine dosage group twenty minutes prior to the surgical removal of the fetuses. L-epinephrine (10, 20 or 30 micrograms/kg. maternal body weight) given ten minutes after the administration of the barbiturate reduced the fetal liver glycogen levels significantly. The glycogen-depleting effect of 1-epinephrine on the fetal liver in the rat was found to be dosage-dependent.

Animals↗

Hyperglycemic effect of neurotensin, a hypothalamic peptide.

A hypotensive, gut-contracting peptide neurotensin (NT), recently isolated from bovine hypothalami, has been found to produce hyperglycemia within minutes after iv injection into anesthetized rats. The dose-response relationship (deltaglucose, 15 min after injection) was linear over the range 30-200 pmol/100 g BW. NT did not alter the disappearance rate of [14C]glucose from plasma during the development of the hyperglycemia. However, the peptide caused a fall in liver glycogen (52 +/- 6.5 to 41 +/- 3.3 mg/g) and a 7-fold increase in the activity of the 5'-AMP independent form of liver glycogen phosphorylase. Activation of liver glycogen phosphorylase did not occur in vitro under conditions found suitable for demonstrating the effectiveness of glucagon, suggesting the possible involvement of an intermediary substanc(s) in vivo. Acute adrenalectomy did not prevent the response. Hypophysectomized rats (4 days post-operative) were less sensitive to NT, perhaps as a consequence of their diminished liver glycogen levels (normal, 52 +/- 6.5 mg/g; hypophysectomized, 23 +/- 1.8 mg/g); however, the presence of the pituitary was not essential for this response. NT was also effective in rats with hereditary diabetes insipidus (Brattleboro strain). At the time intervals sampled, radioimmunoassayable plasma levels of growth hormone, glucagon, and insulin were not significantly changed after injection of NT into normal rats. Pretreatment of rats with reserpine (7 mg/kg), morphine sulfate (10 mg/kg), propranolol (5mg/kg), or phenoxybenzamine (10 mg/kg) did not prevent the response. These findings characterize the action of NT on liver glycogen metabolism and blood glucose levels, but a physiological role for NT in this regard remains to be demonstrated.

Adrenal Glands↗

Glycogen and glycogen-hydrolysing lysosomal enzyme activity in mouse liver: effects of fasting, adrenoceptor antagonism and insulin-induced hypoglycaemia.

Whereas the phosphorolytic breakdown of liver glycogen is known to be of great physiological importance, the functional role of the hydrolytic glycogenolysis in the lysosomal system is less well understood. In the present study the effects of fasting, alpha- and beta-adrenoceptor antagonism and insulin-induced hypoglycaemia on liver lysosomal glycogen-hydrolysing enzyme activity were investigated in mice. In freely fed mice the glycogen-hydrolysing activity (acid amyloglucosidase) was only 50% of the maltose-hydrolysing activity (acid maltase). Starvation for 24 h reduced the acid amyloglucosidase activity by approximately 30% (P less than 0.001), whereas the activities of acid maltase, acid phosphatase and beta-glucuronidase appeared unaffected. N-acetyl-beta-D-glucosaminidase activity was moderately (20%; P less than 0.01) enhanced by fasting. Thus, liver lysosomal enzyme activities may change independently of each other during fasting. Further, during short-term hypoglycaemic conditions (45 min) induced by endogenous or exogenous insulin, the activity of liver acid amyloglucosidase was found to be moderately reduced (15-20%). Blockade of alpha- and beta-adrenoceptors by phentolamine and propranolol did not result in any apparent influence on acid amyloglucosidase activity except for the indirect effect exerted by the phentolamine-induced hypoglycaemia. A moderate negative correlation (r = -0.51; P less than 0.001) between total liver glycogen concentration and acid amyloglucosidase activity was observed in a series of 43 freely fed NMRI mice. Our data show that in mouse liver the acid maltase activity predominates over the acid amyloglucosidase activity.(ABSTRACT TRUNCATED AT 250 WORDS)

Adrenergic alpha-Antagonists↗