He Cui Zhen, Hays Arthur P
Department of Pathology, College of Physicians and Surgeons, Columbia University, PH 15th Stem, Room 124, 630 West 168th Street, New York, NY 10032, USA.
J Neurol Sci. 2004 Jan 15;217(1):47-54. doi: 10.1016/j.jns.2003.08.016.
We evaluated the expression of the type III intermediate filament (IF) protein, peripherin (PRP), in ubiquinated inclusions of motor neurons in amyotrophic lateral sclerosis (ALS). A previous study showed that overexpression of PRP in transgenic mice induces motor neuron disease with formation of PRP-containing inclusions before onset of symptoms [J. Cell Biol. 147 (3) (1999) 531]. To determine whether PRP inclusions occur in the human disease, we applied doublelabeling immunofluorescence to paraffin sections of the spinal cord obtained by autopsy of 40 ALS patients with sporadic disease and 39 controls. Inclusions that expressed immunoreactive ubiquitin and peripherin were recorded by video camera, and the sections were stained by hematoxylin and eosin (H&E) to define morphology. Lewy body-like inclusions (LBLIs) were seen in motor neuron perikarya of 9 of 40 ALS cases and none in controls; all LBLIs expressed peripherin. Skein-like inclusions (SLIs) were identified by ubiquitin, but did not express PRP with rare exceptions. Neither skein-like inclusions nor LBLIs expressed alpha B-crystallin, neurofilament protein (NF-L, NF-M and NF-H subunits), alpha-internexin, actin or alpha-synuclein. Immunoblot of the whole spinal cord exhibited a single 57-kDa band of peripherin in ALS patients and controls. Our data document the expression of peripherin in LBLIs, which may provide a clue to the pathogenesis of neurodegeneration in ALS.
我们评估了III型中间丝（IF）蛋白外周蛋白（PRP）在肌萎缩侧索硬化症（ALS）运动神经元泛素化包涵体中的表达。先前的一项研究表明，在转基因小鼠中过表达PRP会诱发运动神经元疾病，并在症状出现前形成含PRP的包涵体[《细胞生物学杂志》147(3)(1999)531]。为了确定PRP包涵体是否出现在人类疾病中，我们对40例散发性ALS患者和39例对照者尸检获得的脊髓石蜡切片进行了双重免疫荧光标记。用摄像机记录表达免疫反应性泛素和外周蛋白的包涵体，并用苏木精和伊红（H&E）对切片进行染色以确定形态。在40例ALS病例中有9例的运动神经元胞体中可见路易小体样包涵体（LBLIs），而对照者中未发现；所有LBLIs均表达外周蛋白。类丝状包涵体（SLIs）通过泛素鉴定，但除极少数例外均不表达PRP。类丝状包涵体和LBLIs均不表达αB-晶状体蛋白、神经丝蛋白（NF-L、NF-M和NF-H亚基）、α-中间丝蛋白、肌动蛋白或α-突触核蛋白。对整个脊髓进行免疫印迹分析，ALS患者和对照者中均出现一条单一的57 kDa外周蛋白条带。我们的数据证明了外周蛋白在LBLIs中的表达，这可能为ALS神经退行性变的发病机制提供线索。
