Effect of Epidermal Growth Factor on Sodium–Dependent &lt;i&gt;L&lt;/i&gt;–Alanine Transport in LLC–PK1 Cells

Nishida, Masashi; Kawakatsu, Hidekazu; Matsumura, Takafumi; Ishiwari, Kohhei; Tamai, Megumi; Sawada, Tadashi

doi:10.1159/000025948

Cited by 8 publications

(6 citation statements)

References 19 publications

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“…All uptake measurements were made in triplicate. Fructose, L-alanine, or L-arginine uptake experiments were conducted as described by the method of Corpe et al [31], Nishida et al [32], or Acevedo et al [33], respectively.…”

Section: Uptake Experiments [mentioning

confidence: 99%

PKC and MAPKs Pathways Mediate EGF-induced Stimulation of 2-Deoxyglucose Uptake in Mouse Embryonic Stem Cells

Heo

Han

2006

Cell Physiol Biochem

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It has been reported that epidermal growth factor (EGF) and EGF receptor were highly expressed in embryo, suggesting that the EGF system is related to early embryo development in an autocrine and/or paracrine manner. Glucose becomes the preimplantation exogenous energy substrate and enters the blastocyst via glucose transporters. Thus, the effect of EGF on [3H]-2-deoxyglucose (2-DG) uptake and its related signaling pathways were examined in mouse embryonic stem (ES) cells. EGF significantly increased 2-DG uptake in time- and concentration- dependent manner (>12 hr, >10 ng/ ml) and increased mRNA and protein level of glucose transporter 1 (GLUT1) compared to control, respectively. Actinomycin D and cycloheximide completely blocked the effect of EGF on 2-DG uptake. EGF-induced increase of 2-DG uptake was blocked by AG1478 (EGF receptor tyrosine kinase blocker), genistein or herbimycin (tyrosine kinase inhibitors). In addition, EGF effect was blocked by neomycin and U 73122 [phospholipase C (PLC) inhibitors] as well as staurosporine and bisindolylmaleimide I [protein kinase C (PKC) inhibitors]. EGF was also observed to increase inositol phosphates (IPs) formation and activate a PKC translocation from the cytosolic to membrane fraction, suggesting a role of PLC and PKC. SB 203580 [p38 mitogen activated protein kinase (MAPK) inhibitor] or PD 98059 (p44/42 MAPKs inhibitor) blocked EGF-induced increase of 2-DG uptake. EGF also increased phosphorylation of p38 MAPK and p44/42 MAPKs, which was blocked by genistein or bisindolylmaleimide I, respectively. In conclusion, EGF partially increased 2-DG uptake via PKC, p38 MAPK, and p44/42 MAPKs in mouse ES cells.

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Section: Uptake Experiments [mentioning

confidence: 99%

PKC and MAPKs Pathways Mediate EGF-induced Stimulation of 2-Deoxyglucose Uptake in Mouse Embryonic Stem Cells

Heo

Han

2006

Cell Physiol Biochem

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“…All uptake measurements were made in triplicate. 3-O-MG, fructose, l-alanine, l-arginine, and Ca 2+ uptake experiments were conducted as described by the method of Bading et al (1996), Corpe et al (1996), Nishida et al (2000), Acevedo et al (2001), and Chirayath et al (1998), respectively.…”

Section: -Dg Uptake Experimentsmentioning

confidence: 99%

ANG II increases 2-deoxyglucose uptake in mouse embryonic stem cells

Han¹,

Heo²,

Lee³

2005

Life Sciences

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“…All uptake measurements were made in triplicate. 3‐ O ‐Methylglucose, fructose, l ‐alanine, l ‐arginine and Ca 2+ uptake experiments were conducted as described by Bading et al ., 20 Corpe et al ., 21 Acevedo et al ., 22 Nishida et al 23 . and Rabito et al ., 24 respectively.…”

Section: Methodsmentioning

confidence: 99%

HIGH GLUCOSE‐INDUCED INHIBITION OF 2‐DEOXYGLUCOSE UPTAKE IS MEDIATED BY cAMP, PROTEIN KINASE C, OXIDATIVE STRESS AND MITOGEN‐ACTIVATED PROTEIN KINASES IN MOUSE EMBRYONIC STEM CELLS

Han¹,

Heo²,

Lee³

et al. 2006

Clin Exp Pharma Physio

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Abnormally high glucose levels may play an important role in early embryo development and function. In the present study, we investigated the effect of high glucose on 2-deoxyglucose (2-DG) uptake and its related signalling pathway in mouse embryonic stem (ES) cells. 2. 2-Deoxyglucose uptake was maximally inhibited by 25 mmol/L glucose after 24 h treatment. However, 25 mmol/L mannitol and dextran did not affect 2-DG uptake. Indeed, 25 mmol/L glucose decreased GLUT-1 mRNA and protein levels. The glucose (25 mmol/L)-induced inhibition of 2-DG uptake was blocked by pertussis toxin (a G(i)-protein inhibitor; 2 ng/mL), SQ 22,536 (an adenylate cyclase inhibitor; 10(-6) mol/L) and the protein kinase (PK) A inhibitor myristoylated PKI amide-(14-22) (10(-6) mol/L). Indeed, 25 mmol/L glucose increased intracellular cAMP content. 3. Furthermore, 25 mmol/L glucose-induced inhibition of 2-DG uptake was prevented by 10(-4) mol/L neomycin or 10(-6) mol/L U 73,122 (phospholipase C (PLC) inhibitors) and staurosporine or bisindolylmaleimide I (protein kinase (PK) C inhibitors). At 25 mmol/L, glucose increased translocation of PKC from the cytoplasmic fraction to the membrane fraction. The 25 mmol/L glucose-induced inhibition of 2-DG uptake and GLUT-1 protein levels was blocked by SQ 22,536, bisindolylmaleimide I or combined treatment. In addition, 25 mmol/L glucose increased cellular reactive oxygen species and the glucose-induced inhibition of 2-DG uptake were blocked by the anti-oxidants N-acetylcysteine (NAC; 10(-5) mol/L) or taurine (2 yen 10(-3) mol/L). 4. Glucose (25 mmol/L) activated p38 mitogen-activated protein kinase (MAPK) and p44/42 MAPK. Staurosporine (10(-6) mol/L), NAC (10(-5) mol/L) and PD 98059 (10(-7) mol/L) attenuated the phosphorylation of p44/42 MAPK. Both SB 203580 (a p38 MAPK inhibitor; 10(-7) mol/L) and PD 98059 (a p44/42 MAPK inhibitor; 10(-7) mol/L) blocked 25 mmol/L glucose-induced inhibition of 2-DG uptake. 5. In conclusion, high glucose inhibits 2-DG uptake through cAMP, PLC/PKC, oxidative stress or MAPK in mouse ES cells.

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Effect of Epidermal Growth Factor on Sodium–Dependent <i>L</i>–Alanine Transport in LLC–PK1 Cells

Cited by 8 publications

References 19 publications

PKC and MAPKs Pathways Mediate EGF-induced Stimulation of 2-Deoxyglucose Uptake in Mouse Embryonic Stem Cells

PKC and MAPKs Pathways Mediate EGF-induced Stimulation of 2-Deoxyglucose Uptake in Mouse Embryonic Stem Cells

ANG II increases 2-deoxyglucose uptake in mouse embryonic stem cells

HIGH GLUCOSE‐INDUCED INHIBITION OF 2‐DEOXYGLUCOSE UPTAKE IS MEDIATED BY cAMP, PROTEIN KINASE C, OXIDATIVE STRESS AND MITOGEN‐ACTIVATED PROTEIN KINASES IN MOUSE EMBRYONIC STEM CELLS

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