Rabbit brain glucose transporter responds to insulin when expressed in insulin-sensitive Chinese hamster ovary cells.
Transfection of Chinese hamster ovary cells with the expression vector containing rabbit brain HepG2-type glucose transporter cDNA resulted in a dramatic over-expression (approximately 10-fold) of glucose transporter as assessed by either immunoblotting with antipeptide antibody against rabbit brain glucose transporter or photoaffinity labeling with [3H]cytochalasin B. 2-Deoxyglucose uptake was also increased 4-fold in the transfected cells, while no increase in transport activity or transporter amount was observed in cells that were transfected with the expression vector alone without glucose transporter cDNA. Significantly, insulin (10(-7) M) increased 2-deoxyglucose uptake in both control and transfected cells, but the increased amount of the transported 2-deoxyglucose by insulin in the transfected cells was 4.2-fold greater than that in control cells, indicating that the expressed rabbit brain HepG2-type glucose transporter responded to insulin. In addition, we have recently demonstrated that the HepG2-type glucose transporter exists in rat adipocytes and responds to insulin in a fashion similar to a majority of other types of glucose transporters (Oka, Y., Asano, T., Shibasaki, Y., Kasuga, M., Kanazawa, Y., and Takaku, F. (1988) J. Biol. Chem. 263, 13432-13439). In contrast, insulin did not stimulate glucose transport activity in HepG2 cells or IM-9 lymphocytes that have a significant amount of the HepG2-type glucose transporter. Thus, the results in this study further support the notion that insulin regulation of glucose transport activity depends on a tissue-specific signaling mechanism.
Asano
T
T
Institute for Diabetes Care and Research, Asahi Life Foundation, Tokyo, Japan.
Shibasaki
Y
Y
Ohno
S
S
Taira
H
H
Lin
J L
JL
Kasuga
M
M
Kanazawa
Y
Y
Akanuma
Y
Y
Takaku
F
F
Oka
Y
Y
D000345:Affinity Labels; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D015153:Blotting, Western; D001923:Brain Chemistry; D002460:Cell Line; D006224:Cricetinae; D003571:Cytochalasin B / Q000378:metabolism; D004247:DNA / Q000235:genetics; D003847:Deoxyglucose / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005786:Gene Expression Regulation; D007328:Insulin / Q000494:pharmacology*; D008970:Molecular Weight; D009004:Monosaccharide Transport Proteins / Q000235:genetics / Q000378:metabolism*; D010053:Ovary; D010777:Photochemistry; D011817:Rabbits; D014162:Transfection
D000345:Affinity Labels; D007328:Insulin; D009004:Monosaccharide Transport Proteins; D003571:Cytochalasin B; D004247:DNA; D003847:Deoxyglucose