Inhibition Of Glycogen Synthesis Research Articles

A novel siRNA targeting and delivery platform inhibits glycogen synthesis and reduces glycogen levels in skeletal and cardiac muscle in a mouse model of Pompe disease

A novel siRNA targeting and delivery platform inhibits glycogen synthesis and reduces glycogen levels in skeletal and cardiac muscle in a mouse model of Pompe disease

Nuclear Nicotinamide Adenine Dinucleotide Deficiency by Nmnat1 Deletion Impaired Hepatic Insulin Signaling, Mitochondrial Function, and Hepatokine Expression in Mice Fed a High-Fat Diet

Metabolic syndrome (MetS) is a worldwide challenge that is closely associated with obesity, nonalcoholic liver disease, insulin resistance, and type 2 diabetes. Boosting nicotinamide adenine dinucleotide (NAD+) presents great potential in preventing MetS. However, the function of nuclear NAD+ in the development of MetS remains poorly understood. In this study, hepatocyte-specific Nmnat1 knockout mice were used to determine a possible link between nuclear NAD+ and high-fat diet (HFD)-induced MetS. We found that Nmnat1 knockout significantly reduced hepatic nuclear NAD+ levels but did not exacerbate HFD-induced obesity and hepatic triglycerides accumulation. Interestingly, loss of Nmnat1 caused insulin resistance. Further analysis revealed that Nmnat1 deletion promoted gluconeogenesis but inhibited glycogen synthesis in the liver. Moreover, Nmnat1 deficiency induced mitochondrial dysfunction by decreasing mitochondrial DNA (mtDNA)-encoded complexes Ⅰ and Ⅳ, suppressing mtDNA replication and mtRNA transcription and reducing mtDNA copy number. In addition, Nmnat1 depletion affected the expression of hepatokines in the liver, particularly downregulating the expression of follistatin. These findings highlight the importance of nuclear NAD+ in maintaining insulin sensitivity and provide insights into the mechanisms underlying HFD-induced insulin resistance.

Chronic heat stress inhibits glycogen synthesis through gga-miR-212-5p/GYS1 axis in the breast muscle of broilers

Studies have demonstrated that chronic heat stress can accelerate glycolysis, decrease glycogen content in muscle, and affect muscle quality. However, the consequences of chronic heat stress on glycogen synthesis, miRNA expression in pectoralis major (PM) muscle, and its regulatory functions remain unknown. In this study, high-throughput sequencing and cell experiments were used to explore the effects of chronic heat stress on miRNA expression profiles and the regulatory mechanisms of miRNAs in glycogen synthesis under chronic heat stress. In total, 144 cocks were allocated into 3 groups: the normal control (NC) group, the heat stress (HS) group, and the pair-fed (PF) group. In total, 30 differently expressed (DE) miRNAs were screened after excluding the effect of feed intake, which were mainly related to metabolism, signal transduction, cell growth and death. Furthermore, the gga-miR-212-5p/GYS1 axis was predicted to participate in glycogen synthesis through the miRNA-mRNA analysis, and a dual-luciferase reporter test assay confirmed the target relationship. Mechanistically, chronic heat stress up-regulated gga-miR-212-5p, which could inhibit the expression of GYS1 in the PM muscle. Knocking down gga-miR-212-5p alleviates the reduction of glycogen content caused by chronic heat stress; overexpression of gga-miR-212-5p can reduce glycogen content. This study provided another important mechanism for the decreased glycogen contents within the PM muscle of broilers under heat stress, which might contribute to impaired meat quality.

MicroRNA-92b in the skeletal muscle regulates exercise capacity via modulation of glucose metabolism.

Exercise mimetics is a proposed class of therapeutics that specifically mimics or enhances the therapeutic effects of exercise. Muscle glycogen and lactate extrusion are critical for physical performance. The mechanism by which glycogen and lactate metabolism are manipulated during exercise remains unclear. This study aimed to assess the effect of miR-92b on the upregulation of exercise training-induced physical performance. Adeno-associated virus (AAV)-mediated skeletal muscle miR-92b overexpression in C57BLKS/J mice, and global knockout of miR-92b mice were used to explore the function of miR-92b in glycogen and lactate metabolism in skeletal muscle. AAV-mediated UGP2 or MCT4 knockdown in WT or miR-92 knockout mice was used to confirm whether miR-92b regulates glycogen and lactate metabolism in skeletal muscle through UGP2 and MCT4. Body weight, muscle weight, grip strength, running time and distance to exhaustion, and muscle histology were assessed. The expression levels of muscle mass-related and function-related proteins were analysed by immunoblotting or immunostaining. Global knockout of miR-92b resulted in normal body weight and insulin sensitivity, but higher glycogen content before exercise exhaustion (0.8538±0.0417 vs. 1.043±0.040, **P=0.0087), lower lactate levels after exercise exhaustion (4.133±0.2589 vs. 3.207±0.2511, *P=0.0279), and better exercise capacity (running distance to exhaustion, 3616±86.71 vs. 4231±90.29, ***P=0.0006; running time to exhaustion, 186.8±8.027 vs. 220.8±3.156, **P=0.0028), as compared with those observed in the control mice. Mice skeletal muscle overexpressing miR-92b (both miR-92b-3p and miR-92b-5p) displayed lower glycogen content before exercise exhaustion (0.6318±0.0231 vs. 0.535±0.0194, **P=0.0094), and higher lactate accumulation after exercise exhaustion (4.5±0.2394 vs. 5.467±0.1892, *P=0.01), and poorer exercise capacity (running distance to exhaustion, 4005±81.65 vs. 3228±149.8, ***P<0.0001; running time to exhaustion, 225.5±7.689 vs. 163±6.476, **P=0.001). Mechanistic analysis revealed that miR-92b-3p targets UDP-glucose pyrophosphorylase 2 (UGP2) expression to inhibit glycogen synthesis, while miR-92b-5p represses lactate extrusion by directly target monocarboxylate transporter 4 (MCT4). Knockdown of UGP2 and MCT4 reversed the effects observed in the absence of miR-92b in vivo. This study revealed regulatory pathways, including miR-92b-3p/UGP2/glycogen synthesis and miR-92b-5p/MCT4/lactate extrusion, which could be targeted to control exercise capacity.

Synergistic Effects of Chlorogenic Acid and Ganoderma Lucidum Polysaccharide on Hepatic Fat Accumulation

This study aimed to investigate the impact of different doses of chlorogenic acid and Ganoderma lucidum polysaccharide on lipid accumulation using an obese rat model. We utilized blood glucose assays, oil red O staining, and triglyceride content analysis to examine the effects of chlorogenic acid combined with G. lucidum polysaccharide on fat metabolism in rats. We found substantial potential in using chlorogenic acid combined with G. lucidum polysaccharide for weight control and metabolic regulation in obese rats. After a long-term dietary intervention, chlorogenic acid and G. lucidum polysaccharide markedly inhibited the expression of proteins related to fatty acid synthesis, including AMP-activated protein kinase, fatty acid synthase, and SREBP-1c, leading to reduced body weight and decreased hepatic fat deposition. Additionally, the combined treatment substantially affected enzymes and proteins involved in glucose metabolism, reducing glycogen synthase expression and indicating a potential mechanism for weight reduction through glycogen synthesis inhibition. Furthermore, elevated glucose kinase levels indicated that chlorogenic acid and G. lucidum polysaccharide may enhance hepatic glycogen degradation, thereby reducing hepatic fat accumulation. These findings reveal that chlorogenic acid combined with G. lucidum polysaccharide has a synergistic impact, providing a theoretical basis for their potential as a combined therapeutic approach for obesity.

Impact of particulate matter 2.5 on the liver function of mice.

The aim of this study was to evaluate the impact of particulate matter 2.5 (PM2.5) on liver function at the animal level and to study its impact targets. 60 male and female BALB/c mice of SPF grade, aged 6-8 weeks, were randomly divided into four groups, with 15 mice in each, including the normal saline control group, the PM2.5 low dose group [2 μg/(100 g/d)], the PM2.5 medium dose group [8 μg/(100 g/d)] and the PM2.5 high dose group [16 μg/(100 g/d)]. Each day, 0.9% saline or PM2.5 particles were administered through the nasal route, and samples were taken after 3 weeks of continuous exposure. Hematoxylin-eosin staining (HE) was used to observe the liver damage caused by PM2.5. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were detected by using an automatic biochemical analyzer to detect the content of liver glycogen and blood glucose. Multiple indicators were observed, including plasma tumor necrosis factor (TNF-α) and interleukin-6 (IL-6) levels, oxidative stress response indicators reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD) detection, RT-PCR and Western blot detection of glycogen synthase (GS), glucokinase (GK), nuclear factor erythroid 2-related factor 2 (Nrf2) expression and phosphorylation level of phospho-c-Jun N-terminal kinases (p-JNK). PM2.5 can cause damage to the liver by increasing PM2.5 concentrations, raising the metabolic rate of liver cells, resulting in a substantial amount of inflammatory infiltration and vacuolar degeneration of cells, and increasing the liver/body weight. TNF-α and IL-6 inflammatory factor expression increased (p<0.05). An increase in the serum ALT and AST levels were also observed. The blood glucose of mice increased, whereas the content of liver glycogen declined (p<0.05). ROS, MDA, and SOD levels all increased considerably. PM2.5 can drastically lower the expression of GS and GK, increase the expression of Nrf2, and raise the phosphorylation level of p-JNK (p<0.05). PM2.5 can induce oxidative stress in mouse liver through the Nrf2/JNK pathway, induce liver inflammation in mice, and inhibit glycogen synthesis.

Interruption of bile acid enterohepatic circulation inhibits glycogen synthesis and promotes hepatocellular carcinoma progression

Interruption of bile acid enterohepatic circulation inhibits glycogen synthesis and promotes hepatocellular carcinoma progression

The involvement of GSK3β for glycogen synthesis throughout the annual cycle of Pacific oyster, Crassostrea gigas (Magallana gigas)

Crassostrea gigas is a frequently studied species in understanding physiological processes in bivalves. Similar to other animals, oysters store glucose in the body as glycogen. Glycogen is known to supply energy for germ cell development and maintenance. Glycogen is synthesized by glycogen synthase. GSK3β regulates glycogen synthase activity and plays an important role in glycogen synthesis. Therefore, this study aims to examine the effect of GSK3β on the annual cycle of oysters and the glycogen synthesis pathway and to investigate the energy pathway in comparison with seasonal variation. Oysters were sampled monthly for one year and were subjected to glycogen content, RT-PCR, FISH, and western blot analysis. The year-round glycogen content significantly differs only in the mantle edge during spring and summer of both sexes but not in labial palp, digestive gland, gonad, and adductor muscle. The expression of GSK3β mRNA level was highest in October for females and April for males. Both sexes had the lowest expression in July. In the adductor muscle, females and males showed the highest expression in April and the lowest in July and October. The pattern of GSK3β expression in gonads and adductor muscle was similarly confirmed through FISH. As a result of examining the signaling system, p-GSK3β (serine 9) increased. At the same time, glycogen synthase decreased in May when the condition index was the highest, p-GSK3β decreased in October and July when spawning occurred, and glycogen synthase increased. Overall, it is thought that p-GSK3β expression is high in C. gigas at ripe, which inhibits glycogen synthesis and is used as energy for growth and maturation. Glycogen synthesis occurs for energy storage during degeneration.

The new horizons for treatment of Late-Onset Pompe Disease (LOPD)

Late-onset Pompe disease (LOPD) is a genetic myopathy causing skeletal muscle weakness and severe respiratory impairment, due to the deficiency of the lysosomal enzyme acid alpha-glucosidase (GAA) leading to lysosomal glycogen accumulation along with other complex pathophysiological processes. A major step for treatment of Pompe disease was reached in 2006 with the marketing of alglucosidase alfa, a first enzyme replacement therapy (ERT) that showed a significant motor and respiratory benefit. However, efficacy of alglucosidase alfa is limited in LOPD with a loss of efficacy over time, promoting research on new treatments. Next-generation ERT are new enzymes biochemically modified to increase the uptake of exogenous enzyme by target tissues, and the benefit of two recombinant enzymes (avalglucosidase alfa and cipaglucosidase alfa) has been recently studied in large phase III clinical trials, the latest combined with miglustat. Several innovative therapies, based on GAA gene transfer, antisense oligonucleotides or inhibition of glycogen synthesis with substrate reduction therapy, are currently under study, but are still at an early stage of development. Overall, active research for new treatments raises hope for LOPD patients but challenges remain for the clinician with the need for reliable efficacy assessment tools, long-term registry data, and evidence-based recommendations for the best use of these new molecules recently available or under development.

GLP-1 responds to postprandial hyperglycemia by reducing transcription level in grass carp (Ctenopharyngodon idella)

Glucagon-like peptide-1 (GLP-1), a member of the proglucagon derived peptide family, is a highly effective incretin to reduce plasma glucose and inhibit food intake in mammals. The physiological function of GLP-1 has been elucidated in some ‘glucose intolerant’ carnivorous fishes. To lay a better understanding in fish glucose intolerance, we synthesized homologous GLP-1 peptide and explored its roles in appetite and glucose regulation under intraperitoneal (IP) injection in herbivorous grass carp which has higher glucose intolerance. The results showed that GLP-1 increased plasma glucose in grass carp. The hyperglycemic effect of the fish GLP-1 was mainly through promoting gluconeogenesis, accompanied by inhibition of glycogen synthesis and stimulation of glycogenolysis pathway. In addition, GLP-1 upregulated the expression of anorexigenic gene (cart) and decreased the orexigenic genes (npy and agrp) expression in grass carp hypothalamus as a conservative way to inhibit the appetite under GLP-1 injection after 2 h. Taken together, the hyperglycemic effect and anorectic role of GLP-1 were performed in a basically consistent manner across fishes with different food habits. What’s more, the gene expressions of proglucagon yielded GLP-1 in foregut and hindgut of grass carp were significantly decreased after a meal within 24 h, indicating that fish might suppress the accumulation of plasma glucose by decreasing GLP-1 expression in a unique way to herbivorous fish. Therefore, this study provided a new perspective for illustrating the difference in glucose utilization among fishes with different food habits based on GLP-1.

Roles of adenosine monophosphate-activated protein kinase (AMPK) in the kidney.

Roles of adenosine monophosphate-activated protein kinase (AMPK) in the kidney.

Abstract 3046: In situ analysis of microenvironmental glycogen in Ewing's sarcoma patient samples by mass spectrometry imaging

Abstract Ewing's sarcoma (ES) is a rare pediatric cancer of the bone and soft tissues affecting adolescents and young adults, with peak incidence from ages 10 to 15 years. A clinical feature of ES is the accumulation of Periodic acid-Schiff positive (PAS+) aggregates. PAS+ staining in ES histopathology analysis has been speculated to be glycogen accumulation, however, this has not been confirmed. To interrogate these aggregates, we developed a novel workflow for the high-sensitive detection of in situ glycogen via enzymatic release of glycogen substrates coupled to matrix assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) from formalin-fixed patient tissues. We utilized this method to study the nature of PAS+ staining in a cohort of ES patient samples surgically removed from tibia, rib, chest, shoulder, abdomen and testicle tissues. Using this innovative technique, we discovered profound intra-tumoral glycogen accumulation compared to adjacent stroma, normal, muscle and/or necrotic tissue. Further, intra-tumoral glycogen also has an increased phosphate content and distinct chain length profile compared to normal glycogen found in adjacent and patient-matched tissue. Notably, genetic manipulation and pharmacological inhibition of glycogen synthesis, or removal of glycogen phosphates, blunted ES tumor growth in xenografted mouse models. These findings suggest an important role of ES glycogen aggregates in tumor growth. Together, application of this novel MALDI-IMS technique, and the insights provided by formalin-fixed ES patient samples, reveals a distinct role of ES glycogen aggregates and uncovers a new therapeutic opportunity for this rare pediatric cancer. Citation Format: Lyndsay E. Young, Lindsey R. Conroy, Harrison A. Clarke, Derek B. Allison, Richard R. Drake, Matthew S. Gentry, Ramon C. Sun. In situ analysis of microenvironmental glycogen in Ewing's sarcoma patient samples by mass spectrometry imaging [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 3046.

DhHP-6 ameliorates hepatic oxidative stress and insulin resistance in type 2 diabetes mellitus through the PI3K/AKT and AMPK pathway.

Insulin resistance is one major features of type 2 diabetes mellitus (T2DM). Deuterohemin-βAla-His-Thr-Val-Glu-Lys (DhHP-6), a novel microperoxidase mimetic designed and synthesized based on microperoxidase 11 (MP-11), can scavenge reactive oxygen species (ROS) in vivo. In our previous studies, we showed that oral DhHP-6 could reduce blood glucose and improve insulin resistance. To investigate the mechanisms of how DhHP-6 ameliorates oxidative stress and insulin resistance, we established T2DM mouse models and glucosamine-induced HepG2 cell insulin resistance models. The results suggested that DhHP-6 decreased blood glucose, increased antioxidant enzyme activity, and inhibited glycogen synthesis in T2DM mice. In addition, DhHP-6 improved insulin resistance by activating phosphatidylinositol 3-kinase (PI3K)/AKT, and AMP-activated protein kinase (AMPK) pathway in T2DM mice. Furthermore, DhHP-6 also activated PI3K/AKT and AMPK pathway in glucosamine-induced HepG2 cells. However, LY294002 did not completely inhibit AKT phosphorylation, and partially inhibited AMPK phosphorylation, whilst compound C only partially reduced AMPK phosphorylation, and also partially inhibited AKT phosphorylation, suggesting that AKT and AMPK interact to improve insulin resistance. Thus, these data suggest that DhHP-6 attenuates insulin resistance via the PI3K/AKT and AMPK pathway.

Dexamethasone and prednisolone treatment in beef cattle: influence on glycogen deposition and gene expression in the liver

The illegal administration of glucocorticoids in livestock is problematic and identification of pathways in which these hormones are involved is critically important, and new direct or indirect biomarkers should be identified. In this work, glucocorticoid transcriptional effects on some genes involved in the glucose metabolism were studied in the bovine liver. This study was conducted on adult Charolais male cattle treated with long-term low dose dexamethasone or prednisolone. Gene expression analysis was conducted in the liver by qPCR, and the geNorm algorithm was applied to select optimal reference genes. In line with the literature, a significant overexpression of genes involved in the gluconeogenic pathway and glycogen synthesis was detected in the liver of dexamethasone-treated animals, but histological and biochemical examination showed hepatocyte glycogen depletion particularly in dexamethasone-treated animals. It possible to hypothesize that glucocorticoids or adrenal insufficiency due to glucocorticoids withdrawal inhibit the enzymatic activity of glycogen synthase and/or induce glycogen autophagy in bovine liver. In fact, markers of glycophagy as starch-binding domain-containing protein 1 and γ-aminobutyric acid receptor-associated protein-like 1 mRNAs were upregulated in the liver by glucocorticoids treatment. Furthermore, glycogen synthase kinase-3 beta gene was significantly overexpressed in dexamethasone-treated animals, and this protein is also implicated in liver autophagy modulation and glycogen synthesis inhibition. These results showed that glucocorticoids likley have dual roles in hepatic glycogen metabolism of cattle, and investigation of these pathways could help find treatment biomarkers.

Circadian clock helps cyanobacteria manage energy in coastal and high latitude ocean.

The circadian clock coordinates cellular functions over the diel cycle in many organisms. The molecular mechanisms of the cyanobacterial clock are well characterized, but its ecological role remains a mystery. We present an agent-based model of Synechococcus (harboring a self-sustained, bona fide circadian clock) that explicitly represents genes (e.g., kaiABC), transcripts, proteins, and metabolites. The model is calibrated to data from laboratory experiments with wild type and no-clock mutant strains, and it successfully reproduces the main observed patterns of glycogen metabolism. Comparison of wild type and no-clock mutant strains suggests a main benefit of the clock is due to energy management. For example, it inhibits glycogen synthesis early in the day when it is not needed and energy is better used for making the photosynthesis apparatus. To explore the ecological role of the clock, we integrate the model into a dynamic, three-dimensional global circulation model that includes light variability due to seasonal and diel incident radiation and vertical extinction. Model output is compared with field data, including in situ gene transcript levels. We simulate cyanobaceria with and without a circadian clock, which allows us to quantify the fitness benefit of the clock. Interestingly, the benefit is weakest in the low latitude open ocean, where Prochlorococcus (lacking a self-sustained clock) dominates. However, our attempt to experimentally validate this testable prediction failed. Our study provides insights into the role of the clock and an example for how models can be used to integrate across multiple levels of biological organization.

Exercise-induced molecular mechanisms promoting glycogen supercompensation in human skeletal muscle

ObjectiveA single bout of exercise followed by intake of carbohydrates leads to glycogen supercompensation in prior exercised muscle. Our objective was to illuminate molecular mechanisms underlying this phenomenon in skeletal muscle of man. MethodsWe studied the temporal regulation of glycogen supercompensation in human skeletal muscle during a 5 day recovery period following a single bout of exercise. Nine healthy men depleted (day 1), normalized (day 2) and supercompensated (day 5) muscle glycogen in one leg while the contralateral leg served as a resting control. Euglycemic hyperinsulinemic clamps in combination with leg balance technique allowed for investigating insulin-stimulated leg glucose uptake under these 3 experimental conditions. Cellular signaling in muscle biopsies was investigated by global proteomic analyses and immunoblotting. We strengthened the validity of proposed molecular effectors by follow-up studies in muscle of transgenic mice. ResultsSustained activation of glycogen synthase (GS) and AMPK in combination with elevated expression of proteins determining glucose uptake capacity were evident in the prior exercised muscle. We hypothesize that these alterations offset the otherwise tight feedback inhibition of glycogen synthesis and glucose uptake by glycogen. In line with key roles of AMPK and GS seen in the human experiments we observed abrogated ability for glycogen supercompensation in muscle with inducible AMPK deletion and in muscle carrying a G6P-insensitive form of GS in muscle. ConclusionOur study demonstrates that both AMPK and GS are key regulators of glycogen supercompensation following a single bout of glycogen-depleting exercise in skeletal muscle of both man and mouse.

Glycogen at the Crossroad of Stress Resistance, Energy Maintenance, and Pathophysiology of Aging.

Glycogen is synthesized and stored to maintain postprandial blood glucose homeostasis and to ensure an uninterrupted energy supply between meals. Although the regulation of glycogen turnover has been well studied, the effects of glycogen on aging and disease development have been largely unexplored. In Caenorhabditis elegans fed a high sugar diet, glycogen potentiates resistance to oxidants, but paradoxically, shortens lifespan. Depletion of glycogen by oxidants or inhibition of glycogen synthesis extends the lifespan of worms by an AMPK-dependent mechanism. Thus, glycogen is not merely an inert storage molecule, but also an active regulator of energy balance and aging. Its depletion by oxidants may be beneficial in the treatment of hyperglycemia and glycogen-related diseases.

Inhibition of Glycogen Synthase II with RNAi Prevents Liver Injury in Mouse Models of Glycogen Storage Diseases.

Glycogen storage diseases (GSDs) of the liver are devastating disorders presenting with fasting hypoglycemia as well as hepatic glycogen and lipid accumulation, which could lead to long-term liver damage. Diet control is frequently utilized to manage the potentially dangerous hypoglycemia, but there is currently no effective pharmacological treatment for preventing hepatomegaly and concurrent liver metabolic abnormalities, which could lead to fibrosis, cirrhosis, and hepatocellular adenoma or carcinoma. In this study, we demonstrate that inhibition of glycogen synthesis using an RNAi approach to silence hepatic Gys2 expression effectively prevents glycogen synthesis, glycogen accumulation, hepatomegaly, fibrosis, and nodule development in a mouse model of GSD III. Mechanistically, reduction of accumulated abnormally structured glycogen prevents proliferation of hepatocytes and activation of myofibroblasts as well as infiltration of mononuclear cells. Additionally, we show that silencing Gys2 expression reduces hepatic steatosis in a mouse model of GSD type Ia, where we hypothesize that the reduction of glycogen also reduces the production of excess glucose-6-phosphate and its subsequent diversion to lipid synthesis. Our results support therapeutic silencing of GYS2 expression to prevent glycogen and lipid accumulation, which mediate initial signals that subsequently trigger cascades of long-term liver injury in GSDs.

Transcriptional Activation of Glycogen Catabolism and the Oxidative Pentose Phosphate Pathway by NrrA Facilitates Cell Survival Under Nitrogen Starvation in the Cyanobacterium Synechococcus sp. Strain PCC 7002.

Cyanobacteria respond to nitrogen deprivation by changing cellular metabolism. Glycogen is accumulated within cells to assimilate excess carbon and energy during nitrogen starvation, and inhibition of glycogen synthesis results in impaired nitrogen response and decreased ability to survive. In spite of glycogen accumulation, genes related to glycogen catabolism are up-regulated by nitrogen deprivation. In this study, we found that glycogen catabolism was also involved in acclimation to nitrogen deprivation in the cyanobacterium Synechococcus sp. PCC 7002. The glgP2 gene, encoding glycogen phosphorylase, was induced by nitrogen deprivation, and its expression was regulated by the nitrogen-regulated response regulator A (NrrA), which is a highly conserved transcriptional regulator in cyanobacteria. Activation of glycogen phosphorylase under nitrogen-deprived conditions was abolished by disruption of the nrrA gene, and survival of the nrrA mutant declined. In addition, a glgP2 mutant was highly susceptible to nitrogen starvation. NrrA also regulated expression of the tal-zwf-opcA operon, encoding enzymes of the oxidative pentose phosphate (OPP) pathway, and inactivation of glucose-6-phosphate dehydrogenase, the first enzyme of the OPP pathway, decreased the ability to survive under nitrogen starvation. It was concluded that NrrA facilitates cell survival by activating glycogen degradation and the OPP pathway under nitrogen-deprived conditions.

Suppression of leptin signaling reduces polyglucosan inclusions and seizure susceptibility in a mouse model for Lafora disease.

Lafora disease (LD) represents a fatal form of neurodegenerative disorder characterized by the presence of abnormally large number of polyglucosan bodies-called the Lafora bodies-in neurons and other tissues of the affected patients. The disease is caused by defects in the EPM2A gene coding for a protein phosphatase (laforin) or the NHLRC1 gene coding for an ubiquitin ligase (malin). Studies have shown that inhibition of glycogen synthesis in the brain could prevent the formation of Lafora bodies in the neurons and reduce seizure susceptibility in laforin-deficient mouse, an established animal model for LD. Since increased glucose uptake is thought to underlie increased glycogen in LD, and since the adipocyte hormone leptin is known to positively regulate the glucose uptake in neurons, we reasoned that blocking leptin signaling might reduce the neuronal glucose uptake and ameliorate the LD pathology. We demonstrate here that mice that were deficient for both laforin and leptin receptor showed a reduction in the glycogen level, Lafora bodies and gliosis in the brain, and displayed reduced susceptibility to induced seizures as compared to animals that were deficient only for laforin. Thus, blocking leptin signaling could be a one of the effective therapeutic strategies in LD.