Wisniewski K E, Kida E, Gordon-Majszak W, Saitoh T
New York State Office of Mental Retardation and Developmental Disabilities, Department of Pathological Neurobiology, Staten Island 10314.
Neurosci Lett. 1990 Nov 27;120(1):94-6. doi: 10.1016/0304-3940(90)90176-a.
Our previous study disclosed strong immunostaining of brain tissue in neuronal ceroid lipofuscinosis (NCL) with antibodies against amyloid beta-protein and the presence of 31-kDa polypeptide in the storage material. In the present study, we investigated the immunoreactivity of the NCL brain tissue with anti-serum (anti-GID) raised against a synthetic peptide, based on the amyloid beta-protein precursor, with the 175-186 amino acid sequence. Immunocytochemistry was performed on autopsy brain material collected from 15 NCL cases, and from 8 age-matched normal controls. The results showed strong immunoreactivity of nerve cells in the NCL cases, which according to densitometry was 5 times more intense than in the control group (P less than 0.0001 by Student's t-test). Western blot analysis revealed that in protein fractions of NCL brain homogenates anti-GID recognized the protein band of 35 kDa. Thus our present and previously performed studies supplied for the first time data pointing to abnormal processing of amyloid beta-protein precursor in NCL. Moreover, the accumulation of both 31- and 35-kDa polypeptides that was demonstrated provides further support for postulated defective protein metabolism in this disorder.
我们之前的研究发现，针对β-淀粉样蛋白的抗体对神经元蜡样脂褐质沉积症（NCL）脑组织有强烈的免疫染色，且储存物质中存在31 kDa多肽。在本研究中，我们基于β-淀粉样蛋白前体的175 - 186氨基酸序列合成了一种肽，并以此制备了抗血清（抗-GID），研究了NCL脑组织与该抗血清的免疫反应性。对从15例NCL病例及8例年龄匹配的正常对照者尸检获取的脑材料进行了免疫细胞化学检测。结果显示，NCL病例中的神经细胞有强烈的免疫反应性，根据光密度测定，其强度比对照组高5倍（学生t检验，P < 0.0001）。蛋白质印迹分析表明，在NCL脑匀浆的蛋白质组分中，抗-GID识别出了35 kDa的蛋白条带。因此，我们目前及之前进行的研究首次提供了指向NCL中β-淀粉样蛋白前体异常加工的数据。此外，所证实的31 kDa和35 kDa多肽的积累为该疾病中假定的蛋白质代谢缺陷提供了进一步支持。
