• The Korean Journal of Physiology
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• 제24권2호
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• pp.331-344
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• 1990

What makes glucose transport function sensitive to insulin in one cell type such as adipocyte, and insensitive in another such as liver cells is unresolved question at this time. Recently it is known that insulin stimulates glucose transport in adipocytes largely by redistributing transporter from the storage pool that is included in a low density microsomal fraction to plasma membrane. Therefore, insulin sensitivity may depend upon the relative distribution of gluscose transporters between the plasma membrane and in an intracellular storage compartment. In hepatocytes, the subcellular distribution of glucose transporter is less well documented. It is thus possible that the apparent insensitivity of the hepatocyte system could be either due to lack of the constitutively maintained, intracellular storage pool of glucose transporter or lack of insulin-mediated transporter translocation mechanism in this cell. In this study, I examined if any intracellular glucose transporter pool exists in hepatocytes and this pool is affected by insulin. The results obtained summarized as followings: 1) Distribution of subcellular fractions of hepatocyte showed that there are $24.9{\pm}1.3%$ of plasma membrane, $36.9{\pm}1.7%$ of nucleus-mitochondria enriched fraction, $23.5{\pm}1.2%$ of lysosomal fraction, $9.6{\pm}1.0%$ of high density microsomal fraction and $4.9{\pm}0.5%$ of low density microsomal fraction. 2) In adipocyte, there were $29.9{\pm}2.6%$ of plasma membrane, $19.4{\pm}1.9%$ of nucleus-mitochondria enriched fraction, $26.7{\pm}1.8%$ of high density microsomal fraction and $23.9{\pm}2.1%$ of low density microsomal fraction. 3) Surface labelling of sodium borohydride revealed that plasma membrane contaminated to lysosomal fraction by $26.8{\pm}2.8%$, high density microsomal fraction by $8.3{\pm}1.3%$ and low density microsomal fraction by $1.7{\pm}0.4%$ respectively. 4) Cytochalasin B bound to all of subcellular fractions with a Kd of $1.0{\times}10^{-6}M$. 5) Photolabelling of cytochalasin B to subcellular fractions occurred on 45 K dalton protein band, a putative glucose transporter and D-glucose inhibited the photolabelling. 6) Insulin didn't affect on the distribution of subcellular fractions and translocation of intracellular glucose transporters of hepatocytes. 7) HEGT reconstituted into hepatocytes was largely associated with plasma membrane and very little was found in low density microsomal fraction which equals to the native glucose transporter distribution. Insulin didn't affect on the distribution of exogeneous glucose transporter in hepatocytes. From the above results it is concluded that insulin insensitivity of hepatocyte may due to lack of intracellular storage pool of glucose transporter and thus intracellular storage pool of glucose transporter is an essential feature of the insulin action.

• 미생물학회지
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• 제16권3호
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• pp.122-130
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• 1978

Saccharomyces cerevisiae J170, matant를 사용하여 YEP 배지에서 20시간 진탕 배양하여 생장이 왕성한 영양시기의 효모세포를 얻었다. 이 세포를 sporulation medium에 접종하여 $30^{\circ}C$에서 20시간 진탕 배양하면 약 80%의 ascospore를 형성함을 볼 수 있었다. 포자행성시기와 생장시기를 택하여 D-$^{14}C$-glucose 운반에 관하여 바교 검도한 결과는 다음파 같다. haploid인 포자형성시나 diploid인 생장시기의 포도당 흡수는 pH6.0에서 최대이었다. 이 두 시기의 포도당 운반은 2, 4-dinitrophenol에 의하여 저해되었으므로 포자형성기의 포도당운반도 역시 egergy를 요구하는 능동수송파 연관이 있음을 알게 되었다. 생장시기의 포도당 흡수정도는 포자형성기 보다 높았으며 이때에 fructose나 galactose와 포도당흡수의 관계는 서로 경쟁적으로 작용함을 알 수 있었다. 효모세포의 당 성분은 ribose, mannose, $\alpha$, $\beta$-glucose로 되어 있으며 특히 포자형성기의 mannose 함량은 영양세포 때보다 현저히 적었다.

• Reproductive and Developmental Biology
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• 제39권4호
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• pp.97-104
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• 2015

Glucose is universal and essential fuel of energy metabolism and in the synthesis pathways of all mammalian cells. Glucose is the one of the major precursors of lactose synthesis using glycolysis result in producing milk fat and protein. During the milk fat synthesis, lipoprotein lipase (LPL) and CD36 are required for glucose uptake. Various morecules such as acyl-CoA synthetase 1 (ACSL1) activity of acetyl-CoA synthetase 2 (ACSS2), ACACA, FASN AGPAT6, GPAM, LPIN1 are closely related with milk fat synthesis. Additionally, glucose plays a major role for synthesizing lactose. Activations of lactose synthesize enzymes such as membranebound enzyme, beta-1,4-galactosyl transferase (B4GALT), glucose-6-phosphate dehydrogenase (G6PD) are changed by concentration of glucose in blood resulting change of amount of lactose production. Glucose transporters are a wide group of membrane proteins that facilitate the transport of glucose over a plasma membrane. There are 2 types of glucose transporters which consisted facilitative glucose transporters (GLUT); and sodium-dependent transport, mediated by the Na+/glucose cotransporters (SGLT). Among them, GLUT1, GLUT8, GLUT12, SGLT1, SGLT2 are main glucose transporters which involved in mammary gland development and milk synthesis. However, more studies are required for revealing clear mechanism and function of other unknown genes and transporters. Therefore, understanding of the mechanisms of glucose usage and its regulation in mammary gland is very essential for enhancing the glucose utilization in the mammary gland and improving dairy productivity and efficiency.

• The Korean Journal of Physiology and Pharmacology
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• 제3권3호
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• pp.357-364
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• 1999

We investigated the role of $Ca^{2+}$ and protein kinases/phosphatases in the stimulatory effect of insulin on glucose transport. In isolated rat adipocytes, the simple omission of $CaCl_2$ from the incubation medium significantly reduced, but did not abolish, insulin-stimulated 2-deoxy glucose (2-DG) uptake. Pre-loading adipocytes with intracellular $Ca^{2+}$ chelator, 5,5'-dimethyl bis (o-aminophenoxy)ethane-N,N,N'N' tetraacetic acetoxymethyl ester (5,5'-dimethyl BAPTA/AM) completely blocked the stimulation. Insulin raised intracellular $Ca^{2+}$ concentration $([Ca^{2+}]_i)$ about 1.7 times the basal level of $72{\pm}5$ nM, and 5,5'-dimethyl BAPTA/AM kept it constant at the basal level. This correlation between insulin-induced increases in 2-DG uptake and $[Ca^{2+}]_i$ indicates that the elevation of $[Ca^{2+}]_i$ may be prerequisite for the stimulation of glucose transport. Studies with inhibitors (ML-9, KN-62, cyclosporin A) of $Ca^{2+}-calmodulin$ dependent protein kinases/phosphatases also indicate an involvement of intracellular $Ca^{2+}.$ Additional studies with okadaic acid and calyculin A, protein phosphatase-1 (PP-1) and 2A (PP-2A) inhibitors, indicate an involvement of PP-1 in insulin action on 2-DG uptake. These results indicate an involvement of $Ca^{2+}-dependent$ signaling pathway in insulin action on glucose transport.

• 한국양식학회:학술대회논문집
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• 한국양식학회 2003년도 추계학술발표대회 논문요약집
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• pp.105-106
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• 2003

Physiological responses (cortisol, glucose, lactic acid, osmolality and hematology) of olive flounder (Paralichthys olivaceus) to stressors associated with confinement and subsequent transport were investigated. Specimens were subjected to confinement stress for 3 h, prior to transport for 15 h. Two different size cohorts of the fish, large (839.6$\pm$162.7 g) and small (98.2$\pm$14.8 g), were used. Experimental cohorts of the fish were divided into 3 groups for blood sampling: group A, sampled at the beginning of confinement and 3 h before transport (BT, -3 h), after confinement and at the beginning of transport (BT, 0 h), 3 h after transport had begun (AT, 3 h), and after 15 h transport (AT, 15 h); group B, sampled at BT, 0 h, at AT, 3 h, and at AT, 15 h; and, group C, sampled at AT, 3 h, and at AT, 15 h. In the cohort of large fish, plasma cortisol levels of the A group were increased over time, from 4.2 ng/ml (BT, -3 h), to 92.0 ng/ml (BT, 0 h), 118.5 ng/ml (AT, 3 h) and 105.5 ng/ml (AT, 15 h). A similar pattern was evident in the B group, in which cortisol increased from 47.5 ng/ml (BT, 0 h) to 53.5 ng/ml (AT, 15 h); and, for the C group, in which cortisol increased from 43.5 ng/ml (AT, 3 h) to 71.5 ng/ml (AT, 15 h). Glucose levels of the A group also were significantly increased, from 39.5 mg/dl (BT, -3 h), to 121.0 mg/dl (BT, 0 h), 298.0 mg/dl (AT, 3 h) and 260.5 mg/dl (AT, 15 h). Lactic acid levels increased markedly during transport, from less than 1 mmol/L (BT, 0 h) to 12.0 mmol/L (AT, 15 h). Plasma osmolality increased from 405.5 mOsm/kg (BT, -3 h, for group A) to values near 500 mOsm/kg subsequent to confinement and transport. In the small-size cohort, plasma cortisol, glucose, lactic acid and osmolality levels showed similar but less pronounced trends than those observed for the large-size cohort. This research provides baseline data on cortisol, glucose, lactic acid, osmolality and hematological responses to confinement and transport, which should be useful to aquaculturists working with olive flounder and to scientists studying other flatfish species.

• Biomolecules & Therapeutics
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• 제24권1호
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• pp.94-98
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• 2016

The objective of the present study was to elucidate the effect of bisphosphonates, anti-osteoporosis agents, on glucose uptake in retinal capillary endothelial cells under normal and high glucose conditions. The change of glucose uptake by pre-treatment of bisphosphonates at the inner blood-retinal barrier (iBRB) was determined by measuring cellular uptake of $[^3H]3$-O-methyl glucose (3-OMG) using a conditionally immortalized rat retinal capillary endothelial cell line (TR-iBRB cells) under normal and high glucose conditions. $[^3H]3$-OMG uptake was inhibited by simultaneous treatment of unlabeled D-glucose and 3-OMG as well as glucose transport inhibitor, cytochalasin B. On the other hand, simultaneous treatment of alendronate or pamidronate had no significant inhibitory effect on $[^3H]3$-OMG uptake by TR-iBRB cells. Under high glucose condition of TR-iBRB cells, $[^3H]3$-OMG uptake was increased at 48 h. However, $[^3H]3$-OMG uptake was decreased significantly by pre-treatment of alendronate or pamidronate compared with the values for normal and high glucose conditions. Moreover, geranylgeraniol (GGOH), a mevalonate pathway intermediate, increased the uptake of $[^3H]3$-OMG reduced by bisphosphonates pre-treatment. But, pre-treatment of histamine did not show significant inhibition of $[^3H]3$-OMG uptake. The glucose uptake may be down regulated by inhibiting the mevalonate pathway with pre-treatment of bisphosphonates in TR-iBRB cells at high glucose condition.

• The Korean Journal of Physiology and Pharmacology
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• 제3권2호
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• pp.191-198
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• 1999

This study was undertaken to examine the effect of ethanol on $Na^+-dependent$ transport systems (glucose, phosphate, and dicarboxylate) in renal brush-border membrane vesicles (BBMV). Ethanol inhibited $Na^+-dependent$ uptakes of glucose, phosphate, and succinate in a dose-dependent manner, but not the uptakes of $Na^+-dependent.$ The $H^+/TEA$ antiport was reduced by 8% ethanol. Kinetic analysis showed that ethanol caused a decrease in $V_{max}$ of three transport systems, leaving $K_m$ values unchanged. Ethanol decreased phlorizin binding, which was closely correlated with the decrease in $V_{max}$ of $Na^+-glucose$ uptake. These results indicate that ethanol inhibits $Na^+-dependent$ uptakes of glucose, phosphate, and dicaboxylate and that the reduction in $V_{max}$ of $Na^+-glucose$ uptake is caused by a decrease in the number of active carrier proteins in the membrane.

• 한국양식학회지
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• 제16권3호
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• pp.135-141
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• 2003

활어수송전의 가두기와 활어수송 스트레스에 대한 넙치의 생리학적 반응을 조사하고자, 수송전후의 채혈시각에 따라 3그룹으로 나누어 실험하였다. 실험어류로 양식장에서 사육중인 넙치 성어(839.6$\pm$ 162.7 g, 넙치대)와 유어(98.2$\pm$14.8 g, 넙치소)를 이용하였다. 수송은 제주에서 부산까지 15시간 실시하였다. 넙치대의 cortisol농도는 수송 3시간전 4.2 ng/ml로부터 A그룹은 3시간 가두기후 92.0 ng/ml, 수송중 118.5 ng/ml, 수송직후 105.5 ng/ml로 차이를 나타냈다. B그룹은 3시간 가두기후 47.5 ng/ml, 수송직후 53.5 ng/ml로 높아졌다. C그룹은 수송중 43.5 ng/ml로부터 수송직후 71.5 ng/ml로 더욱 높아졌다. 넙치대치 glucose 농도는 수송 3시간전 39.5 mg/dl로부터 A그룹은 3시간 가두기후 121.0 mg/dl, 수송중 298.0 mg/dl, 수송직후 260.5 mg/dl로 높아졌다. Lactic acid농도는 넙치대에서 수송 3시간전 0.6 mmol/L로부터 수송직후에 A, B 및 C의 모든 그룹에서 0.9~12.0 mmol/L로 높아진 반면, 넙치소에서는 수송 3시간전 1.0 mmol/L로부터 수송직후까지 0.6~1.5 mmol/L로 낮았다. 넙치대의 삼투질 농도는 수송 3시간전 405.5 mOsm/kg으로부터 A그룹은 496.0~526.0 mOsm/kg, B그룹은 479.0~524.5 mOsm/kg, C그룹은 520.5~529.0 mOsm/kg으로 높아졌다. 결론적으로, 가두기 및 수송각업은 넙치의 혈장 cortisol, glucose, 삼투질 농도, ALT, hematocrit, 적혈구수 및 hemoglobin에 영향을 미쳤으며, 넙치소의 스트레스 반응은 넙치대 보다 적게 나타났다. 이상의 결과는 가두기와 활어수송시 혈액내 cortisol, glucose, lactic acid, 삼투질 농도 및 혈액의 일반성상에 대한 기준치를 제시함으로써, 넙치의 생산성 향상과 가자미류의 생리연구에 활용될 것이다.

• Journal of Animal Science and Technology
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• 제51권6호
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• pp.493-502
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• 2009

Glucose is a major source of metabolic fuel and lipid and protein syntheses. Transport of glucose into the cell is regulated by an action of glucose transport.associated transporters, especially solute carriers 2A (Slc2a, protein symbol GLUT). The present study was focused on examination of mRNA expression of various Slc2a isoforms in the epididymis during postnatal development. Total RNAs isolated from different epididymal segments (caput, corpus, and caudal epididymis) were utilized for real-time polymerase chain reaction analyses. Results showed that Slc2a 1, 3, 4, 5, and 8 were expressed in the entire epididymal regions. In addition, the abundance of these Slc2a isoforms' transcripts was different within each epididymal regions. Moreover, the present study showed differential expression of these Slc2a isoforms among different epididymal segments according to postnatal ages. The current study suggests that glucose transport in the epididymis via various Slc2a isoforms would be necessary for maintenance of the epididymal functions.

• Preventive Nutrition and Food Science
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• 제7권2호
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• pp.113-118
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• 2002

Alpha-lipoic acid is a known hypoglycemic agent that may be useful in the treatment of diabetes. The objective of this study was to investigate the fate of glucose in isolated muscles incubated with lipoic acid by determining its direct effects on specific metabolic and signaling pathways. Soleus muscles from healthy rats were incubated with lipoic acid in the absence or presence of insulin. Glucose transport, glycogen synthesis, glucose oxidation and lipid synthesis were determined and affects on major pathways associated with insulin signaling were evaluated. Glucose transport was not significantly altered by the addition of lipoic acid to the incubation medium. However, lipoic acid decreased glycogen synthesis in comparison to controls. Glucose oxidation was moderately increased while de-novo lipid synthesis from glucose was inhibited. Wortmannin repressed insulin stimulation of glucose incorporation into glycogen, an effect that was augmented by the combined treatment of wortmannin and lipoic acid. Basal and insulin-stimulated serine phosphorylation of Akt was not changed by the addition of lipoic acid to the incubation medium. These data show that in this in vitro model, lipoic acid did not significantly affect glucose uptake but dramatically modified pathways of glucose metabolism within muscle tissue.