Department of Biology and Institute for Genome Sciences and Policy, Duke University, Durham, NC 27708, USA.
Hum Mol Genet. 2010 May 15;19(10):1883-96. doi: 10.1093/hmg/ddq064. Epub 2010 Feb 13.
Mutations in spastin are the most frequent cause of the neurodegenerative disease autosomal dominant-hereditary spastic paraplegia (AD-HSP). Drosophila melanogaster lacking spastin exhibit striking behavioral similarities to human patients suffering from AD-HSP, suggesting conservation of Spastin function between the species. Consistent with this, we show that exogenous expression of wild-type Drosophila or human spastin rescues behavioral and cellular defects in spastin null flies equivalently. This enabled us to generate genetically representative models of AD-HSP, which arises from dominant mutations in spastin rather than a complete loss of the gene. Flies co-expressing one copy of wild-type human spastin and one encoding the K388R catalytic domain mutation in the fly spastin null background, exhibit aberrant distal synapse morphology and microtubule distribution, similar to but less severe than spastin nulls. R388 or a separate nonsense mutation act dominantly and are furthermore sufficient to confer partial rescue, supporting in vitro evidence for additional, non-catalytic Spastin functions. Using this model, we tested the observation from human pedigrees that S44L and P45Q are trans-acting modifiers of mutations affecting the Spastin catalytic domain. As in humans, both L44 and Q45 are largely silent when heterozygous, but exacerbate mutant phenotypes when expressed in trans with R388. These transgenic 'AD-HSP' flies therefore provide a powerful and tractable model to enhance our understanding of the cellular and behavioral consequences of human spastin mutations and test hypotheses directly relevant to the human disease.
突变的痉挛蛋白是最常见的原因的神经退行性疾病常染色体显性遗传性痉挛性截瘫(AD-HSP)。果蝇缺乏痉挛蛋白表现出惊人的行为相似性,以人类患者患有 AD-HSP,表明在物种之间的 Spastin 功能的保守。与此一致,我们表明,外源性表达野生型果蝇或人类痉挛蛋白同样可以挽救痉挛蛋白 null 蝇的行为和细胞缺陷。这使我们能够产生 AD-HSP 的遗传代表性模型,其由痉挛蛋白中的显性突变引起,而不是完全缺失该基因。在果蝇痉挛蛋白 null 背景下共同表达一个野生型人类痉挛蛋白拷贝和一个编码果蝇痉挛蛋白的 K388R 催化结构域突变的拷贝的蝇,表现出异常的远端突触形态和微管分布,类似于但比痉挛蛋白 null 更为严重。R388 或单独的无意义突变表现为显性,并且足以提供部分挽救,支持体外证据表明 Spastin 具有其他非催化功能。使用这种模型,我们测试了来自人类家族的观察结果,即 S44L 和 P45Q 是影响 Spastin 催化结构域的突变的反式作用修饰因子。与人类一样,当杂合子时,L44 和 Q45 大部分是沉默的,但当与 R388 表达时,会加剧突变表型。这些转基因“AD-HSP”蝇因此提供了一个强大和可行的模型,以增强我们对人类痉挛蛋白突变的细胞和行为后果的理解,并直接测试与人类疾病相关的假设。