Kiningham K, Bi X, Kasarskis E J
Department of Neurology, Veterans Administration Medical Center, Lexington, KY, USA.
Neurochem Int. 1995 Jul;27(1):105-9. doi: 10.1016/0197-0186(94)00172-q.
The immunocytochemical localization of the metallothioneins (MT), a family of low molecular weight, heavy metal binding proteins, was investigated in rat and human spinal cord using two distinct antibodies. Polyclonal antibodies were raised to the peptide (SCKCKECKCTS), an epitope common to all human MT isoforms. The monoclonal antibody, E9 which recognizes a different epitope of the MT protein, was also used. Similar results were obtained with either antibody. Both antibodies recognized authentic MT from rabbit liver and horse kidney on Western blots. Immunoreactivity was localized primarily in the cytoplasm of motoneurons in the anterior horn of cervical and lumbar cord. However nuclear staining was present in some motoneurons in both rat and human cord. Intense nuclear, as well as cytoplasmic staining was seen in astrocytes. Capillary endothelia, ependymal cells, arachnoid and pia were also positive for MT.
利用两种不同的抗体,对大鼠和人类脊髓中金属硫蛋白(MT,一种低分子量的重金属结合蛋白家族)进行了免疫细胞化学定位研究。针对肽段(SCKCKECKCTS)制备了多克隆抗体,该肽段是所有人类MT异构体共有的一个表位。还使用了识别MT蛋白不同表位的单克隆抗体E9。两种抗体均获得了相似的结果。在蛋白质免疫印迹法中,两种抗体均识别来自兔肝和马肾的真实MT。免疫反应性主要定位于颈髓和腰髓前角运动神经元的细胞质中。然而,在大鼠和人类脊髓的一些运动神经元中也存在核染色。在星形胶质细胞中可见强烈的核染色以及细胞质染色。毛细血管内皮、室管膜细胞、蛛网膜和软脑膜MT也呈阳性。