Zonal distribution of glycogen synthesis in isolated rat hepatocytes

Incubation of hepatocytes isolated from fasted rats with [14C]glucose for short periods of time showed that the initial stages of glycogen synthesis occur near the plasma membrane. Incubation with [14C]glucose followed by cold glucose demonstrated that glycogen synthesis is always active at the hepatocyte periphery and that previously synthesised glycogen moves towards the centre of the cell, while its place is filled by newly synthesised molecules. However, the reverse experiment, incubation with cold glucose before addition of [14C]glucose, showed that, as glycogen synthesis progresses, it also becomes gradually active in more internal sites of the hepatocyte. These results indicate a spatial order in the synthesis of hepatic glycogen.     

Dynamics of glycogen content in the normal and cirrhotic liver following glucose administration to starving rats

Using biochemical, cytofluorimetric and television cytophotometric methods, glycogen contents were studied in normal and cirrhotic rat liver at various intervals after glucose administration to fasting animals. The obtained data indicate that after a 48 h fasting glycogen contents in normal and cirrhotic liver are equally poor. A marked rise of glycogen content in cirrhotic liver was observed only 20-30 min after glucose administration to rats. It has been established that at all intervals after glucose administration to rats hepatocytes of the portal lobule zone, both in normal and in cirrhotic liver, accumulate more glycogen than those of the central zone. Again, the intensity of glycogen accumulation in cirrhotically altered liver is significantly lower than in normal liver, due, presumably, to a lower rate of glycogen synthesis in pathologically changed liver.     

Zonal distribution and seasonal vertical migration of copepod assemblages in the Scotia Sea

Large, biomass-dominant Southern Ocean copepod species have been much studied, but small and mesopelagic species also play major rôles in these ecosystems. However, little is known of some basic aspects of their ecology. To address this, the abundances of 23 copepod species and genera were analysed from 72 stations sampled during the Discovery Expeditions in the 1920s to 1950s. Stratified net samples, usually to a depth of 1000?m, provided year-round coverage in the Scotia Sea from the Subantarctic Front to the Weddell-Scotia Confluence. Small copepods (Microcalanus pygmaeus, Ctenocalanus spp., Oncaea spp. and Oithona spp.) formed ～75% of total copepod abundance in the top 1000?m across all major zones. Oithona spp. composed ～40% of copepod numbers in the Polar Front area and to its south: further north their importance declined. All mesopelagic taxa except for the warmer-water species Metridia lucens and Pleuromamma robusta, extended throughout the entire study area, with smaller regional differences than for the shallower-living species. The species showed a continuum of temperature ranges, and there was no evidence that the Polar Front was a major biogeographic boundary to their distribution. Indeed, several important species, including Oithona spp. (mainly Oithona similis), Ctenocalanus spp., Metridia lucens and Rhincalanus gigas reached maximum numbers in this area. Total copepod abundance was thus higher in the vicinity of the Polar Front than in any other region. Only two copepod families made pronounced seasonal vertical migrations: Eucalaniidae (Eucalanus longiceps and R. gigas) and Calaniidae (Neocalanus tonsus, Calanoides acutus, Calanus simillimus and Calanus propinquus). Some evidence for a winter descent was found for Ctenocalanus spp. and some deeper-living groups: Euchaeta spp. and the Metridiidae, although their migrations were not so great as for the eucalanids and calanids.     

Lack of seasonal changes in mitochondrial DNA variability of a Drosophila subobscura population

Restriction site analysis of mtDNA of 550 isofemale lines corresponding to different seasonal samples of a single geographic population of Drosophila subobscura was carried out. The distribution pattern of haplotypes was similar to that observed for the entire range of the species on the European continent: two haplotypes were equally and highly frequent, and a set of sporadic haplotypes were almost never present in more than one seasonal sampling. No statistically significant evidence was found for between-population heterogeneity across time, and the mean within-population variation was similar to other mtDNA restriction site analyses previously reported for D. subobscura populations. These observations could be interpreted in terms of the neutral mutation hypothesis where the entire population has not yet reached an equilibrium. The causes of this non-equilibrium are most likely periodic winter bottlenecks, phenomena that particularly affect the distribution of mtDNA haplotypes across time.     

Different changes of glycogen content in rat muscle denervated by different procedures

Glycogen levels in the extensor digitorum longus of the rat undergo different changes after cutting the sciatic nerve compared with the destruction of the lumbosacral spinal cord. The differences most probably are due to early changes of the endocrine (adrenal and pancreatic) function after the latter type of denervation.     

Physiological and pathological changes in glucose regulate brain Akt and glycogen synthase kinase-3

Insulin regulates the phosphorylation and activities of Akt and glycogen synthase kinase-3 (GSK3) in peripheral tissues, but in the brain it is less clear how this signaling pathway is regulated in vivo and whether it is affected by diabetes. We found that Akt and GSK3 are sensitive to glucose, because fasting decreased and glucose administration increased by severalfold the phosphorylation of Akt and GSK3 in the cerebral cortex and hippocampus of non-diabetic mice. Brain Akt and GSK3 phosphorylation also increased after streptozotocin administration (3 days), which increased blood glucose and depleted blood insulin, indicating regulation by glucose availability even with deficient insulin. Changes in Akt and GSK3 phosphorylation and activities in epididymal fat were opposite to those of brain after streptozotocin treatment. Streptozotocin-induced hyperglycemia and increased brain Akt and GSK3 phosphorylation were reversed by lowering blood glucose with insulin administration. Long term hyperglycemia also increased brain Akt and GSK3 phosphorylation, both 4 weeks after streptozotocin and in db/db insulin-resistant mice. Thus, the Akt-GSK3 signaling pathway is regulated in mouse brain in vivo in response to physiological and pathological changes in insulin and glucose.     

Effect of indomethacin on hepatic glucose production in vitro: the impact of hepatic glycogen content

The effect of indomethacin (IND) on glucagon-induced hepatic glucose production (HGP) was studied in the isolated perfused livers of rats. Addition of IND (0.2 mM) to the perfusion medium had no effect on glucagon-stimulated HGP when compared to control experiments without added IND (1.02 +/- 0.17 vs. 1.00 +/- 0.26 mmol per (120 min X 100 g b.w.), respectively; NS). Intravenous pretreatment with both, IND (10 mg/kg b.w.), or vehicle resulted in a reduction in glucagon-induced HGP due to a decrease in hepatic glycogen content. A complete depletion of the hepatic glycogen pool and thus a lack in glucagon-stimulated HGP was observed when IND was given intraperitoneally. These results indicate that the changes in HGP observed after pretreatment with IND may largely if not completely be due to a non-specific depletion in hepatic glycogen content and that IND does not exert a direct influence on HGP.     

Influence of light regimens on circadian changes in the blood glucose and tissue glycogen concentration in the rat

Adult male Wistar rats allowed food and drinking water ad libitum were kept 2- 5 weeks under standard conditions, but with a different artificial light regimens. The standard regimen was 12:12 h light and dark (LD) and the other variants were 12:12 h dark and light (DL), continuous darkness (DD) and continuous light (LL). The blood glucose level and the liver, skeletal muscle, heart and brown and white adipose tissue glycogen concentration were tested during the day at 3-hour intervals. The experiments were carried out during the winter and were evaluated by an analysis of variance and the cosinor test. Circadian variation of the blood glucose level and the liver and both the adipose tissue glycogen concentrations was only slightly affected by changes in the light regimen. Conversely, the oscillations of the skeletal muscle and heart glycogen concentration reacted markedly to the variants of the light regimen, their reaction being manifested in the localization of the acrophases in different parts of the day, especially when comparing the DD and LD regimens.     

Regulation of glycogen synthesis and glucose utilization in Escherichia coli during maintenance of the energy charge. Quantitative correlation of changes in the rates of glycogen synthesis and glucose utilization with simultaneous changes in the cellular levels of both glucose 6-phosphate and fructose 1,6-diphosphate.

Treatment of nitrogen-starved cultures of Escherichia coli W4597(K) with sodium azide results in simultaneous changes in both glucose 6-phosphate and fructose 1,6-diphosphate as well as in the rate of glycogen synthesis. Based on these observations, a comprehensive equation was developed which relates the cellular levels of both of these hexose phosphates with the rate of glycogen synthesis. This relationship apparently represents the interaction in vivo between the rate-limiting enzyme of bacterial glycogen synthesis, glucose 1-phosphate adenylyltransferase (adenosine diphosphoglucose synthetase, EC 2.7.7.27), and its substrate glucose 1-phosphate (reflected by glucose 6-phosphate) and its major allosteric activator fructose diphosphate. The form of the equation that describes this relationship was determined from studies presented here of the kinetic properties of the E. coli W4597(K) enzyme in the presence of physiological concentrations of its substrates and modulators. We show here and in subsequent reports of this series that the comprehensive relationship between glycogen synthesis and hexose phosphates can serve as a reference to evaluate the possible participation of new factors in the regulation of glycogen synthesis. Treatment with NaN3 did not change the cellular level of glucose 1-phosphate adenylyltransferase. The value of the adenylate energy charge, (ATP + 1/2 ADP)/(ATP + ADP + AMP), was maintained despite losses of up to 35% in cellular adenylates. The quantitative co-variance between hexose phosphates and the cellular rate of glucose utilization that we previously described for other metabolic conditions was also observed in the azide-treated cultures. We integrate the new information into the system of coordinated regulation of glycogen synthesis, glycolysis, and glucose utilization that we proposed previously.     

Seasonal changes in glycogen content and Na+-K+-ATPase activity in the brain of crucian carp

Changes in the number of Na+-K+-ATPase alpha-subunits, Na+-K+-ATPase activity and glycogen content of the crucian carp (Carassius carassius) brain were examined to elucidate relative roles of energy demand and supply in adaptation to seasonal anoxia. Fish were collected monthly around the year from the wild for immediate laboratory assays. Equilibrium dissociation constant and Hill coefficient of [3H]ouabain binding to brain homogenates were 12.87+/-2.86 nM and -1.18+/-0.07 in June and 11.93+/-2.81 nM and -1.17+/-0.06 in February (P>0.05), respectively, suggesting little changes in Na+-K+-ATPase alpha-subunit composition of the brain between summer and winter. The number of [3H]ouabain binding sites and Na-K-ATPase activity varied seasonally (P<0.001) but did not show clear connection to seasonal changes in oxygen content of the fish habitat. Six weeks' exposure of fish to anoxia in the laboratory did not affect Na+-K+-ATPase activity (P>0.05) confirming the anoxia resistance of the carp brain Na pump. Although anoxia did not suppress the Na pump, direct Q10 effect on Na+-K+-ATPase at low temperatures resulted in 10 times lower catalytic activity in winter than in summer. Brain glycogen content showed clear seasonal cycling with the peak value of 203.7+/-16.1 microM/g in February and a 15 times lower minimum (12.9+/-1.2) in July. In winter glycogen stores are 15 times larger and ATP requirements of Na+-K+-ATPase at least 10 times less than in summer. Accordingly, brain glycogen stores are sufficient to fuel brain function for about 8 min in summer and 16 h in winter, meaning about 150-fold extension of brain anoxia tolerance by seasonal changes in energy supply-demand ratio.     

Glucose release in mantle tissue of Mytilus: regulation by calcium ions

Glucose release activity in mantle tissue of Mytilus galloprovincialis was studied. Mantle tissue shows a basal glucose releasing activity. The external Ca2+ absence increases 2 to 3-fold the basal glucose release, and when A23187 (10 microM) was simultaneously present the release doubled that obtained in Ca2(+)-absence. EGTA (2 mM), chlorpromazine (200 microM) and lanthanum (3 mM) decreased the glucose release promoted by external Ca2+ absence. This and other data suggest that glucose release activity in mantle tissue might be controlled by Ca2+ ions.