Ofori S, Bretton C, Hof P, Schorderet M
J Neurochem. 1986 Oct;47(4):1199-206. doi: 10.1111/j.1471-4159.1986.tb00740.x.
The basal catecholamine content of rabbit retina was determined by liquid chromatography with electrochemical detection (LC-EC) and 3,4-dihydroxyphenylethylamine (dopamine, DA) found to be the major catecholamine. The immediate DA precursor, 3,4-dihydroxyphenylalanine (L-DOPA), and the metabolite, 3,4-dihydroxyphenylacetic acid (DOPAC), were also detected at about 2.8% and 17% of DA levels, respectively. When added exogenously, L-tyrosine did not increase the rate of DA synthesis over the basal level. In contrast, exogenous L-DOPA led to a 3.5-fold increase in DA, and to a 20-fold increase in DOPAC content. The monoamine oxidase inhibitors pargyline and (-)-deprenyl differentially affected the degradation of DA, since 100 microM pargyline was apparently more effective than 100 microM (-)-deprenyl. Reserpine and (+/-)-amphetamine each induced a Ca2+-independent decrease of DA stores. The separate actions of reserpine and (+/-)-amphetamine in lowering tissue DA levels were additive, suggesting two separate pools of DA available for release from presynaptic stores. The present study demonstrates that the LC-EC technique may be used to investigate the modulation of the synthesis and release of retinal DA in vitro, without the prior uptake of radiolabelled transmitter.
采用液相色谱-电化学检测法(LC-EC)测定了兔视网膜中基础儿茶酚胺的含量,发现3,4-二羟基苯乙胺(多巴胺,DA)是主要的儿茶酚胺。DA的直接前体3,4-二羟基苯丙氨酸(L-多巴)及其代谢产物3,4-二羟基苯乙酸(DOPAC)的含量也分别检测到,约为DA水平的2.8%和17%。外源性添加L-酪氨酸时,DA合成速率并未超过基础水平。相反,外源性L-多巴使DA增加3.5倍,DOPAC含量增加20倍。单胺氧化酶抑制剂优降宁和(-)-司来吉兰对DA降解的影响不同,因为100μM优降宁的作用明显强于100μM(-)-司来吉兰。利血平和(±)-苯丙胺均引起了DA储存的Ca2+非依赖性减少。利血平和(±)-苯丙胺降低组织DA水平的作用是相加的,提示有两个可从突触前储存中释放的独立DA池。本研究表明,LC-EC技术可用于体外研究视网膜DA合成与释放的调节,而无需事先摄取放射性标记递质。