Differential expression of splice variant and wild-type parkin in sporadic Parkinson's disease
Differential expression of splice variant and wild-type parkin in sporadic Parkinson's disease
复制标题
DOI：
10.1007/s10048-005-0001-5
复制
发表时间：
2005-12-01
期刊：
影响因子：
2.2
通讯作者：
Zhao, Y
中科院分区：
文献类型：
--
作者：
Tan, EK;Shen, H;Zhao, Y
背景：细胞应激条件下parkin剪接的改变可导致基因表达的改变和蛋白质活性的改变。帕金在散发性帕金森病（PD）中的致病作用尚不清楚。目的：我们描述了一个帕金剪接变异体（SV）在黑质和白细胞的散发性PD患者。采用病例对照方法，我们研究了外显子4 SV（E4 SV）和野生型帕金蛋白在散发性PD患者和健康个体的白细胞中的表达。方法/结果：我们通过逆转录-聚合酶链反应（PCR）在散发性PD患者和对照组的黑质和白细胞中发现了一种parkin E4SV。外显子4（122 bp）缺失导致外显子3 - 5连接处的阅读移码和外显子3下游17 bp的终止密码子（tga）。翻译的截短蛋白与两个环指功能结构域的完全丧失有关。利用TaqMan实时PCR，探针位于外显子3 - 4或3 - 5的连接处，我们证明了与年龄、性别和种族匹配的对照组相比，散发性PD患者的白细胞中E4SV/野生型parkin比率过表达（p <0.0005）。多变量回归分析表明，在PD患者中，E4SV/野生型parkin表达的比率随年龄增加而增加，但在对照组中未观察到这一点（p <0.0005）。结论：E4 SV/野生型parkin基因在散发性PD中的相对表达高于健康对照。根据我们的观察，进一步的功能研究，以确定E4 SV在散发性PD患者的病理生理作用将是重要的。
Background: Altered splicing of parkin under cellular stress could lead to changes in gene expression and altered protein activity. The causative role of parkin in sporadic Parkinson's disease (PD) is unknown. Objectives: We described a parkin splice variant (SV) in the substantia nigra and leukocytes of sporadic PD patients. Using a case control methodology, we investigated the exon 4 SV (E4SV) and wild-type parkin expression in the leukocytes of sporadic PD patients and healthy individuals. Methods/Results: We identified a parkin E4SV in the substantia nigra and leukocytes of sporadic PD patients and controls by reverse transcriptase-polymerase chain reaction (PCR). The exon 4 (122 bp) deletion resulted in a reading frame shift over the junction of exons 3-5 and a stop codon (tga) 17 bp downstream from exon 3. The translated truncated protein was associated with a total loss of the two-RING finger functional domain. Utilizing TaqMan real-time PCR with probes located across the junction of exons 3-4 or 3-5, we demonstrated an over-expression of E4SV/wild-type parkin ratio in the leukocytes of sporadic PD patients compared to age-, gender-, and race-matched controls (p < 0.0005). A multivariate regression analysis demonstrated that the ratio of E4SV/wild-type parkin expression increased with age in PD patients, but this was not observed in the controls (p < 0.0005). Conclusion: The relative expression of E4SV/wild type parkin was increased in sporadic PD compared to healthy controls. Based on our observations, further functional studies to determine the pathophysiologic role of E4SV in sporadic PD patients will be of importance.