BIOCHEMISTRY OF LEWY BODIES AND GELSOLIN AMYLOID

路易小体(Lbs)是一种独特的神经内胞内包涵体。 发病机制不明。在所有帕金森氏症病例中都发现了它们。 (PD)和弥漫性路易体病(DLBD)(也称为路易体 阿尔茨海默病的变种)。此外，偶尔还会发现lb。 在其他一些病理过程中。LBS的组成由直接 生化方法及其在这些疾病过程中的作用 未知。 最近，我们已经证明了提纯的淀粉样蛋白提取的抗体 来自家族性淀粉样变性，芬兰型(FAF)免疫反应与LBS。这 免疫反应性是特异的，因为它被纯化的淀粉样蛋白吸收。 蛋白质，不受泛素等其他抗原的影响， 神经丝蛋白和微管蛋白 免疫组织化学方法在LBS中的表达。FAF淀粉样蛋白一直是 发现是一种异常降解产物，从第173位开始， 明胶蛋白，一种肌动蛋白调节蛋白。氨基酸序列显示 N-末端和15位天冬酰胺的异质性 天冬氨酸的替代品。在DNA水平上，鸟嘌呤对腺嘌呤 与人明胶蛋白的654位核苷酸相对应的转变 到目前为止，在所有检测的Faf患者中都发现了这种病毒。市面上可买到的反- 凝胶素抗体上升到分子的羧基末端(离开 来自淀粉样变性区域)不要对LBS进行免疫染色。这引发了 LBS还可能含有一种异常的明胶蛋白片段，即 更靠近分子的氨基末端。我们计划：(1)执行 DLBD、阿尔茨海默病和帕金森病的免疫组织化学研究 有抗Faf抗体的疾病病例，以及有抗体的 不同区域的完整明胶、片段和合成肽 明胶分子，以评估免疫反应性和表位模式 作图；(2)表征纯化的LBS的组成 用蛋白质印迹和氨基酸直接测序的方法制备 对明胶蛋白和/或其片段进行测序；(3)研究 从DLBD、PD、AD和对照脑脊液中分离的明胶蛋白的活性 有无异常；(4)克隆家族性明胶蛋白基因 帕金森病患者和筛查任何变异；(5)脑部检查 帕金森病，DLBD，其他神经退行性疾病患者的脊髓液， 对照和AD检查是否存在任何异常的明胶蛋白片段 可以作为一种诊断测试。总而言之，这些研究将扩大 关于LBS的知识和明胶在其形成中所起的作用。

Lewy Bodies (LBs) are distinctive intraneural cytoplasmic inclusions of unknown pathogenesis. They are found in all cases of Parkinson's Disease (PD), and diffuse Lewy body disease (DLBD) (also known as the Lewy body variant of Alzheimer's disease). In addition, LBs are occasionally found in a few other pathological processes. The composition of LBs by direct biochemical methods and their role in these diseases processes remains unknown. Recently we have shown that antibodies raised to purified amyloid extracted from familial amyloidosis, Finnish type (FAF) immunoreact with LBs. This immunoreactivity is specific, as it was absorbed by the purified amyloid proteins and is unaffected by other antigens such as ubiquitin, neurofilament proteins and tubulin which have been shown by immunohistochemical methods to be present in LBs. FAF amyloid has been found to be an aberrant degradation product, starting at position 173, of gelsolin, an actin modulating protein. The amino acid sequence shows heterogeneity at the N-terminus and at position 15, where asparagine substitutes for aspartic acid. At the DNA level, a guanine to adenine transition corresponding to nucleotide 654 of the human gelsolin has been found in all FAF patients tested so far. Commercially available anti- gelsolin antibodies raised to the carboxy-terminus of the molecule (away from the amyloidogenic region) do not immunostain LBs. This raises the possibility that LBs also contain an abnormal fragment of gelsolin that is closer to the amino terminal of the molecule. We plan: (1) To perform an immunohistochemical survey of DLBD, Alzheimer's disease, and Parkinson's disease cases with the anti-FAF antibody, as well as with antibodies to intact gelsolin, fragments and synthetic peptides of different regions of the gelsolin molecule, to evaluate patterns of immunoreactivity and epitope mapping; (2) To characterize the composition of the purified LBs preparation by using Western blotting and direct amino acid sequencing to sequence gelsolin and/or its fragments; (3) To investigate the biological activity of gelsolin isolated from DLBD, PD, AD, and control CSF to see if there are any abnormalities; (4) To clone the gelsolin gene from familial PD patients and screen for any variations; (5) To investigate cerebral spinal fluid from patients with PD, DLBD, other neurodegenerative disorder, controls and AD for the presence of any abnormal gelsolin fragments that could be used as a diagnostic test. Together these studies will expand knowledge about LBs and the role gelsolin plays in their formation.