Functional expression of mammalian glucose transporters in Xenopus laevis oocytes: evidence for cell-dependent insulin sensitivity.
We report the functional expression of two different mammalian facilitative glucose transporters in Xenopus oocytes. The RNAs encoding the rat brain and liver glucose transporters were transcribed in vitro and microinjected into Xenopus oocytes. Microinjected cells showed a marked increase in 2-deoxy-D-glucose uptake as compared with controls injected with water. 2-Deoxy-D-glucose uptake increased during the 5 days after microinjection of the RNAs, and the microinjected RNAs were stable for at least 3 days. The expression of functional glucose transporters was dependent on the amount of RNA injected. The oocyte-expressed transporters could be immunoprecipitated with anti-brain and anti-liver glucose transporter-specific antibodies. Uninjected oocytes expressed an endogenous transporter that appeared to be stereospecific and inhibitable by cytochalasin B. This transporter was kinetically and immunologically distinguishable from both rat brain and liver glucose transporters. The uniqueness of this transporter was confirmed by Northern (RNA) blot analysis. The endogenous oocyte transporter was responsive to insulin and to insulinlike growth factor I. Most interestingly, both the rat brain and liver glucose transporters, which were not insulin sensitive in the tissues from which they were cloned, responded to insulin in the oocyte similarly to the endogenous oocyte transporter. These data suggest that the insulin responsiveness of a given glucose transporter depends on the type of cell in which the protein is expressed. The expression of hexose transporters in the microinjected oocytes may help to identify tissue-specific molecules involved in hormonal alterations in hexose transport activity.
Vera
J C
JC
Program in Molecular Biology, Memorial Sloan-Kettering Cancer Research Center, New York, New York 10021.
Rosen
O M
OM
D000818:Animals; D001692:Biological Transport / Q000235:genetics; D003571:Cytochalasin B / Q000378:metabolism; D003847:Deoxyglucose / Q000378:metabolism; D007328:Insulin / Q000502:physiology; D007334:Insulin-Like Growth Factor I / Q000502:physiology; D008845:Microinjections; D009004:Monosaccharide Transport Proteins / Q000096:biosynthesis / Q000235:genetics*; D009865:Oocytes / Q000378:metabolism*; D010957:Plasmids; D012313:RNA / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis; D014982:Xenopus laevis
D007328:Insulin; D009004:Monosaccharide Transport Proteins; D011994:Recombinant Proteins; D003571:Cytochalasin B; D012313:RNA; D007334:Insulin-Like Growth Factor I; D003847:Deoxyglucose