Kusunoki S, Chiba A, Hirabayashi Y, Irie F, Kotani M, Kawashima I, Tai T, Nagai Y
Department of Neurology, Faculty of Medicine, University of Tokyo, Japan.
Brain Res. 1993 Sep 24;623(1):83-8. doi: 10.1016/0006-8993(93)90013-d.
We have established a monoclonal antibody, GGR41, specific for a new class of minor gangliosides, such as GQ1b alpha and GT1a alpha, by immunizing mice with a GQ1b-rich ganglioside fraction extracted from bovine brain. Each of those minor gangliosides has been reported to be one of the cholinergic-specific gangliosides (Chol-1). Careful examination of binding specificity of the antibody by both an enzyme-linked immunosorbent assay and immunostaining on thin-layer chromatograms showed that the antibody recognizes three sialyl residues separately attaching to the gangliotetraosyl backbone structure. Immunohistochemical analysis revealed that GGR41 immunostained lamina I and III of dorsal horn and lateral horn of human thoracic cord but motor neurons were not immunostained. Except for negative staining of motor neurons, this distribution is similar to the distribution pattern of staining as reported in rats and humans using a polyclonal antibody against Chol-1. Thus, the antibody obtained in this study should be a useful reagent to study the function of a unique new class of the minor gangliosides.
我们通过用从牛脑中提取的富含GQ1b的神经节苷脂组分免疫小鼠,建立了一种针对新型次要神经节苷脂(如GQ1bα和GT1aα)的单克隆抗体GGR41。据报道,这些次要神经节苷脂中的每一种都是胆碱能特异性神经节苷脂(Chol-1)之一。通过酶联免疫吸附测定和薄层色谱免疫染色对抗体结合特异性进行仔细检查表明,该抗体识别分别连接到神经节四糖主链结构上的三个唾液酸残基。免疫组织化学分析显示,GGR41对人胸段脊髓背角和外侧角的I层和III层进行免疫染色,但运动神经元未被免疫染色。除运动神经元呈阴性染色外,这种分布与使用抗Chol-1多克隆抗体在大鼠和人类中报道的染色分布模式相似。因此,本研究中获得的抗体应该是研究一类独特新型次要神经节苷脂功能的有用试剂。