Effect of low glycogen on glycogen synthase in human muscle during and after exercise.

Subjects cycled at a work load calculated to elicit 75% of maximal oxygen uptake on two occasions: the first to fatigue (34.5 +/- 5.3 min; mean +/- SE), and the second at the same workload and for the same duration as the first. Biopsies were obtained from the quadriceps femoris muscle before and immediately after exercise, and 5 min post-exercise. Before the first experiment, muscle glycogen was lowered by a combination of exercise and diet, and before the second, experiment muscle glycogen was elevated. In the low glycogen condition (LG), muscle glycogen decreased from 169 +/- 15 mmol glucosyl units kg-1 dry wt at to rest to 13 +/- 6 after exercise. In the high glycogen condition (HG) glycogen decreased from 706 +/- 52 at rest to 405 +/- 68 after exercise. Glycogen synthase fractional activity (GSF) was always higher during the LG treatment. During exercise in the HG condition, those subjects who cycled for less than 35 min (n = 3) had GSF values in muscle which were lower than at rest, whereas those subjects who cycled for greater than 35 min (n = 4) had values which were similar to or higher than at rest. Thus the change in GSF in muscle during HG was positively related to the exercise duration (r = 0.94; y = 254-17x + 0.3x2; P less than 0.001) and negatively related to the glycogen content at the end of exercise (r = -0.82; y = 516-2x + 0.001x2; P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)     

Uptake and degradation of glycogen by Kupffer cells.

Rat liver glycogen was isotopically labeled with [¹⁴C] glucose and isolated. The isolated glycogen was injected intravenously into a series of rats and its vascular clearance, uptake and degradation in liver was analyzed by means of labeling and ultrastructural techniques. Injected glycogen was quickly removed from serum with a $\text{t}\text{1}\text{2}$ of less than 15 min. Glycogen particles, identified in the electron microscope, were never seen to attach to the surface of Kupffer cells or hepatocytes. These particles appeared to be taken up by Kupffer cells by nonspecific pinocytosis “fluid endocytosis” e.g., as a solute with engulfed liquid. By 10 min the particles were present within single membrane bound vacuoles of Kupffer cells. At this early time point, the vacuoles did not seem to have fused with preexisting prelabeled secondary lysosomes containing ferritin. At later time points, glycogen particles were seen intermingled with ferritin. By 1, 2, and 4 hr, increasing numbers of vacuoles containing granular material believed to represent glycogen were observed. These vacuoles often showed extensive enlargement (“swelling”). By 24 hr, most glycogen particles had disappeared and granular material was prresent only in occasional lysosomes which no longer appeared swollen. The estimated half-life for glycogen in Kupffer cell lysosomes was in the range of 12 to 16 hr. This is considerably longer than for membrane proteins and lipids introduced into Kupffer cell lysosomes by means of heterophagy. Because of possible reutilization of isotope it was difficult to define the half-life of glycogen more exactly. It is concluded that glycogen is degraded in Kupffer cell lysosomes, although at a relative slow rate, in comparison with the capability of lysosomal hydrolases to digest proteins and lipids. This conclusion is in line with the general notion that glycogen degradation takes place in the cell sap and is not primarily associated with any particular organelle.     

Distribution of glycogen in prefusion human palatal epithelium.

Various stages of embryonic human secondary palatal development were examined for the presence of epithelial glycogen. Utilizing periodic acid-Schiff's reagent staining of thick plastic sections and osmium ferrocyanide enhancement of thin sections, dramatic changes in epithelial glycogen distribution were noted during palatogenesis. Prior to fusion, the epithelium destined to adhere in the midline exhibited a marked diminution of glycogen in the superficial cell layer. This cell layer was composed of slender dense cells and cuboidal cells undergoing lysis. Adjacent nonfusing epithelium was markedly different and contained large glycogen reserves in its superficial cell layer. Glycogen may play a role either as precursor for specific adhesive macromolecules or as a physical agent capable under the influence of appropriate enzymes of causing cell lysis.     

Muscle glycogen does not interfere with a 13CO2 breath test to monitor liver glycogen oxidation.

Naturally 13C-enriched carbohydrate has been used to label the liver glycogen pool for metabolic studies. The utilization of this glycogen was then monitored by the appearance of 13CO2 in breath. Using this method, it is assumed that during sedentary fasting the contribution of muscle glycogen towards oxidation is negligible. We investigated the influence of a different level of 13C enrichment of muscle glycogen on the 13C enrichment of breath CO2 while the breath test was carried out. In six healthy volunteers, the muscle glycogen stores were grossly depleted by a cycling exercise prior to consumption of the 13C-enriched diet which was given over a 10 h period. The oxidation of liver glycogen was measured during an 18 h sedentary fast. The results were compared with a control group who had not depleted their muscle glycogen before labelling. A higher 13C enrichment of muscle glycogen did not interfere with two parameters of liver glycogen oxidation, i.e. the duration of the plateau phase of 13CO2 and the return to baseline time. It was also shown that the 13C-labelled muscle glycogen was still available after the 18 h fast because a strenuous exercise led to a rapid 13CO2 enrichment. It is concluded that muscle glycogen 13C enrichment does not invalidate a 13CO2 breath test to measure liver glycogen oxidation during a sedentary fast.     

Control of glycogen metabolism in the developing turkey poult.

An experiment was conducted with young turkey poults to evaluate factors controlling glycogen metabolism in the period following hatching. Glucose and sucrose solutions were given along with a standard starter diet. Liver and carcass glycogen were measured on days 1, 4 and 6. Liver glycogen synthetase (EC 2.4.1.21) and phosphorylase (EC 2.4.1.1) were also assayed at these times. The characteristics of active and inactive glycogen synthetase at these times were determined and sensitivity of the active and inactive forms were related to physiological concentrations of glucose-6-phosphate. Supplemental glucose or sucrose increased carcass glycogen in comparison to controls; however, but sucrose was more effective than glucose in promoting liver glycogen synthesis in 4- and 6-day-old poults. There was an age dependent increase in carcass glycogen between days 1 and 6, but a decrease in liver glycogen between days 4 and 6. The activation of liver glycogen synthetase is incomplete in 1 day old poults but activity increases during the 1st week of life. Activation of glycogen synthetase decreased the apparent Ka for glucose-6-phosphate. Phosphorylase inactivation in vitro was not affected by age. Liver glucose-6-phosphate increases rapidly after hatching and the concentration is related to the in vitro Ka derived for both active and inactive synthetases. Both glucose and sucrose increased liver glucose-6-phosphate at days 4 and 6 as well as glycogen synthetase activity. The increase in enzyme activity may be caused indirectly by an allosteric effect of glucose-6-phosphate. Phosphorylase, while not affected by supplemental carbohydrates, did decrease in activity between days 4 and 6. The decrease in activity could affect the phosphorylase a/ synthetase a ratio and change glycogen metabolism.     

High-intensity exercise and muscle glycogen availability in humans.

This study investigated the effects of muscle glycogen availability on performance and selected physiological and metabolic responses during high-intensity intermittent exercise. Seven male subjects completed a regimen of exercise and dietary intake (48 h) to either lower and keep low (LOW-CHO) or lower and then increase (HIGH-CHO) muscle glycogen stores, on two separate occasions at least a week apart. On each occasion the subjects completed a short-term (<10 min) and prolonged (>30 min) intermittent exercise (IEX) protocol, 24 h apart, which consisted of 6-s bouts of high-intensity exercise performed at 30-s intervals on a cycle ergometer. Glycogen concentration (mean +/- SEM) in m. vastus lateralis before both IEx(short) and IEx(long) was significantly lower following LOW-CHO [180 (14), 181 (17) mmol kg (dw)(-1)] compared with HIGH-CHO [397 (35), 540 (25) mmol kg (dw)(-1)]. In both IEx(short) and IEx(long), significantly less work was performed following LOW-CHO compared with HIGH-CHO. In IEx(long), the number of exercise bouts that could be completed at a pre-determined target exercise intensity increased by 265% from 111 (14) following LOW-CHO to 294 (29) following HIGH-CHO (P < 0.05). At the point of fatigue in IEx(long), glycogen concentration was significantly lower with the LOW-CHO compared with HIGH-CHO [58 (25) vs. 181 (46) mmol kg (dw)(-1), respectively]. The plasma concentrations of adrenaline and nor-adrenaline (in IEx(short) and IEx(long)), and FFAand glycerol (in IEx(long)), increased several-fold above resting values with both experimental conditions. Oxygen uptake during the exercise periods in IEx(long), approached 70% of Vo2max. These results suggest that muscle glycogen availability can affect performance during both short-term and more prolonged high-intensity intermittent exercise and that with repeated exercise periods as short as 6 s, there can be a relatively high aerobic contribution.     

Effects of supranormal liver glycogen content on hyperglucagonemia-induced liver glycogen breakdown

The purpose of the present study was to test the hypothesis that a higher hepatic glycogen level is associated with higher glucagon-induced hepatic glycogen depletion. Four groups of anesthetized rats received three injections (at times 0, 30, and 60 min) of glucagon (intravenously, 20 [microg/kg). Among these groups, hepatic glycogen levels had previously been manipulated either by an overloading diet (Fast-refed), a reduction in food intake (1/2-fast), or exercise (75 min of running, 26 m/ min, 0% grade). A fourth group had normal hepatic glycogen levels. A fifth group of rats was injected only with saline (0.9% NaCl). Liver glycogen concentrations were measured every 30 min during the course of the 90-min experiment, using liver samples obtained from the open liver biopsy technique. Plasma glucagon concentrations were significantly higher (P < 0.05) in the glucagon-injected groups than in the saline-injected group. As expected, liver glycogen levels were significantly higher (P < 0.01; 1.6-fold) in the Fast-refed group than in all other groups. Glucagon-induced decreases in liver glycogen concentrations were similar in Fast-refed than in normally fed and exercised rats when the overall 90-min period was considered. However, during the course of the last 30-min period, liver glycogen was significantly (P < 0.01) decreased only in the Fast-refed group. The Fast-refed, normally fed, and exercised groups had a similar glucagon-induced hyperglycemia that was significantly more elevated (P < 0.01) than glucose levels measured in the saline-injected group. Glucagon-induced reactive hyperinsulinemia was observed only in the Fast-refed and normally fed rats, and not in the exercised and 1/2-fast rats. It is concluded that supranormal levels of liver glycogen may be associated with a larger hyperglucagonemia-induced liver glycogen breakdown.     

Regulation of glycogen breakdown by glycogen level in contracting rat muscle

The interaction of glycogen concentration, insulin and beta-adrenergic stimulation in the regulation of glycogen breakdown was studied in perfused rat muscles. Rats were pre-conditioned to obtain two groups with either normal (N) or 'supercompensated' (SC) muscle glycogen. The next day their hindlimbs were perfused with a medium containing insulin (0, 40 and 100 microU mL(-1)) and/or isoproterenol (0 and 1.5 nmol L(-1)). Contractions were induced by electrical stimulation of the sciatic nerve. Compared with N, glycogen breakdown in white gastrocnemius during contractions was greater in SC at any hormonal combination (P < 0.05). Conversely, in red gastrocnemius (RG) the higher glycogenolytic rate in SC, compared with N, faded as the insulin concentration was raised from 0 to 100 microU mL(-1). However, isoproterenol restored the higher glycogenolytic rate in SC. In any condition, RG glycogen synthase fractional activity was lower (P < 0.05) during contractions in SC than in N. Furthermore, the percentage of phosphorylase a was higher in SC except when muscles were exposed to insulin alone. In conclusion, high initial glycogen concentration in fast-glycolytic muscle causes high glycogenolytic rate during contractions, irrespective of hormonal stimulation. In contrast, due to down-regulation of phosphorylase activity, such a relationship does not exist in insulin-stimulated fast-oxidative muscle.     

Effects of supranormal liver glycogen content on hyperglucagonemia-induced liver glycogen breakdown

The purpose of the present study was to test the hypothesis that a higher hepatic glycogen level is associated with higher glucagon-induced hepatic glycogen depletion. Four groups of anesthetized rats received three injections (at times 0, 30, and 60 min) of glucagon (intravenously, 20?μg/kg). Among these groups, hepatic glycogen levels had previously been manipulated either by an overloading diet (Fast-refed), a reduction in food intake (1/2-fast), or exercise (75?min of running, 26?m/min, 0% grade). A fourth group had normal hepatic glycogen levels. A fifth group of rats was injected only with saline (0.9% NaCl). Liver glycogen concentrations were measured every 30?min during the course of the 90-min experiment, using liver samples obtained from the open liver biopsy technique. Plasma glucagon concentrations were significantly higher (P?<?0.05) in the glucagon-injected groups than in the saline-injected group. As expected, liver glycogen levels were significantly higher (P?<?0.01; 1.6-fold) in the Fast-refed group than in all other groups. Glucagon-induced decreases in liver glycogen concentrations were similar in Fast-refed than in normally fed and exercised rats when the overall 90-min period was considered. However, during the course of the last 30-min period, liver glycogen was significantly (P?<?0.01) decreased only in the Fast-refed group. The Fast-refed, normally fed, and exercised groups had a similar glucagon-induced hyperglycemia that was significantly more elevated (P?<?0.01) than glucose levels measured in the saline-injected group. Glucagon-induced reactive hyperinsulinemia was observed only in the Fast-refed and normally fed rats, and not in the exercised and 1/2-fast rats. It is concluded that supranormal levels of liver glycogen may be associated with a larger hyperglucagonemia-induced liver glycogen breakdown.     

肝糖原累积症-I型的肾脏改变

本文观察了14例临床或病理诊断的肝糖原累积症-Ⅰ型(GSD),年龄3月-19岁,5例(36%)伴有肾脏改变.其中1例,8岁,仅血β_2-M轻度升高,4例大于12岁.4例中2例表现为轻度蛋白尿,3例内生肌酐清除率降低,3例β_2-M轻度升高,1例伴继发性Fanconi综合征和肾性佝倭病表现.1例(14岁)进行了肾活检,光镜下示系膜基质轻度增生,肾小管胞浆内有糖原物质沉积.观察结果表明GSD-Ⅰ型病人的肾脏改变较为普遍,应密切追随肾脏情况,以期经过合理饮食治疗后能延缓本病晚期肾功能恶化.     

Polymorphic markers of the glycogen debranching enzyme gene allowing linkage analysis in families with glycogen storage disease type III.

Glycogen storage disease type III (GSD-III), an autosomal recessive disease, is caused by deficient glycogen debranching enzyme (GDE) activity. We identified three polymorphic markers in the GDE gene using single strand conformation polymorphism (SSCP) analysis and DNA sequencing. They were -10G/A in the 5' non-translated region of exon 3,2001 + 8C/T in intron 16, and 3199C/T (P1067S) in exon 25. Two polymorphic markers (-10G/A and 2001 + 8C/T) were highly informative in both controls and GSD-III patients with heterozygosity values of 0.50 and 0.46, respectively. The third marker (3199C/T) had a heterozygosity value of 0.26. Restriction analysis of the PCR amplified genomic DNA products in two GSD-III families showed for the first time the potential use of these markers for carrier detection and prenatal diagnosis in this disease.     

Regulation of glycogen resynthesis in muscles of rats following exercise.

Following a strenuous bout of exercise, glycogen repletion occurred most rapidly in the fast-twitch red type of muscle, least rapidly in fast-twitch white, and at an intermediate rate in slow-twitch red muscle. There was a linear correlation between glycogen synthase I activity and the rate of glycogen synthesis in the three types of muscle. This finding helps explain the differences between the rates of glycogen resynthesis in the three muscle types, and supports the view that glycogen synthase activity is the most important factor determining the rate of glycogen synthesis when substrate supply is adequate. There was an inverse correlation between muscle glycogen concentration and percent glycogen synthase I. Plasma insulin concentration was low and norepinephrine and glucagon concentrations were elevated in the postexercise period. The finding that rapid glycogen synthesis occurred despite a hormonal milieu conducive to glycogenolysis provides evidence that a low glycogen concentration is a potent stimulus to glycogen synthesis that overrides the effects of low insulin, and high norepinephrine and glucagon levels.     

New perspectives on the storage and organization of muscle glycogen.

Due to its large mass, skeletal muscle contains the largest depot of stored carbohydrate in the body in the form of muscle glycogen. Readily visualized by the electron microscope, glycogen granules appear as bead-like structures localized to specific subcellular locales. Each glycogen granule is a functional unit, not only containing carbohydrate, but also enzymes and other proteins needed for its metabolism. These proteins are not static, but rather associate and dissociate depending on the carbohydrate balance in the muscle. This review examines glycogen-associated proteins, their interactions, and roles in regulating glycogen metabolism. While certain enzymes such as glycogen synthase and glycogen phosphorylase have been extensively studied, other proteins such as the glycogen initiating and targeting proteins are just beginning to be understood. Two metabolically distinct forms of glycogen, pro- and marcoglycogen have been identified that vary in their carbohydrate complement per molecule and have different sensitivities to glycogen synthesis and degradation. Glycogen regulation takes place not only by allosteric regulation of enzymes, but also due to other factors such as subcellular location, granule size, and association with various glycogen-related proteins.