Differential Expression Research Of Glucose Transporter 4 And Glucose Transporter 1 In Rana Dybowskii During Cold Exposure

Northeast forest frog(Rana dybowskii) in China are mainly distributed in the northeast area, the area in the sub boreal, where winter persistent more than three months. the northeast forest frog caused severe threats to survival. In response to the cold exposure, the northeast forest frog will accumulate high concentration of glucose in the body as cryoprotectance. The glucose transporter is responsible for glucose transport across the cell membrane, one of GLUTs, GLUT4 mainly exists in adipose tissue and muscle tissue, GLUT1 is currently known distribution in the widest range of glucose transporter, GLUT4 and GLUT1 are both able to specific identification of glucose, and intracellular transport.In this experiment, the R. dybowskii of the dominant species in Northeast China was used as the test material, and the-1℃ was used as the control group, the experiment group at 4℃. In the cold stress 4h,8h,12h,24h,48h and 72h respectively take samples from the liver, heart, skin, skeletal muscle, fat body as the test sample, and analysis of changes in the expression of GLUT4 and GLUT1 mRNA levels, to explore the R. dybowskii how to adapt to the dynamic changes of the glucose transporter in organizations during cold exposure.The main steps of this experiment:1. Cold stress for R. dybowskii.2. Sampling of tissues at different time points during cold stress.3.RACE method was used to obtain the full length sequence of cDNA GLUT4, and bioinformatics analysis was carried out to further understand the structural characteristics.4. Testing the distribution of GLUT4 in the R. dybowskii.5. The expression of GLUT1 and mRNA GLUT4 in different tissues with different time points was analyzed by real-time fluorescence quantitative PCR technology.R. dybowskii GLUT4 cDNA sequence analysis showed that the gene sequences encoding protein composed of 510 amino acid residues, molecular weight for 55.74 kDa, isoelectric point of 7.65, homology with Xenopus laevis consistency highest 85%. R. dybowskii GLUT4 and GLUT1 mRNA levels expression analysis during cold stress showed that 4h-8h GLUT4 mRNA levels in skin and skeletal muscle have significant change respectively as 3.82 times and 3.07 times of control group; At 48h GLUT4 and GLUT1 mRNA levels had significant changes, including GLUT4 in the heart and adipose body respectively 2.4 times and 2.9 times than that of the control group, GLUT1 in the liver, heart, and fat body were control group of 9.17,5.0,7.2. times, at 72h, in different tissues of GLUT4 and GLUT1 mRNA levels recovered to the initial normal levels. Results suggest that GLUT4 and GLUT1 were significant change during cold exposure of R. dybowskii for glucose transport and accumulation; at 4h-8h GLUT4 mainly participate in the peripheral skin and skeletal muscle glucose transport and accumulation, then at 48h, GLUT4 and GLUT1 participation in liver, heart, and fat body glucose translocation and accumulation, at 72h the glucose in the R. dybowskii complete the glucose re-distribution in organs.This experiment through to cold stress in R. dybowskii GLUT4 and GLUT1 mRNA levels analysis, showed that GLUT4 and GLUT1 play an important role in glucose transporter, promoting glucose re-distribution, and help the body to form cryoprotectance during cold stress.