Differential glycolytic and glycogenogenic transduction pathways in male and female bovine embryos produced in vitro

2012 ◽  
Vol 24 (2) ◽  
pp. 344 ◽  
Author(s):  
M. Garcia-Herreros ◽  
I. M. Aparicio ◽  
D. Rath ◽  
T. Fair ◽  
P. Lonergan
Keyword(s):  
Glucose Metabolism ◽  
Protein Expression ◽  
Glucose Uptake ◽  
Glucose Transporter ◽  
Glycogen Synthase ◽  
Bovine Embryos ◽  
High Concentration ◽  
Glucose Transporter 1 ◽  
Male And Female ◽  
Glut 1

Previous studies have shown that developmental kinetic rates following IVF are lower in female than in male blastocysts and that this may be related to differences in glucose metabolism. In addition, an inhibition of phosphatidylinositol 3-kinase (PI3-K) inhibits glucose uptake in murine blastocysts. Therefore, the aim of this study was to identify and compare the expression of proteins involved in glucose metabolism (hexokinase-I, HK-I; phosphofructokinase-1, PFK-1; pyruvate kinase1/2, PK1/2; glyceraldehyde-3-phosphate dehydrogenase, GAPDH; glucose transporter-1, GLUT-1; and glycogen synthase kinase-3, GSK-3) in male and female bovine blastocysts to determine whether PI3-K has a role in the regulation of the expression of these proteins. Hexokinase-I, PFK-1, PK1/2, GAPDH and GLUT-1 were present in bovine embryos. Protein expression of these proteins and GSK-3 was significantly higher in male compared with female blastocysts. Inhibition of PI3-K with LY294002 significantly decreased the expression of HK-I, PFK-1, GAPDH, GSK-3 A/B and GLUT-1. Results showed that the expression of glycolytic proteins HK-I, PFK-1, GAPDH and PK1/2, and the transporters GLUT-1 and GSK-3 is regulated by PI3-K in bovine blastocysts. Moreover, the differential protein expression observed between male and female blastocysts might explain the faster developmental kinetics seen in males, as the expression of main proteins involved in glycolysis and glycogenogenesis was significantly higher in male than female bovine embryos and also could explain the sensitivity of male embryos to a high concentration of glucose, as a positive correlation between GLUT-1 expression and glucose uptake in embryos has been demonstrated.

103 TRANSDUCTION PATHWAYS RELATED TO GLUCOSE METABOLISM IN MALE AND FEMALE BOVINE EMBRYOS PRODUCED IN VITRO

2011 ◽  
Vol 23 (1) ◽  
pp. 157
Author(s):  
M. Garcia-Herreros ◽  
I. M. Aparicio ◽  
D. Rath ◽  
T. Fair ◽  
P. Lonergan
Keyword(s):  
Glucose Metabolism ◽  
Protein Expression ◽  
Sodium Dodecyl ◽  
Bovine Embryo ◽  
Bovine Embryos ◽  
Glucose Transporter 1 ◽  
Male And Female ◽  
Glut 1 ◽  
Kinetic Rates

Glucose metabolism plays an important role in energy balance control in mammalian cells and has been widely used as an indicator of embryo developmental competence. Previous studies have shown that developmental kinetic rates following IVF are lower in female than in male blastocysts, which may be related to differences in glucose consumption and metabolism. In addition, we have demonstrated that inhibition of phosphatidylinositol 3-kinase (PI3-K) with a structurally unrelated inhibitor, LY294002, suggests a negative role for PI3-K in the regulation of bovine embryo development (Aparicio et al. 2010 Reproduction 140, 83–92). The aim of this study was to determine whether PI3-K has a role in the regulation of glucose metabolism in Day 7 bovine blastocysts and to study the possible differential protein expression involved in glucose metabolism [hexokinase-I (HK-I), phosphofructokinase-1 (PFK-1), pyruvate kinase1/2 (PMK1/2), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), lactate dehydrogenase A/C (LDHA/C), glucose transporter-1 (GLUT-1) and glycogen synthase kinase-3 (GSK-3A/B)] between in vitro produced male and female embryos derived from IVF with either X- or Y-sorted semen. Day 7 blastocysts derived from unsorted semen (n = 25 blastocysts per group) were incubated up to 12 h in SOF culture medium in the presence or absence of LY294002 (10 μM) and stored. Similarly, male and female Day 7 blastocysts derived from sorted semen were collected apart and stored at –80°C until proteomic analysis (Western blot analysis of proteins separated by sodium dodecyl sulfate-polyacylamide gel electrophoresis). Inhibition of PI3K significantly decreased HK-I (P < 0.01), PFK-1 (P < 0.001), GAPDH (P < 0.05), GSK-3A/B (P < 0.001), and GLUT-1 (P < 0.01) protein levels. Interestingly, protein expression of HK-1 (P < 0.001), PFK-1 (P < 0.01), PMK1/2 (P < 0.05), GAPDH (P < 0.01), and GLUT-1 (P < 0.001) was significantly higher in male compared with female blastocysts. The significant increase in the phosphorylated forms (Ser21 and Ser9) of both isoforms (GSK-3A/B) in male compared with female embryos is indicative of a higher inactivation of GSK-3A/B in males (P < 0.001). The presence of LDHA/C activity was not detected in any blastocyst group, irrespective of the gender or treatment studied. In conclusion, our data suggest that PI3K plays a major role in the regulation of glucose metabolism in bovine embryos, because pretreatment with LY294002 significantly modified the protein expression of HK-I, PFK-1, GAPDH, GSK- 3A/B, and GLUT-1, and underline the possibility of modulating glucose metabolism via the PI3K cellular pathway. The differential glycolytic metabolism between male and female blastocysts might explain the higher developmental kinetic rates in males described by other authors, because the expression of proteins involved in glycolysis and glycogenogenesis was significantly higher in male than in female in vitro-produced bovine embryos.

scholarly journals Ecklonia cavaInhibits Glucose Absorption and Stimulates Insulin Secretion in Streptozotocin-Induced Diabetic Mice

2012 ◽  
Vol 2012 ◽  
pp. 1-7 ◽  
Author(s):  
Hye Kyung Kim
Keyword(s):  
Insulin Secretion ◽  
Protein Expression ◽  
Glucose Uptake ◽  
Glucose Transporter ◽  
Islet Cells ◽  
Glucose Absorption ◽  
Oral Glucose ◽  
Diabetic Mice ◽  
Glucose Transporter 1

Aims of study. Present study investigated the effect ofEcklonia cava(EC) on intestinal glucose uptake and insulin secretion.Materials and methods. Intestinal Na+-dependent glucose uptake (SGU) and Na+-dependent glucose transporter 1 (SGLT1) protein expression was determined using brush border membrane vesicles (BBMVs). Glucose-induced insulin secretion was examined in pancreatic β-islet cells. The antihyperglycemic effects of EC, SGU, and SGLT1 expression were determined in streptozotocin (STZ)-induced diabetic mice.Results. Methanol extract of EC markedly inhibited intestinal SGU of BBMV with the IC50value of 345 μg/mL. SGLT1 protein expression was dose dependently down regulated with EC treatment. Furthermore, insulinotrophic effect of EC extract was observed at high glucose media in isolated pancreatic β-islet cellsin vitro. We next conducted the antihyperglycemic effect of EC in STZ-diabetic mice. EC supplementation markedly suppressed SGU and SGLT1 abundance in BBMV from STZ mice. Furthermore, plasma insulin level was increased by EC treatment in diabetic mice. As a result, EC supplementation improved postprandial glucose regulation, assessed by oral glucose tolerance test, in diabetic mice.Conclusion. These results suggest that EC play a role in controlling dietary glucose absorption at the intestine and insulinotrophic action at the pancreas contributing blood glucose homeostasis in diabetic condition.

scholarly journals Glucose uptake inhibition decreases expressions of receptor activator of nuclear factor-kappa B ligand (RANKL) and osteocalcin in osteocytic MLO-Y4-A2 cells

2018 ◽  
Vol 314 (2) ◽  
pp. E115-E123 ◽  
Author(s):  
Ayumu Takeno ◽  
Ippei Kanazawa ◽  
Masakazu Notsu ◽  
Ken-ichiro Tanaka ◽  
Toshitsugu Sugimoto
Keyword(s):  
Glucose Metabolism ◽  
Glucose Uptake ◽  
P38 Mapk ◽  
Glucose Transporter ◽  
Mek Inhibitor ◽  
Long Bone ◽  
Uptake Inhibition ◽  
Glucose Transporter 1 ◽  
P38 Inhibitor ◽  
Glucose Levels

Bone and glucose metabolism are closely associated with each other. Both osteoblast and osteoclast functions are important for the action of osteocalcin, which plays pivotal roles as an endocrine hormone regulating glucose metabolism. However, it is unknown whether osteocytes are involved in the interaction between bone and glucose metabolism. We used MLO-Y4-A2, a murine long bone-derived osteocytic cell line, to investigate effects of glucose uptake inhibition on expressions of osteocalcin and bone-remodeling modulators in osteocytes. We found that glucose transporter 1 (GLUT1) is expressed in MLO-Y4-A2 cells and that treatment with phloretin, a GLUT inhibitor, significantly inhibited glucose uptake. Real-time PCR and Western blot showed that phloretin significantly and dose-dependently decreased the expressions of RANKL and osteocalcin, whereas osteoprotegerin or sclerostin was not affected. Moreover, phloretin activated AMP-activated protein kinase (AMPK), an intracellular energy sensor. Coincubation of ara-A, an AMPK inhibitor, with phloretin canceled the phloretin-induced decrease in osteocalcin expression, but not RANKL. In contrast, phloretin suppressed phosphorylation of ERK1/2, JNK, and p38 MAPK, and treatments with the p38 inhibitor SB203580 and the MEK inhibitor PD98059, but not the JNK inhibitor SP600125, significantly decreased expressions of RANKL and osteocalcin. These results indicate that glucose uptake by GLUT1 is required for RANKL and osteocalcin expressions in osteocytes, and that inhibition of glucose uptake decreases their expressions through AMPK, ERK1/2, and p38 MAPK pathways. These findings suggest that lowering glucose uptake into osteocytes may contribute to maintain blood glucose levels by decreasing osteocalcin expression and RANKL-induced bone resorption.

scholarly journals GLUT-1 Enhances Glycolysis, Oxidative Stress, and Fibroblast Proliferation in Keloid

Life ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 505
Author(s):  
Ying-Yi Lu ◽  
Chieh-Hsin Wu ◽  
Chien-Hui Hong ◽  
Kee-Lung Chang ◽  
Chih-Hung Lee
Keyword(s):  
Glucose Metabolism ◽  
Glucose Transporter ◽  
Metabolic Reprogramming ◽  
Skin Tumor ◽  
Fibroblast Proliferation ◽  
Rt Pcr ◽  
Extracellular Acidification ◽  
Glucose Transporter 1 ◽  
Glut 1 ◽  
Keloid Fibroblasts

A keloid is a fibroproliferative skin tumor. Proliferating keloid fibroblasts (KFs) demand active metabolic utilization. The contributing roles of glycolysis and glucose metabolism in keloid fibroproliferation remain unclear. This study aims to determine the regulation of glycolysis and glucose metabolism by glucose transporter-1 (GLUT-1), an essential protein to initiate cellular glucose uptake, in keloids and in KFs. Tissues of keloids and healthy skin were explanted for KFs and normal fibroblasts (NFs), respectively. GLUT-1 expression was measured by immunofluorescence, RT-PCR, and immunoblotting. The oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) were measured with or without WZB117, a GLUT-1 inhibitor. Reactive oxygen species (ROS) were assayed by MitoSOX immunostaining. The result showed that glycolysis (ECAR) was enhanced in KFs, whereas OCR was not. GLUT-1 expression was selectively increased in KFs. Consistently, GLUT-1 expression was increased in keloid tissue. Treatment with WZB117 abolished the enhanced ECAR, including glycolysis and glycolytic capacity, in KFs. ROS levels were increased in KFs compared to those in NFs. GLUT-1 inhibition suppressed not only the ROS levels but also the cell proliferation in KFs. In summary, the GLUT-1-dependent glycolysis and ROS production mediated fibroblast proliferation in keloids. GLUT1 might be a potential target for metabolic reprogramming to treat keloids.

scholarly journals Subtype-dependent difference of glucose transporter 1 and hexokinase II expression in craniopharyngioma: an immunohistochemical study

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Naoto Mukada ◽  
Masahiko Tosaka ◽  
Nozomi Matsumura ◽  
Rei Yamaguchi ◽  
Masanori Aihara ◽  
...  
Keyword(s):  
Glucose Metabolism ◽  
Glucose Transporter ◽  
Immunohistochemical Analysis ◽  
Braf V600e ◽  
Beta Catenin ◽  
Hexokinase Ii ◽  
Glucose Transporter 1 ◽  
Positive Group ◽  
Glut 1 ◽  
Positive Rate

AbstractPapillary craniopharyngiomas are characterized by the BRAF V600E mutation. Enhancement of glucose metabolism may be involved in the downstream of the BRAF V600E mutation in many types of tumors. Glucose metabolism was investigated in craniopharyngioma using immunohistochemical analysis. The study included 29 cases of craniopharyngioma (18 adamantinomatous type [ACP], 11 papillary type [PCP]). Immunohistochemical analysis was performed with anti-glucose transporter-1 (GLUT-1), anti-hexokinase-II (HK-II), anti-BRAF V600E, and anti-beta-catenin antibodies. Expressions of GLUT-1 and HK-II were evaluated using a semiquantitative 4-tiered scale as 0, 1+, 2+, 3+, and divided into negative (0 or 1+) or positive (2+ or 3+) group. GLUT-1 expression level was significantly higher in PCPs than ACPs (0, 1+, 2+, 3+ = 2, 12, 4, 0 cases in ACP, respectively, 0, 1+, 2+, 3+ = 0, 2, 5, 4 in PCP, p = 0.001), and most PCPs were classified into positive group (positive rate, 22.2% [4/18] in ACP, 81.8% [9/11] in PCP; p = 0.003). HK-II expression was also conspicuous in PCPs (0, 1+, 2+, 3+ = 7, 9, 2, 0 cases in ACP, 0, 3, 3, 5 in PCP; p = 0.001), and most of them divided into positive group (positive rate, 11.1% [2/18] in ACP, 72.7% [8/11] in PCP; p = 0.001). Expression patterns of BRAF V600E and beta-catenin reflected the clinicopathological subtypes. Both GLUT-1 and HK-II expressions were prominent in PCP. Glucose metabolism might be more enhanced in PCP than ACP. PCP may use the glucose metabolic system downstream of the BRAF V600E mutant protein.

scholarly journals GnRH increases glucose transporter-1 expression and stimulates glucose uptake in the gonadotroph

2011 ◽  
Vol 212 (2) ◽  
pp. 139-147 ◽  
Author(s):  
Valerie M Harris ◽  
Sachin V Bendre ◽  
Francina Gonzalez De Los Santos ◽  
Alemu Fite ◽  
Ahmad El-Yaman El-Dandachli ◽  
...  
Keyword(s):  
Glucose Metabolism ◽  
Mrna Expression ◽  
Glucose Uptake ◽  
Glucose Transporter ◽  
Subcellular Location ◽  
Pcr Analysis ◽  
Glucose Transporter 1 ◽  
Glut1 Protein ◽  
Main Regulator ◽  
Exogenous Treatment

GnRH is the main regulator of the hypothalamic–pituitary–gonadal (H–P–G) axis. GnRH stimulates the pituitary gonadotroph to synthesize and secrete gonadotrophins (LH and FSH), and this effect of GnRH is dependent on the availability of glucose and other nutrients. Little is known about whether GnRH regulates glucose metabolism in the gonadotroph. This study examined the regulation of glucose transporters (Gluts) by GnRH in the LβT2 gonadotroph cell line. Using real-time PCR analysis, the expression ofGlut1, -2, -4, and -8 was detected, butGlut1mRNA expression level was more abundant than the mRNA expression levels ofGlut2, -4, and -8. After the treatment of LβT2 cells with GnRH,Glut1mRNA expression was markedly induced, but there was no GnRH-induction ofGlut2, -4, or -8 mRNA expression in LβT2 cells. The effect of GnRH onGlut1mRNA expression is partly mediated by ERK activation. GnRH increased GLUT1 protein and stimulated GLUT1 translocation to the cell surface of LβT2 cells. Glucose uptake assays were performed in LβT2 cells and showed that GnRH stimulates glucose uptake in the gonadotroph. Finally, exogenous treatment of mice with GnRH increased the expression ofGlut1but not the expression ofGlut2, -4, or -8 in the pituitary. Therefore, regulation of glucose metabolism by GnRH via changes inGlutsexpression and subcellular location in the pituitary gonadotroph reveals a novel response of the gonadotroph to GnRH.

Angiotensin II increases glucose uptake and glucose transporter-1 mRNA levels in astroglia

1995 ◽  
Vol 268 (3) ◽  
pp. E384-E390 ◽  
Author(s):  
W. Tang ◽  
E. M. Richards ◽  
M. K. Raizada ◽  
C. Sumners
Keyword(s):  
Steady State ◽  
Angiotensin Ii ◽  
Glucose Uptake ◽  
Glucose Transporter ◽  
Mrna Levels ◽  
Ang Ii ◽  
At1 Receptors ◽  
Glucose Transporter 1 ◽  
Glut 1 ◽  
Steady State Levels

In this study, we have investigated the effects of angiotensin II (ANG II) on glucose uptake into astroglia cultured from adult rat hypothalamus and brain stem. ANG II (30 min to 4 h; 10(-9) to 10(-6) M) stimulated time- and concentration-dependent increases in the uptake of 2-deoxy-D-[3H]glucose into cultured astroglia. This effect of ANG II (10(-7) M) is via AT1 receptors and protein kinase C (PKC), since it was inhibited by losartan (10(-6) M) and staurosporine (10(-6) M), respectively. Furthermore, this ANG II action was inhibited by both cycloheximide (1 microgram/ml) and actinomycin D (10(-6) M), indicating that synthesis of new glucose transporters is involved. This was confirmed by the finding that ANG II (30 min to 4 h; 10(-9) to 10(-5) M) stimulated time- and concentration-dependent increases in the steady-state levels of glucose transporter-1 (GLUT-1) mRNA in these cultures. In addition, the increase in steady-state levels of GLUT-1 mRNA elicited by ANG II was mediated by AT1 receptors and PKC. These data suggest that ANG II stimulates glucose uptake into cultured astroglia via a pathway that involves AT1 receptors, PKC, and increased steady-state levels of GLUT-1 mRNA.

scholarly journals Aspirin, a potential GLUT1 inhibitor in a vascular endothelial cell line

Open Medicine ◽  
2019 ◽  
Vol 14 (1) ◽  
pp. 552-560 ◽  
Author(s):  
Yabo Hu ◽  
Xiaohan Lou ◽  
Ruirui Wang ◽  
Chanjun Sun ◽  
Xiaomeng Liu ◽  
...  
Keyword(s):  
Glucose Metabolism ◽  
Glucose Uptake ◽  
Glucose Transporter ◽  
Vascular Endothelial Cells ◽  
Vascular Endothelial Cell ◽  
Vascular Endothelial ◽  
Glucose Transporter 1 ◽  
Glut1 Expression ◽  
Inhibition Of Angiogenesis ◽  
Underlying Mechanisms

AbstractRecent epidemiological and preclinical studies have revealed that aspirin possesses antitumor properties; one of the mechanisms results from inhibition of angiogenesis. However, the underlying mechanisms of such action remain to be elucidated, in particular, the effect of aspirin on glucose metabolism of vascular endothelial cells (ECs) has not yet been reported. Herein, we demonstrate that glucose transporter 1 (GLUT1), a main glucose transporter in ECs, can be down-regulated by aspirin. Exposure to 4-mM aspirin significantly decreased GLUT1 at the mRNA and protein level, resulting in impaired glucose uptake capacity in vascular ECs. In addition, we also showed that exposure to 4-mM aspirin led to an inhibition of intracellular ATP and lactate synthesis in vascular ECs, and a down-regulation of the phosphorylation level of NF-κB p65 was observed. Taken together, these findings indicate 4-mM aspirin inhibits glucose uptake and glucose metabolism of vascular ECs through down-regulating GLUT1 expression and suggest that GLUT1 has potential to be a target for aspirin in vascular ECs.

Temporally differential protein expression of glycolytic and glycogenic enzymes during in vitro preimplantation bovine embryo development

2018 ◽  
Vol 30 (9) ◽  
pp. 1245 ◽  
Author(s):  
Manuel García-Herreros ◽  
Constantine A. Simintiras ◽  
Patrick Lonergan
Keyword(s):  
Embryo Development ◽  
Glucose Transporter ◽  
Developmental Stages ◽  
Glycogen Synthase ◽  
Differentially Expressed ◽  
Bovine Embryo ◽  
Cell Stage ◽  
Glucose Transporter 1 ◽  
Glut 1

Proteomic analyses are useful for understanding the metabolic pathways governing embryo development. This study investigated the presence of enzymes involved in glycolysis and glycogenesis in in vitro-produced bovine embryos at five developmental stages leading up to blastocyst formation. The enzymes examined were: (1) glycolytic: hexokinase-I (HK-I), phosphofructokinase-1 (PFK-1), pyruvate kinase mutase 1/2 (PKM-1/2), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and (2) glycogenic: glycogen synthase kinase-3 isoforms α/ β (GSK-3α/β). Glucose transporter-1 (GLUT-1) was also analysed. The developmental stages examined were: (1) 2–4-cell, (2) 5–8-cell, (3) 16-cell, (4) morula and (5) expanded blastocyst. The enzymes HK-I, PFK-1, PKM-1/2, GAPDH and GLUT-1 were differentially expressed throughout all stages (P < 0.05). GSK-3α and β were also differentially expressed from the 2–4-cell to the expanded blastocyst stage (P < 0.05) and GLUT-1 was identified throughout. The general trend was that the abundance of PFK1, GAPDH and PKM-1/2 decreased whereas HK-I, phospho-GSK3α (P-GSK3α) and P-GSK3β levels increased as the embryo advanced. In contrast, GLUT-1 expression peaked at the 16-cell stage. These data combined suggest that in vitro bovine embryo metabolism switches from being glycolytic-centric to glycogenic-centric around the 16-cell stage, the developmental window also characterised by embryonic genome activation.

scholarly journals Effect of Inhibition of GLUT1 Expression and Autophagy Modulation on the Growth and Migration of Laryngeal Carcinoma Stem Cells under Hypoxic and Low-Glucose Conditions

2020 ◽  
Author(s):  
Shui-Hong Zhou ◽  
Xiao-Hong Chen ◽  
Jia Liu ◽  
Jiang-Tao Zhong ◽  
Jun Fan
Keyword(s):  
Stem Cells ◽  
Glucose Uptake ◽  
Laryngeal Carcinoma ◽  
Glucose Transporter ◽  
Cell Counting ◽  
Glucose Transporter 1 ◽  
Glut 1 ◽  
Low Glucose ◽  
And Migration ◽  
Proliferation And Migration

Abstract Background: Enhanced glucose uptake and autophagy are means by which cells adapt to stressful microenvironments. We investigated the roles of glucose transporter-1 (GLUT-1) and autophagy in laryngeal carcinoma stem cells under hypoxic and low-glucose conditions.Methods: CD133+ Tu212 laryngeal carcinoma stem cells were purified by magnetic-activated cell sorting and subjected to hypoxic and/or low-glucose conditions. Proliferation was evaluated using a cell-counting kit and a clone-formation assay, and migration was evaluated through a Transwell assay. Autophagy was assessed via transmission electron microscopy. GLUT-1 and beclin-1 expression were silenced using an shRNA and autophagy was manipulated using rapamycin, 3-MA, or chloroquine. Gene expression levels were evaluated by quantitative reverse transcription-polymerase chain reaction and protein concentrations were assessing via Western blotting.Results: Compared to CD133– stem cells, CD133+ cells showed increased proliferation and migration, and reduced apoptosis, under hypoxic or low-glucose conditions. They also showed increased expression of GLUT-1 and autophagy markers. Finally, GLUT-1 knockdown or autophagy inhibition reduced their proliferation and migration.Conclusions: Enhanced glucose uptake and autophagy maintain the functions of CD133+ laryngeal carcinoma stem cells under hypoxic and low-glucose conditions.

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