Glycogen Phosphorylase Research Articles

Structure activity relationship of the binding of p-coumaroyl glucose to glycogen phosphorylase and its effect on hepatic cell metabolic pathways

The binding of p-coumaroyl glucose to glycogen phosphorylase (GP; a pharmaceutical target for the development of antihyperglycaemic drugs) has been studied by kinetics, and X-ray crystallography while its effect to HepG2 cells metabolism has been assessed by NMR metabolomics. p-Coumaroyl glucose is a potent inhibitor of human liver GP with a Ki value of 213 μΜ that binds at the active site of the enzyme. Comparative structural analysis with chemically similar GP inhibitors reveals the structural basis of its inhibitory potency. NMR metabolomics analysis revealed that HepG2 cells in the presence of p-coumaroyl glucose actively response to higher glucose uptake from their environment and a display an “insulin-sensitizing’’ state. Furthermore, NMR metabolomics analysis indicates an enhancement of gluconeogenesis towards lipid metabolism and glycerol-derived components.

Effects of tea polyphenols on the growth performance, carbohydrate metabolism of grass carp ( Ctenopharyngodon idellus )

To investigate the effects of tea polyphenols (TPs) on the growth and carbohydrate metabolism of grass carp (Ctenopharyngodon idella), 0 (control group), 250, 500, 1000, 2000, and 4000 mg kg−1 TPs were added to the basal diet and fed to grass carps for 8 weeks. The results show that 500 mg kg−1 group exhibited the significantly highest feed conversion efficiency (FCE) and specific growth rate (SGR) (p < .05) and significantly higher activities of pyruvate kinase (PK), lactate dehydrogenase (LDH), malate dehydrogenase (MDH) and succinate dehydrogenase (SDH) (p < .05). The livers of the control group and 500 mg kg−1 group were subjected to RNA-Seq, and the expressions of glycogen phosphorylase (PYG) and sucrase-isomaltase (SI) genes were upregulated, while that of glucose-6-phosphatase (G6PC) was downregulated, and these promoted glycogen catabolism. The expressions of glucokinase (GK), PK, 6-phosphofructokinase (PFK), glucose-6-phosphate 1-dehydrogenase (G6PD) and fructose-bisphosphate aldolase A (ALDOA) genes were upregulated to promote glucose catabolism. The results indicate that 500 mg kg−1 TPs could upregulate the expressions of some genes that promote carbohydrate decomposition, increase the activities of carbohydrate decomposition enzymes and enhance the utilization of carbohydrates, thereby improving the FCE and SGR.

Human hair follicles operate an internal Cori cycle and modulate their growth via glycogen phosphorylase

Hair follicles (HFs) are unique, multi-compartment, mini-organs that cycle through phases of active hair growth and pigmentation (anagen), apoptosis-driven regression (catagen) and relative quiescence (telogen). Anagen HFs have high demands for energy and biosynthesis precursors mainly fulfilled by aerobic glycolysis. Histochemistry reports the outer root sheath (ORS) contains high levels of glycogen. To investigate a functional role for glycogen in the HF we quantified glycogen by Periodic-Acid Schiff (PAS) histomorphometry and colorimetric quantitative assay showing ORS of anagen VI HFs contained high levels of glycogen that decreased in catagen. qPCR and immunofluorescence microscopy showed the ORS expressed all enzymes for glycogen synthesis and metabolism. Using human ORS keratinocytes (ORS-KC) and ex vivo human HF organ culture we showed active glycogen metabolism by nutrient starvation and use of a specific glycogen phosphorylase (PYGL) inhibitor. Glycogen in ORS-KC was significantly increased by incubation with lactate demonstrating a functional Cori cycle. Inhibition of PYGL significantly stimulated the ex vivo growth of HFs and delayed onset of catagen. This study defines translationally relevant and therapeutically targetable new features of HF metabolism showing that human scalp HFs operate an internal Cori cycle, synthesize glycogen in the presence of lactate and modulate their growth via PYGL activity.

Hepatic glycogenolysis is determined by maternal high-calorie diet via methylation of Pygl and it is modified by oteocalcin administration in mice

ObjectiveAccumulating evidence indicates that an adverse perinatal environment contributes to a higher risk of metabolic disorders in the later life of the offspring. However, the underlying molecular mechanisms remain largely unknown. Thus, we investigated the contribution of maternal high-calorie diet and osteocalcin to metabolic homeostasis in the offspring. MethodsEight-week-old C57Bl/6N female mice were mated with age-matched males and allocated randomly to three groups: a normal-diet (ND) or a high-fat, high-sucrose diet group, which was administered either saline (control) or GluOC (10 ng/g body mass) from the day of mating to that of delivery, and the dams were fed a ND after the delivery. Pups weaned at 24 days after birth were analyzed. ResultsA maternal high-fat, high-sucrose diet during pregnancy causes metabolic disorders in the liver of the offspring via hypermethylation of the Pygl gene, encoding glycogen phosphorylase L, which mediates hepatic glycogenolysis. The reduced expression of Pygl induced by the maternal diet causes the hepatic accumulation of glycogen and triglyceride in the offspring, which remains in adulthood. In addition, the administration of uncarboxylated osteocalcin during pregnancy upregulates Pygl expression via both direct CREBH and ATF4 and indirect epigenomic pathways, mitigating the maternal diet-induced obesity and abnormal glucose and lipid metabolism in adulthood. ConclusionsWe propose that maternal energy status is reflected in the hepatic glycogenolysis capacity of the offspring via epigenetic modification of Pygl and uncarboxylated osteocalcin regulates glycogenolysis.

Astrocytic A1/A2 paradigm participates in glycogen mobilization mediated neuroprotection on reperfusion injury after ischemic stroke

BackgroundAstrocytic glycogen works as an essential energy reserve for surrounding neurons and is reported to accumulate excessively during cerebral ischemia/reperfusion (I/R) injury. Our previous study found that accumulated glycogen mobilization exhibits a neuroprotective effect against I/R damage. In addition, ischemia could transform astrocytes into A1-like (toxic) and A2-like (protective) subtypes. However, the underlying mechanism behind accumulated glycogen mobilization-mediated neuroprotection in cerebral reperfusion injury and its relationship with the astrocytic A1/A2 paradigm is unknown.MethodsAstrocytic glycogen phosphorylase, the rate-limiting enzyme in glycogen mobilization, was specifically overexpressed and knocked down in mice and in cultured astrocytes. The I/R injury was imitated using a middle cerebral artery occlusion/reperfusion model in mice and an oxygen–glucose deprivation/reoxygenation model in cultured cells. Alterations in A1-like and A2-like astrocytes and the expression of phosphorylated nuclear transcription factor-κB (NF-κB) and phosphorylated signal transducer and activator of transcription 3 (STAT3) were determined by RNA sequencing, immunofluorescence and immunoblotting. Metabolites, including glycogen, NADPH, glutathione and reactive oxygen species (ROS), were analyzed by biochemical analysis.ResultsHere, we observed that astrocytic glycogen mobilization inhibited A1-like astrocytes and enhanced A2-like astrocytes after reperfusion in an experimental ischemic stroke model in vivo and in vitro. In addition, glycogen mobilization could enhance the production of NADPH and glutathione by the pentose phosphate pathway (PPP) and reduce ROS levels during reperfusion. NF-κB inhibition and STAT3 activation caused by a decrease in ROS levels were responsible for glycogen mobilization-induced A1-like and A2-like astrocyte transformation after I/R. The astrocytic A1/A2 paradigm is closely correlated with glycogen mobilization-mediated neuroprotection in cerebral reperfusion injury.ConclusionsOur data suggest that ROS-mediated NF-κB inhibition and STAT3 activation are the key pathways for glycogen mobilization-induced neuroprotection and provide a promising metabolic target for brain reperfusion injury in ischemic stroke.

Inhibition of RAS components attenuates progression of heart failure and its adverse consequences on myocardial remodeling in both normotensive and hypertensive rats

Abstract Rationale and purpose Prevention or attenuation of heart failure (HF) is persistent task in clinic as well as challenge for progress in research. Both, intercellular connexin-43 (Cx43) channels and extracellular matrix (ECM) exert high impact on heart function. Thus, we aimed to explore whether HF due to volume overload (VO) induces alterations in myocardial Cx43, ECM proteins MMP2, SMAD2/3, TGFβ1 and PKC signaling as well as changes that may be revealed by enzyme histochemistry. Moreover, we tested the impact of treatment suppressing RAS on examined targeted proteins. Methods VO-HF was induced in male normotensive Hannover Sprague- Dawley (HSD) and mRen-2 transgenic hypertensive (TGR) rats by creating an aorto-caval fistula (ACF) and heart response was examined 20-weeks later. Sham-rats were compared with non-treated rats with ACF and those treated for 15-weeks with ACEI (trandolapril, 6mg/l, p.o.) or ARB (losartan (200mg/l, p.o.). Left ventricular heart (LV) tissue was analyzed using western blot, zymography and enzyme histochemistry. Key results Echocardiography confirmed an increase in cardiac output and a decrease in ejection fraction in both strains of rats. BW was higher in TGR vs HSD and it was not affected by VO. HW and LVW were higher in TGR vs HSD and VO increased both parameters regardless the strain while ACEI&amp;gt;ARB attenuated it. In parallel, myocardial pro-hypertrophic PKCd expression was higher in TGR vs HSD and increased in both strains due to VO while both drugs prevent it. Glycogen phosphorylase and capillary associated 5-nucleotidase, alkaline phospatase and dipeptidyl peptidase IV activities were reduced due to VO in HSD and TGR and it was attenuated by treatment. Cx43 expression and its functional phosphorylation status were lower in TGR vs HSD and suppressed in both strains due to VO but ACEI&amp;gt;ARB prevented it. Similar trend of changes was observed in PKCe expression, which phosphorylates Cx43. Profibrotic TGFβ1-Smad2/3 pathway was suppressed along with reduced MMP2 activity in TGR comparing to HSD. VO reduced SMAD2/3 in HSD and TGFβ1 in TGR as well as MMP2 expression in both strains, whereby treatment abolished these changes. VO decreased MMP2 (63kDa) activity only in TGR and ARB prevented it. VO did not affect collagen deposition in either strain vs sham rats. Conclusions Hypertrophic and hypofibrotic phenotypes are induced by volume overload in both rat strains. Together with deterioration of Cx43 mediated intercellular coupling it may contribute to heart dysfunction. Inhibition of RAS components attenuates progression of VO and its adverse consequences. Funding Acknowledgement Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Scientific Grant Agency of the Ministry of Education, Science, Research and Sport of the Slovak Republic VEGA 2/0158/19, 2/0002/20

Glycogen accumulation and phase separation drives liver tumor initiation

Glucose consumption is generally increased in tumor cells to support tumor growth. Interestingly, we report that glycogen accumulation is a key initiating oncogenic event during liver malignant transformation. We found that glucose-6-phosphatase (G6PC) catalyzing the last step of glycogenolysis is frequently downregulated to augment glucose storage in pre-malignant cells. Accumulated glycogen undergoes liquid-liquid phase separation, which results in the assembly of the Laforin-Mst1/2 complex and consequently sequesters Hippo kinases Mst1/2 in glycogen liquid droplets to relieve their inhibition on Yap. Moreover, G6PC or another glycogenolysis enzyme-liver glycogen phosphorylase (PYGL) deficiency in both human and mice results in glycogen storage disease along with liver enlargement and tumorigenesis in a Yap-dependent manner. Consistently, elimination of glycogen accumulation abrogates liver growth and cancer incidence, whereas increasing glycogen storage accelerates tumorigenesis. Thus, we concluded that cancer-initiating cells adapt a glycogen storing mode, which blocks Hippo signaling through glycogen phase separation to augment tumor incidence.

Molecular Functions and Pathways of Plastidial Starch Phosphorylase (PHO1) in Starch Metabolism: Current and Future Perspectives.

Starch phosphorylase is a member of the GT35-glycogen-phosphorylase superfamily. Glycogen phosphorylases have been researched in animals thoroughly when compared to plants. Genetic evidence signifies the integral role of plastidial starch phosphorylase (PHO1) in starch biosynthesis in model plants. The counterpart of PHO1 is PHO2, which specifically resides in cytosol and is reported to lack L80 peptide in the middle region of proteins as seen in animal and maltodextrin forms of phosphorylases. The function of this extra peptide varies among species and ranges from the substrate of proteasomes to modulate the degradation of PHO1 in Solanum tuberosum to a non-significant effect on biochemical activity in Oryza sativa and Hordeum vulgare. Various regulatory functions, e.g., phosphorylation, protein–protein interactions, and redox modulation, have been reported to affect the starch phosphorylase functions in higher plants. This review outlines the current findings on the regulation of starch phosphorylase genes and proteins with their possible role in the starch biosynthesis pathway. We highlight the gaps in present studies and elaborate on the molecular mechanisms of phosphorylase in starch metabolism. Moreover, we explore the possible role of PHO1 in crop improvement.

Association of milk production and udder type traits with polymorphism of phosphorylase kinase regulatory subunit alpha-2 gene in Sahiwal cattle

The study was conducted in Sahiwal cattle in tropical region in the Indian sub-continent where the production of dairy animals is yet to be optimised. The present investigation was executed to identify SNPs in PHKA2 gene and to explore its effect on udder type and milk production traits. The study was based on the hypothesis that the PHKA2 gene has highly variable exons that could be related with udder traits and eventually milk production. PHKA2 gene regulates glycogen phosphorylase a, a catalyst in breakdown of glycogen. Milk production traits were recorded; 9 udder type, 5 teat type and 8 visual traits were measured for 100 animals. Five highly variable targeted regions of PHKA2 gene were amplified using PCR and sequenced. The association analysis was carried out using general linear model (SAS) to study the fixed effect of genotype on studied traits. The synonymous type SNP g.124556852C&gt;T was found for the first time in Sahiwal cattle and possibly associated with udder type traits. The genotypic frequencies with respect to targeted loci g.124556852C&gt;T indicated that homozygote CC (0.58) were highest in our resource population. The chi-square 2 test showed an agreement to Hardy–Weinberg equilibrium. The association analysis revealed significant association of genotypes with udder width, udder balance and 305 days milk yield. The attempt to find significant association with the visual udder traits was also done, however no significant alliance was observed. Homozygote CC animal were desirable as they favoured the selection of animal with superior udder width, udder balance and 305 day’s Milk Yield values.

Chebulagic acid attenuates HFD/streptozotocin induced impaired glucose metabolism and insulin resistance via up regulations of PPAR γ and GLUT 4 in type 2 diabetic rats

The aim of the present study was to evaluate the effects of chebulagic acid on enzymes involved in carbohydrate metabolism in high-fat diet/streptozotocin-induced diabetic rats. Diabetes was induced in albino male Wistar rats by feeding them with a high-fat diet comprising of 84.3% standard laboratory rat chow, 5% lard, 10% yolk powder, 0.2% cholesterol, and 5% bile salt for 2 weeks. After 2 weeks, the animals were kept in an overnight fast and injected with a low dose of streptozotocin (35 mg/kg, dissolved in 0.1 M sodium citrate buffer, pH 4.5). At the end of the experimental periods, the blood glucose, plasma insulin, hemoglobin, glycated hemoglobin levels, homeostatic model of insulin resistance, glycogen content, glycogen synthase, glycogen phosphorylase were measured in high-fat diet and streptozotocin-induced diabetic rats whereas the glucose metabolic enzymes such as hexokinase, glucose 6 phosphate dehydrogenase, glucose 6 phosphatase, fructose 1,6 bisphosphatase and protein expression of peroxisome proliferator-activated receptor–γ (PPAR-γ) and Glucose transporter 4 (GLUT4) were analyzed. Administration of chebulagic acid to HFD/STZ -induced diabetic rats for 30 days significantly decreased the levels of plasma glucose, homeostatic model assessment of insulin resistance, glycogen phosphorylase and glycosylated hemoglobin and increased the levels of insulin, glycogen content and glycogen synthase. On the other hand, the altered activity of carbohydrate metabolic enzymes, PPAR-γ and GLUT4 were brought back to near-normal levels on treatment with chebulagic acid. The effect produced by chebulagic acid on various parameters was comparable to that of metformin.

Two Adaptations of Foray's Unified Energetic Budget Measurement Protocol for Highly Chitinized Insects and Faster Glycogen Determination

Two Adaptations of Foray's Unified Energetic Budget Measurement Protocol for Highly Chitinized Insects and Faster Glycogen Determination

Vanillin modulates activities linked to dysmetabolism in psoas muscle of diabetic rats

Skeletal muscles are important in glucose metabolism and are affected in type 2 diabetes (T2D) and its complications. This study investigated the effect of vanillin on redox imbalance, cholinergic and purinergic dysfunction, and glucose-lipid dysmetabolism in muscles of rats with T2D. Male albino rats (Sprague–Dawley strain) were fed 10% fructose ad libitum for 2 weeks before intraperitoneally injecting them with 40 mg/kg streptozotocin to induce T2D. Low (150 mg/kg bodyweight (BW)) and high (300 mg/kg BW) doses of vanillin were orally administered to diabetic rats. Untreated diabetic rats and normal rats made up the diabetic control (DC) and normal control (NC) groups, respectively. The standard antidiabetic drug was metformin. The rats were humanely put to sleep after 5 weeks of treatment and their psoas muscles were harvested. There was suppression in the levels of glutathione, activities of SOD, catalase, ENTPDase, 5′Nucleotidase and glycogen levels on T2D induction. This was accompanied by concomitantly elevated levels of malondialdehyde, serum creatine kinase-MB, nitric oxide, acetylcholinesterase, ATPase, amylase, lipase, glucose-6-phosphatase (G6Pase), fructose-1,6-biphophastase (FBPase) and glycogen phosphorylase activities. T2D induction further resulted in the inactivation of fatty acid biosynthesis, glycerolipid metabolism, fatty acid elongation in mitochondria and fatty acid metabolism pathways. There were close to normal and significant reversals in these activities and levels, with concomitant reactivation of the deactivated pathways following treatment with vanillin, which compared favorably with the standard drug (metformin). Vanillin also significantly increased muscle glucose uptake ex vivo. The results suggest the therapeutic effect of vanillin against muscle dysmetabolism in T2D as portrayed by its ability to mitigate redox imbalance, inflammation, cholinergic and purinergic dysfunctions, while modulating glucose-lipid metabolic switch and maintaining muscle histology.

Exercise-Induced Lactate Release Mediates Mitochondrial Biogenesis in the Hippocampus of Mice via Monocarboxylate Transporters.

Regular exercise training induces mitochondrial biogenesis in the brain via activation of peroxisome proliferator-activated receptor gamma-coactivator 1α (PGC-1α). However, it remains unclear whether a single bout of exercise would increase mitochondrial biogenesis in the brain. Therefore, we first investigated whether mitochondrial biogenesis in the hippocampus is affected by a single bout of exercise in mice. A single bout of high-intensity exercise, but not low- or moderate-intensity, increased hippocampal PGC-1α mRNA and mitochondrial DNA (mtDNA) copy number at 12 and 48h. These results depended on exercise intensity, and blood lactate levels observed immediately after exercise. As lactate induces mitochondrial biogenesis in the brain, we examined the effects of acute lactate administration on blood and hippocampal extracellular lactate concentration by in vivo microdialysis. Intraperitoneal (I.P.) lactate injection increased hippocampal extracellular lactate concentration to the same as blood lactate level, promoting PGC-1α mRNA expression in the hippocampus. However, this was suppressed by administering UK5099, a lactate transporter inhibitor, before lactate injection. I.P. UK5099 administration did not affect running performance and blood lactate concentration immediately after exercise but attenuated exercise-induced hippocampal PGC-1α mRNA and mtDNA copy number. In addition, hippocampal monocarboxylate transporters (MCT)1, MCT2, and brain-derived neurotrophic factor (BDNF) mRNA expression, except MCT4, also increased after high-intensity exercise, which was abolished by UK5099 administration. Further, injection of 1,4-dideoxy-1,4-imino-D-arabinitol (glycogen phosphorylase inhibitor) into the hippocampus before high-intensity exercise suppressed glycogen consumption during exercise, but hippocampal lactate, PGC-1α, MCT1, and MCT2 mRNA concentrations were not altered after exercise. These results indicate that the increased blood lactate released from skeletal muscle may induce hippocampal mitochondrial biogenesis and BDNF expression by inducing MCT expression in mice, especially during short-term high-intensity exercise. Thus, a single bout of exercise above the lactate threshold could provide an effective strategy for increasing mitochondrial biogenesis in the hippocampus.

Diarylheptanoid‐flavanone Hybrids as Multiple‐target Antidiabetic Agents from Alpinia katsumadai

Main observation and conclusionThe EtOAc fraction of Alpinia katsumadai seeds showed significant inhibition on glycogen phosphorylase a (GPa) with inhibitory ratios of 97.9% and 64.6% at concentrations of 200 and 100 μg/mL, respectively. Bioactivity‐guided isolation afforded 15 new diarylheptanoid‐flavanone hybrids, katsumadainols B1—B15 (1—15), together with eight known ones (16—23). Compounds 4—10 and 12—21 exhibited activity against GPa with IC50 values of 10.1—95.4 μmol/L; compounds 4, 5, 16, and 17 displayed inhibitory effects on α‐glucosidase with IC50 values of 7.1, 12.4, 7.2, and 8.3 μmol/L, obviously higher than acarbose (IC50, 209.1 μmol/L); compounds 4—6, 14, 16—20, 22, and 23 were PTP1B/TCPTP selective inhibitors with IC50 values of 40.7—95.8 μmol/L; compounds 4, 5, 16, and 17 showed DPP4 inhibitory effects with inhibitory ratios of 50.0%—54.2% (200 μmol/L). Diarylheptanoid‐flavanone hybrids (4, 5, 16, and 17) with a p‐hydroxybenzyl at C‐6 position represent a promising class of multiple‐target antidiabetic agents inhibiting GPa, α‐glucosidase, PTP1B, and DPP4.

Galangin Attenuates Liver Injury, Oxidative Stress and Inflammation, and Upregulates Nrf2/HO-1 Signaling in Streptozotocin-Induced Diabetic Rats

Chronic hyperglycemia increases the risk of liver damage. Oxidative stress and aberrant inflammatory response are entangled in diabetes-associated liver injury. This study evaluated the protective effect of the flavonoid galangin (Gal) on glucose intolerance, liver injury, oxidative stress, inflammatory response, and Nrf2/HO-1 signaling in diabetic rats. Diabetes was induced by streptozotocin (STZ), and the rats received Gal for six weeks. STZ-induced rats showed glucose intolerance, hypoinsulinemia, elevated glycated hemoglobin (HbA1c), and decreased liver glycogen. Gal ameliorated glucose intolerance, reduced HbA1c%, increased serum insulin and liver glycogen and hexokinase activity, and suppressed glycogen phosphorylase, glucose-6-phosphatase and fructose-1,6-biphosphatase in diabetic rats. Circulating transaminases, ALP and LDH, and liver ROS, MDA, TNF-α, IL-1β, and IL-6 were increased and GSH, SOD, and CAT were diminished in diabetic rats. In addition, diabetic rats exhibited multiple histopathological alterations and marked collagen deposition. Treatment with Gal mitigated liver injury, prevented histopathological alterations, decreased ROS, MDA, pro-inflammatory cytokines, Bax and caspase-3, and enhanced cellular antioxidants and Bcl-2. Gal downregulated hepatic Keap1 in diabetic rats and upregulated Nrf2 and HO-1 mRNA as well as HO-1 activity. Molecular modeling studies revealed the ability of Gal to bind to and inhibit NF-κB and Keap1, and also showed its binding pattern with HO-1. In conclusion, Gal ameliorates hyperglycemia, glucose intolerance, oxidative stress, inflammation, and apoptosis in diabetic rats. Gal improved carbohydrate metabolizing enzymes and upregulated Nrf2/HO-1 signaling.

Glycogen phosphorylase revisited: extending the resolution of the R- and T-state structures of the free enzyme and in complex with allosteric activators.

The crystal structures of free T-state and R-state glycogen phosphorylase (GP) and of R-state GP in complex with the allosteric activators IMP and AMP are reported at improved resolution. GP is a validated pharmaceutical target for the development of antihyperglycaemic agents, and the reported structures may have a significant impact on structure-based drug-design efforts. Comparisons with previously reported structures at lower resolution reveal the detailed conformation of important structural features in the allosteric transition of GP from the T-state to the R-state. The conformation of the N-terminal segment (residues 7-17), the position of which was not located in previous T-state structures, was revealed to form an α-helix (now termed α0). The conformation of this segment (which contains Ser14, phosphorylation of which leads to the activation of GP) is significantly different between the T-state and the R-state, pointing in opposite directions. In the T-state it is packed between helices α4 and α16 (residues 104-115 and 497-508, respectively), while in the R-state it is packed against helix α1 (residues 22'-38') and towards the loop connecting helices α4' and α5' of the neighbouring subunit. The allosteric binding site where AMP and IMP bind is formed by the ordering of a loop (residues 313-326) which is disordered in the free structure, and adopts a conformation dictated mainly by the type of nucleotide that binds at this site.

Endometrial glycogen metabolism on days 1 and 11 of the reproductive cycle in dairy cows

Embryos need glucose or fructose to remain viable; however, it is not well understood how secretion of these carbohydrates is regulated. This study was conducted to evaluate endometrial glycogen and associated enzymes on Days 1 and 11 of the estrous cycle (Day 0 = behavioral estrus) in cattle. Diastase-liable periodic acid–Schiff (PAS) staining of luminal epithelia decreased 81 % between Days 1 and 11. Similarly, glycogen content of glandular epithelia was 66 % less on Day 11 than Day 1. There was dense PAS staining in the lumen of some glands, and this staining was removed when there was pretreatment with diastase. Based on western blot results, there was no difference in glycogen metabolizing enzymes between Days 1 and 11. Results from conducting immunohistochemistry procedures indicated hexokinase 1 was more abundant in the epithelial cells than stroma, but immunostaining was not different between Day 1 and 11. In contrast, phospho-glycogen synthase was undetectable on Day 1 but was present in glandular epithelia on Day 11. Glycogen synthase was localized to the epithelia, and was in larger abundance on Day 1. The abundance of glycogen phosphorylase was greater in the epithelium than stroma and on Day 11 than 1. Furthermore, glucose-6-phosphatase 3 was more abundant in the epithelium on both Days 1 and 11. In conclusion, in the uterus of cattle glycogen is stored in a reproductive cycle-dependent manner. Glucose released from endometrial glycogen stores could potentially be utilized by the endometrium or secreted into the uterine lumen.

In vitro fertilization causes excessive glycogen accumulation in mouse placenta

IntroductionChildren conceived by assisted reproductive technologies have a high risk of suffering from obstetrical complications and long-term health problems, but the related mechanisms are not fully understood. Normal placental function is closely linked with foetal growth and future health. Given the significance of glycogen metabolism in placentas, we investigated the effect of in vitro fertilization (IVF) on glycogen storage in placentas using a mouse model. MethodsMouse placentas were collected at E18.5 after natural mating or IVF, and the placental and foetal weights were recorded. The quantitative assay kit and histological staining were used to measure the glycogen content. Additionally, we detected the expression of multiple genes associated with glycogen synthesis/decomposition, glucose transporters, and the phosphorylation of Akt and Gsk3β. ResultsOur findings showed that IVF resulted in a significantly increased mouse placental weight and enlarged junctional area. We found, compared to the control, excessive glycogen was accumulated in IVF placentas. However, we observed that multiple genes involved in glycogen generation (Gsk3b, Phka1, Phkb, Phkg1, and Phkg2) and glycogenolysis (Agl and Pygm) had lower mRNA levels in IVF placentas. Moreover, the expression levels of glycogen synthase, phosphorylase, Glut1, and Glut3 were significantly decreased in IVF placentas. The phosphorylation activities of Akt Ser473 and Gsk3β Ser9 were inhibited in IVF placentas. DiscussionIVF leads to enlarged mouse placentas with excessive glycogen storage in late pregnancy, and these abnormal changes may be associated with the activation of the Akt-Gsk3β pathway.

Protective Effects of Giant Salamander Bioactive Peptides on D-galactose Induced Oxidative Damage in Mice

To investigate anti-aging mechanism of giant salamander bioactive peptides, label-free quantitative (LFQ) proteomics was used after high-abundant proteins were removed. The results of LFQ showed 425 low-abundant proteins in serum in mice, in which 26 proteins were differentially expressed (P<0.05), and covered 14 biological processes, 6 molecular functions and 7 cellular components. Tetrahydrofolate synthase and glycogen phosphorylase were up-regulated expressed significantly (P<0.05), increased by 1.78 times and 1.05 times respectively, while xanthine dehydrogenase was down-regulated expressed significantly (P<0.05) decreased. The expression of these enzymes increased or decreased, which could reduce the amount of free radicals in vivo. In summary, the giant salamander bioactive peptides prevented aging by scavenging free radicals, this study would provide basic data for the use of the giant salamander bioactive peptides.

An early-postmortem metabolic comparison among three extreme acute heat stress temperature settings in chicken breast muscle.

Normally, preslaughter acute heat stress could accelerate postmortem glycolysis and impair chicken breast (pectoralis major muscle) quality. However, previous studies indicated that it might be different when the acute heat stress temperature rises to an extreme range (above 35°C). Therefore, this study's objectives were to compare the pH decline, glycolytic enzyme activity, and AMP-activated protein kinase (AMPK) phosphorylation at early postmortem among three extreme acute heat stress temperature settings: a control group (36°C) and two experimental groups (38°C and 40°C). Although the temperature did not affect glycogen phosphorylase a and pyruvate kinase activity, there was a decrease in pH decline rate, phosphofructokinase-1 activity, and phospho-AMPK-α[Thr172] within 4h postmortem when temperature increased from 36 to 40°C. Temperature also affected hexokinase activity, with the 36°C-group having the highest activity. The results of the current study, for the first time, indicated that postmortem metabolic rate in chicken breast muscle could be changed by acute heat stress temperature setting at extreme range.