• 한국발생생물학회지:발생과생식
• /
• 제2권2호
• /
• pp.205-212
• /
• 1998

$Na^{+}$이온 비의존적으로 작동하는 포도당 수송체 (glucose transporter 1, Glut1)는 생쥐 배아의 세포막을 경계로 포도당을 수송하는 주요통로이다. 성장인자 가운데 insulin-like growth factor-I (IGF-I)은 생쥐배아에서 포도당의 유입을 증가시키는 것으로 알려져있으나 이러한 효과가 IGF-I 의한 Glut1의 전사조절 효과에 기인한 것인지는 알려져 있지 않다. 본 연구는 포도당과 IGF-I 생쥐의 착상전 배아 발생과 Glut1 발현에 미치는 영향을 조사함으로써 이들에 의한 배발생 조절기작을 이해하고자 시행하였다. 2-세포기 배아는 배양액내 pyruvate 존재하에 포도당의 유무와 관계없이 포배로 발생하였다. IGF-I은 2-세포기에서 체외 발생한 중기포배내 할구수를 유의하게 증가시켰다. 2-세포기부터 체외발생한 상실배의 Glut1 전사체의 양에는 배양액내 포도당의 유무에 따른 차이가 없었으며, IGF-I은 포도당과 무관하게 Glut1의 발현을 증가시켰다. 이러한 결과에서 상실기 생쥐배아의 경우 단순히 포도당의 결핍에 의해 Glut1의 발현이 전사수준에서 촉진되지 않으며, Glut1 발현의 증가는 IGF-I에 의한 배발생 촉진효과와 관련이 있는 것으로 사료된다.

• Clinical and Experimental Reproductive Medicine
• /
• 제25권1호
• /
• pp.77-86
• /
• 1998

The uptake of glucose for metabolism and growth is essential to most animal cells and is mediated by glucose-transporter (GLUT) proteins. The aim of this study was to determine which class of glucose transporter molecules was responsible for uptake of glucose in the mouse early embryo and at which stage the corresponding genes were expressed. In addition, co-culture system with vero cell was used to investigate the effect of the system on GLUT expression. Two-cell stage embryos were collected from the superovulated ICR female and divided into 3 groups. As a control, embryos were cultured in 0.4% BSA-T6 medium which includes glucose. For the experimental groups, embryos were cultured in either co-culture system with vero cells or glucose-free T6 medium supplemented with 0.4% BSA and pyruvate as an energy substrate. 2-cell to blastocyst stage embryos in those groups were respectively collected into microtubes (50 embryos/tube). Total RNA was extracted and RT-PCR was performed. The products were analysed after staining ethidium bromide by 2% agarose gel electrophoresis. Blastocysts were collected from each group at l20hr after hCG injection. They were fixed in 2.5% glutaraldehyde, stained with hoechst, and mounted for observation. In control, GLUT1 was expressed from 4-cell to blastocyst. GLUT2 and GLUT3 were expressed in morula and blastocyst. GLUT4 was expressed in all stages. When embryos were cultured in glucose-free medium, no significant difference was shown in the expression of GLUT1, 2 and 3, compared to control. However GLUT4 was not expressed until morular stage. When embryos were co-cultured with vero cell, there was no significant difference in the expression of GLUT1, 2, 3 and 4 compared to control. To determine cell growth of embryos, the average cell number of blastocyst was counted. The cell number of co-culture ($93.8{\pm}3.1$, n=35) is significantly higher than that of control and glucose-free group ($76.6{\pm}3.8$, n=35 and $68.2{\pm}4.3$, n=30). This study shows that the GLUT genes are expressed differently according to embryo stage. GLUTs were detectable throughout mouse preimplantation development in control and co-culture groups. However, GLUT4 was not detected from 2- to 8-cell stage but detected from morula stage in glucose-free medium, suggested that GLUT genes are expressed autocrinally in the embryo regardless of the presence of glucose as an energy substrate. In addition, co-culture system can increase the cell count of blastocyst but not improve the expression of GLUT. In conclusion, expression of GLUT is dependent on embryo stage in preimplantation embryo development.

• 한국환경성돌연변이발암원학회:학술대회논문집
• /
• 한국환경성돌연변이발암원학회 2004년도 춘계학술대회
• /
• pp.63-63
• /
• 2004

• 대한의생명과학회지
• /
• 제14권2호
• /
• pp.119-122
• /
• 2008

The baculovirus/insect cell expression system is of great value in the study of structure-function relationships in mammalian glucose-transport proteins by site-directed mutagenesis and for the large-scale production of these proteins for mechanistic and biochemical studies. In order to exploit this, the effects of substitution at the highly conserved residue glutamine 282 of the human erythrocyte-type glucose transporter have been examined by in vitro site-directed mutagenesis. The modified human transport protein has been expressed in Spodoptera frugiperda 21 cells by using the recombinant baculovirus AcNPV-GTL. To assess the functional integrity of the expressed transporter, measurements of the transport inhibitor cytochalasin B binding were performed, involving the membranes prepared from 4 days post infection with no virus, with wild-type virus or AcNPV-GTL virus. Data obtained showed that there was little or no D-glucose-inhibitable binding in cells infected with the wild type or no virus. Only the recombinant virus infected cells exhibited specific binding, which is inhibitable by D- but not by L-glucose. However, there was a notable reduction in the affinity for the potent inhibitor cytochalasin B when binding measurements of AcNPV-GTL were compared with those of AcNPV-GT, which has no substitution. It is thus suggested that although the modified and unmodified human transporters differed slightly in their affinity for cytochalasin B, the glutamine substitution did not interfere the heterologous expression of the human transporter in the insect cells.

• Journal of Microbiology and Biotechnology
• /
• 제14권3호
• /
• pp.450-455
• /
• 2004

A reducing glucose-carrying polymer, called poly [3-O-(4'-vinylbenzyl)-D-glucose］(PVG), was interacted with HepG2 cells including a type-l glucose transporter (GLUT-1) on the cell membrane. The cooperative interaction between a number of GLUT-1s and a number of reducing 3-O-methyl-D-glucose moieties on the PVG polymer chain was found to be responsible for the increase in the interaction with HepG2 cells. The affinity between the cells and the PVG was studied using RITC-labeled glycopolymers. The specific interaction between the GLUT-1 on HepG2 cells and the PVG polymer carrying reducing glucose moieties was suppressed by the inhibitors, phloretin, phloridzin, and cytochalasin B. Direct observation by confocal laser microscopy with the use of RITC-labeled PVG and pretreatment of HepG2 cells with the inhibitors demonstrated that the cells interacted with the soluble form of the PVG polymer via GLUT-1, while fluorescence labeling of the cell surface was prevented after pretreatment with the inhibitors of GLUT-1.

• 한국영양학회:학술대회논문집
• /
• 한국영양학회 2002년도 춘계학술대회
• /
• pp.58-59
• /
• 2002

• 한국영양학회:학술대회논문집
• /
• 한국영양학회 2002년도 춘계 심포지움초록
• /
• pp.612-612
• /
• 2002

• Journal of Microbiology and Biotechnology
• /
• 제30권12호
• /
• pp.1944-1949
• /
• 2020

Mutant sugar transporter ScGAL2-N376F was overexpressed in Kluyveromyces marxianus for efficient utilization of xylose, which is one of the main components of cellulosic biomass. K. marxianus ScGal2_N376F, the ScGAL2-N376F-overexpressing strain, exhibited 47.04 g/l of xylose consumption and 26.55 g/l of xylitol production, as compared to the parental strain (24.68 g/l and 7.03 g/l, respectively) when xylose was used as the sole carbon source. When a mixture of glucose and xylose was used as the carbon source, xylose consumption and xylitol production rates were improved by 195% and 360%, respectively, by K. marxianus ScGal2_N376F. Moreover, the glucose consumption rate was improved by 27% as compared to that in the parental strain. Overexpression of both wild-type ScGAL2 and mutant ScGAL2-N376F showed 48% and 52% enhanced sugar consumption and ethanol production rates, respectively, when a mixture of glucose and galactose was used as the carbon source, which is the main component of marine biomass. As shown in this study, ScGAL2-N376F overexpression can be applied for the efficient production of biofuels or biochemicals from cellulosic or marine biomass.

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

• 한국식물생명공학회:학술대회논문집
• /
• 한국식물생명공학회 2005년도 추계학술대회 및 한일 식물생명공학 심포지엄
• /
• pp.9-21
• /
• 2005

To study the biochemical and physiological role of the plastidic glucose transporter (pGlcT) in carbohydrate metabolism, we characterized transgenic plants with mutations in the pGlcT gene (GT), gt-1 and gt-2, as well double mutants of GT and the maltose transporter (MEX1) and GT and the triose phosphate/phosphate translocator (TPT), GT and the cytosolic fructose-1,6-bisphosphatase gene (cFBP), and MEX1 and TPT, gt-1/mex2, gt-1/tpt-2, gt-1/cfbp-1, mex1-1/tpt-2, respectively. Compared to the wild type, all mutants except the gt-1/cfbp-1 mutant lines displayed higher starch accumulation and higher levels of maltose. Starch accumulation is due to a decrease in starch turnover, leading to an imbalance between the rates of synthesis and degradation. Sucrose levels of gt alleles were higher than those in wild-type plants during the light period, suggesting possible nightly supplementation via the maltose transport pathway to maintain proper carbohydrate partitioning in the plant leaves. The gt plants displayed less growth retardation than mex1-1 mutant and gt-1/mex2 double mutant displayed accumulativesevere growth retardation as compared to individual gt-1 and mex1-1 mutants, implying that the maltose transporter-mediated pathway is a major route for carbohydrate partitioning at night. The gt-1/tpt-2, mex1-1/tpt-2 and gt-1/cfbp-1 double mutants had retarded growth and low chlorophyll content to differing degrees, indicating that photosynthetic capacity had diminished. Interestingly, the gt-1/tpt-2 line displayed a glucose-insensitive phenotype and higher germination rates than wild type, suggesting its involvement not only in carbon partitioning, but also in the sugar signaling network of the pGlcT and TPT.