The myo-inositol transport system in confluent fetal-bovine aortic end
othelial cells was characterized after 7-10 days in subculture, at whi
ch time the myo-inositol levels and rates of myo-[2-H-3]-inositol upta
ke and incorporation into phospholipid had reached steady state. Kinet
ic analysis indicated that the uptake occurred by both a high-affinity
transport system with an apparent K(t) of 31 muM and V(max) of 45 pmo
l/min per mg of protein, and a non-saturable low-affinity system. Upta
ke was competitively inhibited by phlorrhizin, with a K(i) of 50 muM;
phloretin was a non-competitive inhibitor, with half-maximal inhibitio
n between 0.2 and 0.5 mM. Glucose was a weak competitive inhibitor, wi
th a K(i) of 37 mM; galactose failed to inhibit uptake. A weak depende
nce on Na+ for the initial rate of uptake was observed at 11 muM myo-i
nositol. When fetal-bovine-serum (FBS)-supplemented medium, which cont
ained 225 muM myoinositol, was used, the cells contained about 200 nmo
l of myoinositol/mg of DNA. With adult-bovine-serum (ABS)supplemented
medium, which contained 13 muM myo-inositol, the cells contained about
110 nmol/mg of DNA. Transport of 11 muM myo-[2-H-3]inositol was 18 an
d 125 pmol/min per mg of DNA for cells grown in FBS and ABS respective
ly. Kinetic analysis showed that for the cells grown in FBS the V(max)
of the high-affinity system was decreased by 64%, whereas the K(t) re
mained essentially unchanged. Increased cell myo-inositol levels were
not associated with an increased rate of phosphatidylinositol synthesi
s. After prolonged exposure of fetal endothelial cells to a myo-inosit
ol concentration which approximated to a high fetal as opposed to a lo
w adult blood level, cell myo-inositol levels doubled and high-affinit
y transport underwent down-regulation.