Developmental modulation of blood-brain-barrier glucose transport in the rabbit

Abstract

Blood-brain barrier (BBB) glucose transport rates were measured using the intracarotid injection method in newborn, 14-day-old suckling, 28-day-old weanling and adult rabbits, and compared with membrane transporter density. Light microscope immunochemistry confirmed the presence of the GLUT1 glucose transporter isoform in these rabbits. Quantitative electron microscopic immunogold analyses of GLUT1-immunoreactive sites per micrometer of capillary membrane indicated GLUT1 density increased with age, and correlated with in vivo measurements of V max. Maximal transport velocities ( V max) of glucose transfer (an indicator of the activity and relative number of transporter proteins) increased significantly ( P=0.05) with age: in neonates V max = 0.61 μmol · min. −1 · g −1, in sucklings V max = 0.68 μmol. · min −1 · g −1, in weanlings V max = 0.88 μmol · min −1 · g −1, and in adults V max = 1.01 μmol · min −1 g −1. Cerebral blood flow (CBF) rates, increased with age from 0.19 and 0.26 ml · min −1 · g −1 in neonates and sucklings to 0.51 (weanlings) and 0.70 (adults) ml · min −1 · g −. Non-linear regression analyses indicated the half-saturation constant ( K m) for glucose transport ranged from 13 mM in adult rabbits to 19 mM in 14-day-old sucklings: differences in K m were not significant. Age-related changes in the Permeability-Surface Area product ( PS ± S.E.) of both water and glucose were also seen. At all ages studied, the diffusion component ( K d) of glucose uptake was not distinguishable from zero. We conclude developmental up-regulation of the rabbit BBB glucose transporter is characterized by no changes in transporter affinity, and provide the first demonstration of increased membrane transporter proteins correlating with an age-related increase (65%) in glucose transporter maximal velocity.