Glucose transporters of rat peripheral nerve. Differential expression of GLUT1 gene by Schwann cells and perineural cells in vivo and in vitro.
Expression of GLUTs in rat peripheral nerve was first studied at the mRNA level with Northern transfer analysis with cDNAs specific for GLUT1, GLUT2, GLUT3, and GLUT4. GLUT1 mRNA was the only GLUT mRNA detectable in rat sciatic nerve. In situ hybridization localized this mRNA to the perineurium and to some endo- and epineurial capillaries. Indirect immunofluorescence stainings demonstrated that GLUT1 protein epitopes were concentrated primarily in the perineurium and endoneurial capillaries. Also, some Schwann cells, a few epineurial capillaries, and medium-sized blood vessels showed a faintly positive immunoreaction. All cell types present in primary cultures initiated from rat sciatic nerve (perineurial cells, Schwann cells, and fibroblasts) expressed GLUT1 protein in vitro. Thus, Schwann cells, which expressed GLUT1 only occasionally at a low level in vivo, have the potential to express GLUT1 at a markedly higher level under cell culture conditions. Incubation of the cultures in 25 mM D-glucose for 7 days caused a 39% reduction in the amount of immunodetectable GLUT1 protein, and a marked (34%) decrease of GLUT1 mRNA compared with cultures incubated in 5.5 mM D-glucose. Interestingly, the reduction of [3H]-2-DG uptake in the same cultures exceeded 70%, suggesting that the reduced amount of GLUT1 protein alone did not explain the marked reduction in glucose uptake in these cultures. Immunostaining of the cell cultures suggested that perineurial cells were the main target for the glucose-induced decrease of GLUT1 protein.
Muona
P
P
Department of Medical Biochemistry, University of Turku, Finland.
Sollberg
S
S
Peltonen
J
J
Uitto
J
J
D000818:Animals; D001692:Biological Transport; D015152:Blotting, Northern; D015153:Blotting, Western; D002478:Cells, Cultured; D003238:Connective Tissue / Q000378:metabolism*; D015342:DNA Probes; D003922:Diabetes Mellitus, Type 1 / Q000378:metabolism*; D015870:Gene Expression; D005947:Glucose / Q000378:metabolism*; D017403:In Situ Hybridization; D007700:Kinetics; D008297:Male; D008970:Molecular Weight; D009004:Monosaccharide Transport Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D009928:Organ Specificity; D015347:RNA Probes; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D011913:Rats, Inbred BB; D017207:Rats, Sprague-Dawley; D012583:Schwann Cells / Q000378:metabolism*; D012584:Sciatic Nerve / Q000378:metabolism*
D015342:DNA Probes; D009004:Monosaccharide Transport Proteins; D015347:RNA Probes; D012333:RNA, Messenger; D005947:Glucose