Wadhwani K C, Smith Q R, Rapoport S I
Laboratory of Neurosciences, National Institute on Aging, National Institutes of Health, Bethesda, Maryland 20892.
Am J Physiol. 1990 Jun;258(6 Pt 2):R1436-44. doi: 10.1152/ajpregu.1990.258.6.R1436.
L-Phenylalanine transport across the blood-nerve barrier was studied by perfusing Ringer solution containing trace amounts of L-[14C]phenylalanine and [3H]dextran (a vascular marker) through the hindlimb of an anesthetized rat. At the end of perfusion, a segment of sciatic nerve was removed, frozen, desheathed, and processed for radioactivity counting. The permeability-surface area product of the blood-nerve barrier (PABNB) to L-[14C]phenylalanine decreased from 58 +/- 9 X 10(-5) to 1.7 +/- 0.3 X 10(-5) (SE) ml.s-1.g wet wt-1, as the perfusate concentration of unlabeled L-phenylalanine was increased to 10 mM. D-Phenylalanine had less of an inhibitory effect on L-[14C]phenylalanine uptake than did L-phenylalanine. PABNB did not change in the presence of 50 mM alpha-(methylamino)isobutyric acid (MeAIB) in the perfusion medium, nor by replacing NaCl by Tris.HCl. However, addition of 50 mM 2-aminobicyclo(2,2,1)heptane-2-carboxylic acid (BCH) reduced PABNB to 2.1 +/- 0.3 X 10(-5) ml.s-1.g wet wt-1 (n = 4). PA at epiperineurial barrier (PAper) of L-[14C]phenylalanine at the epiperineurial barrier, determined from in situ incubation, was 1.4 +/- 0.2 X 10(-5) ml.s-1.g wet wt-1 (n = 6) and was unaffected by 10 mM L-phenylalanine concentration in the incubation medium. The results demonstrate for the first time carrier-mediated transport system for L-phenylalanine at the blood-nerve barrier of the rat peripheral nerve.
通过将含有微量L-[¹⁴C]苯丙氨酸和[³H]葡聚糖(一种血管标志物)的林格溶液灌注到麻醉大鼠的后肢,研究了L-苯丙氨酸跨血-神经屏障的转运。灌注结束时,取出一段坐骨神经,冷冻,剥去神经鞘,并进行放射性计数处理。随着未标记L-苯丙氨酸灌注液浓度增加至10 mM,血-神经屏障对L-[¹⁴C]苯丙氨酸的通透表面积乘积(PABNB)从58±9×10⁻⁵降至1.7±0.3×10⁻⁵(标准误)ml·s⁻¹·g湿重⁻¹。D-苯丙氨酸对L-[¹⁴C]苯丙氨酸摄取的抑制作用小于L-苯丙氨酸。灌注培养基中存在50 mM α-(甲基氨基)异丁酸(MeAIB)时,或用Tris·HCl替代NaCl时,PABNB均未改变。然而,添加50 mM 2-氨基双环(2,2,1)庚烷-2-羧酸(BCH)可使PABNB降至2.1±0.3×10⁻⁵ ml·s⁻¹·g湿重⁻¹(n = 4)。通过原位孵育测定的L-[¹⁴C]苯丙氨酸在神经外膜屏障处的通透系数(PAper)为1.4±0.2×10⁻⁵ ml·s⁻¹·g湿重⁻¹(n = 6),且不受孵育培养基中10 mM L-苯丙氨酸浓度的影响。结果首次证明了大鼠外周神经血-神经屏障处存在L-苯丙氨酸的载体介导转运系统。