Francis A, Rivett A J, Roth J A
Brain Res. 1983 Mar 14;263(1):89-95. doi: 10.1016/0006-8993(83)91203-9.
Rat brain contains 3 forms of enolase, a neuron-specific form (NSE), a hybrid form, and a non-neuronal form (NNE) which were separated by DEAE-cellulose column chromatography. The enolase activity corresponding to each form of the enzyme eluted from the columns was determined spectrophotometrically. Using this assay procedure, an activity ratio (%NNE/%NSE) was calculated for cerebellum, brainstem, sciatic nerve, adrenals and liver. The results indicated excellent agreement between this enzymatically determined ratio and published values of a similar ratio (NNE/NSE) determined by radioimmunoassay for enzyme protein. Following in vivo destruction of neurons by intracerebral injection of the selective neurotoxin, kainic acid, there was a significant decrease in the activity of NSE and hybrid enolase (neuronal forms) and no change in the activity of NNE (glial form). These data indicate that separation and measurement of enolase species is useful to determine levels of these species in normal tissue and to estimate neuronal damage biochemically in brain lesions.
大鼠脑含有3种形式的烯醇化酶,一种神经元特异性形式(NSE)、一种混合形式和一种非神经元形式(NNE),它们通过DEAE - 纤维素柱色谱法分离。从柱上洗脱的每种酶形式对应的烯醇化酶活性通过分光光度法测定。使用该测定程序,计算了小脑、脑干、坐骨神经、肾上腺和肝脏的活性比(%NNE/%NSE)。结果表明,该酶法测定的比率与通过放射免疫法测定的酶蛋白类似比率(NNE/NSE)的已发表值之间具有极好的一致性。在脑内注射选择性神经毒素红藻氨酸对神经元进行体内破坏后,NSE和混合烯醇化酶(神经元形式)的活性显著降低,而NNE(神经胶质形式)的活性没有变化。这些数据表明,烯醇化酶种类的分离和测量有助于确定正常组织中这些种类的水平,并在脑损伤中通过生化方法估计神经元损伤。