Geiger J D, Nagy J I
J Neurosci. 1986 Sep;6(9):2707-14. doi: 10.1523/JNEUROSCI.06-09-02707.1986.
The activity of adenosine deaminase (ADA) was measured in 62 discrete regions of the CNS, and in some autonomic and sensory ganglia, peripheral nerves, and peripheral tissues of the rat using an automated high-pressure liquid chromatography (HPLC) method. The formation of inosine and hypoxanthine as a measure of ADA activity in homogenates of brain was optimal at pH 7.0, linear for up to 60 min at 37 degrees C using 500 microM adenosine as substrate, and linear with protein concentrations ranging from 0.05 to 0.8 mg. The Km and Vmax values for ADA activity in homogenates of whole brain were 47 microM and 107 nmol/mg protein/30 min, respectively. Among the CNS regions examined, the highest activity was found in posterior hypothalamic magnocellular nuclei and the lowest in hippocampus. In general, spinal cord contained relatively low levels of ADA activity, with that in dorsal cord approximately 40% higher than ventral cord. In the periphery, parasympathetic ganglia contained higher levels of ADA than sensory ganglia and brain. Most peripheral tissues--including adrenal gland, lung, liver, and anterior and posterior pituitary--exhibited activity comparable to levels in the posterior hypothalamus. ADA activity in thymus was about 10 times higher than any other tissue examined. The uneven distribution of ADA activity in the rat CNS corresponds well with the immunohistochemical localization of this enzyme in discrete neural systems of this species. Structures that contain high ADA activity exhibit intense ADA immunostaining of neuronal perikarya and/or fibers.(ABSTRACT TRUNCATED AT 250 WORDS)
采用自动高压液相色谱(HPLC)法,测定了大鼠中枢神经系统(CNS)的62个离散区域、部分自主神经节和感觉神经节、外周神经及外周组织中腺苷脱氨酶(ADA)的活性。以次黄苷和次黄嘌呤的生成量作为脑匀浆中ADA活性的衡量指标,在pH 7.0时最为适宜,以500微摩尔/升腺苷为底物,在37摄氏度下反应60分钟内呈线性关系,且与蛋白质浓度在0.05至0.8毫克范围内呈线性关系。全脑匀浆中ADA活性的米氏常数(Km)和最大反应速度(Vmax)值分别为47微摩尔/升和107纳摩尔/毫克蛋白质/30分钟。在所检测的中枢神经系统区域中,下丘脑后部大细胞神经核的活性最高,海马体的活性最低。一般来说,脊髓中ADA活性水平相对较低,背侧脊髓中的活性比腹侧脊髓高约40%。在周围组织中,副交感神经节中的ADA水平高于感觉神经节和脑。大多数外周组织,包括肾上腺、肺、肝以及垂体前叶和后叶,其活性与下丘脑后部的水平相当。胸腺中的ADA活性比所检测的任何其他组织高约10倍。大鼠中枢神经系统中ADA活性的不均匀分布与该酶在该物种离散神经系统中的免疫组织化学定位非常吻合。含有高ADA活性的结构对神经元胞体和/或纤维呈现强烈的ADA免疫染色。(摘要截短于250字)