Tayarani I, Lefauconnier J M, Roux F, Bourre J M
INSERM U26, Hôpital Fernand Widal, Paris, France.
J Cereb Blood Flow Metab. 1987 Oct;7(5):585-91. doi: 10.1038/jcbfm.1987.109.
Brain capillaries isolated from 2-month-old male and female Sprague-Dawley rats were used to study the transport of neutral amino acids. The uptake of alanine, leucine, and alpha-methylaminoisobutyric acid (MeAIB) was a linear function of time for the first minute of incubation. Based on these observations, an incubation time of 1 min was used to measure transport activities. The intracellular water volume of the isolated capillaries was 2.2 microliters/mg protein. This value was significantly lower (1.8 microliter/mg protein) when measured in the absence of sodium. L-Alanine, L-serine, and L-cysteine were taken up from the abluminal surface of brain capillaries by a sodium- and energy-dependent, carrier-mediated system. This uptake, for the most part, was not inhibited by MeAIB. System ASC (alanine-serine-cysteine) appeared to be of primary importance for the transport of these amino acids in isolated brain capillaries. The apparent Km and Vmax values for L-alanine uptake by ASC transport, based on the Hofstee plot presentation, were 1.3 mM and 0.975 nmol/microliter water content/min, respectively. The results also indicate that the transport of MeAIB and 2-aminobicyclo(2,2,1)heptane-2-carboxylic acid (BCH) was limited to the sodium-dependent system A (alanine) and the sodium-independent system L (leucine), respectively.
从2个月大的雄性和雌性Sprague-Dawley大鼠中分离出脑毛细血管,用于研究中性氨基酸的转运。在孵育的第一分钟,丙氨酸、亮氨酸和α-甲基氨基异丁酸(MeAIB)的摄取是时间的线性函数。基于这些观察结果,使用1分钟的孵育时间来测量转运活性。分离出的毛细血管的细胞内水体积为2.2微升/毫克蛋白质。在无钠条件下测量时,该值显著降低(1.8微升/毫克蛋白质)。L-丙氨酸、L-丝氨酸和L-半胱氨酸通过钠和能量依赖的载体介导系统从脑毛细血管的管腔表面摄取。这种摄取在很大程度上不受MeAIB的抑制。系统ASC(丙氨酸-丝氨酸-半胱氨酸)似乎对这些氨基酸在分离的脑毛细血管中的转运至关重要。基于霍夫斯泰(Hofstee)图表示法,ASC转运对L-丙氨酸摄取的表观Km和Vmax值分别为1.3 mM和0.975纳摩尔/微升水含量/分钟。结果还表明,MeAIB和2-氨基双环(2,2,1)庚烷-2-羧酸(BCH)的转运分别限于钠依赖的系统A(丙氨酸)和钠非依赖的系统L(亮氨酸)。