Slices of rat cerebral cortex incubated aerobically at 37 degrees C in Krebs-Ringer-bicarbonate solution accumulated 3,4-L-dihydroxyphenylalanine (L-DOPA) against its concentration gradient. With 1 mM L-DOPA in the medium, tissue-water/medium concentration ratios of about 6 : 1 are reached, which are modified by the presence of other amino acids in the medium. 2. Kinetic analysis suggested that L-DOPA influx into brain cells occurred by at least two saturable processes, which show apparent Km values in the range of 10(-3) M and 10(-5) M, respectively. 3. Prior incubation of the slices in Na+-free (choline-containing) medium at 37 degrees C depressed their subsequent uptake of L-DOPA in normal Na+-containing medium; this inhibition did not appear when the preincubation was carried out at 0-4 degrees C. Besides this effect of preincubation, most of L-DOPA influx into brain slices was independent of the actual concentration of Na+ in the medium; the two saturable processes described in this article behaved similarly in this respect. 4. Most of L-DOPA uptake by the high-Km process is mediated by an agency that resembles the Na+-independent L system described in Ehrlich cells (Oxender, D. L. and Christensen, H. N. (1963) J. Biol. Chem. 238, 2686-2699), both in its specificity and in its participation in exchange phenomena. A lesser component of uptake by a type A mediation is also suggested as contributing to the high-Km process . 5. The kinetic and specificity properties of the low-Km process of L-DOPA uptake suggest a similarity between its mediation and that of the high-affinity systems for L-tyrosine and L-tryptophan found in brain tissue preparations (Belin, M. F. and Pujol, J. F. (1973) Experientia 29, 411-413; Bauman, A., Bourgoin, S., Benda, P., Glowinski, J. and Hamon, M. (1971 Brain Res. 66, 253-263).
摘要
将大鼠大脑皮层切片在37℃下于 Krebs - 林格 - 碳酸氢盐溶液中进行需氧孵育时,其会逆着浓度梯度积累3,4 - L - 二羟基苯丙氨酸(L - DOPA)。当培养基中含有1 mM L - DOPA时,组织水/培养基浓度比可达约6 : 1,且该比值会因培养基中其他氨基酸的存在而改变。2. 动力学分析表明,L - DOPA流入脑细胞至少通过两个可饱和过程发生,这两个过程的表观 Km 值分别在10⁻³ M 和10⁻⁵ M 范围内。3. 将切片先在37℃的无钠(含胆碱)培养基中孵育,会降低其随后在正常含钠培养基中对L - DOPA的摄取;而当预孵育在0 - 4℃进行时,这种抑制作用不出现。除了预孵育的这种影响外,大部分L - DOPA流入脑切片的过程与培养基中实际的钠浓度无关;本文所述的两个可饱和过程在这方面表现相似。4. 高 Km 过程对L - DOPA的摄取大部分是由一种类似于 Ehrlich 细胞中描述的钠非依赖性 L 系统介导的(Oxender, D. L. 和 Christensen, H. N. (1963) J. Biol. Chem. 238, 2686 - 2699),在特异性及其参与交换现象方面均如此。还表明一种 A 型介导的摄取较小部分也对高 Km 过程有贡献。5. L - DOPA摄取的低 Km 过程的动力学和特异性特性表明,其介导方式与脑组织制剂中发现的L - 酪氨酸和L - 色氨酸的高亲和力系统的介导方式相似(Belin, M. F. 和 Pujol, J. F. (1973) Experientia 29, 411 - 413;Bauman, A., Bourgoin, S., Benda, P., Glowinski, J. 和 Hamon, M. (1971) Brain Res. 66, 253 - 263)。