Differential screening of mutated SOD1 transgenic mice reveals early up-regulation of a fast axonal transport component in spinal cord motor neurons
Differential screening of mutated SOD1 transgenic mice reveals early up-regulation of a fast axonal transport component in spinal cord motor neurons
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DOI：
10.1006/nbdi.2000.0292
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发表时间：
2000-08-01
影响因子：
6.1
通讯作者：
Loeffler, JP
中科院分区：
文献类型：
--
作者：
Dupuis, L;de Tapia, M;Loeffler, JP
在本研究中，我们分析了家族性肌萎缩侧索硬化症(FALS)运动神经元变性的分子机制。为此，我们使用了一个表达铜/锌超氧化物歧化酶(SOD1)基因的转基因小鼠模型，其中Gly(86)到Arg(G86R)突变与在人类FAL子集中发现的突变相同。采用一种优化的抑制消减杂交方法，通过序列分析，确定了G86R小鼠腰髓中一个在无症状时特异性上调的cDNAKIF3相关蛋白(KAP3)，它是一种快速轴突运输的调节因子。RT-PCR分析表明，KAP3的诱导是在轴突变性之前很久就发生的早期事件。免疫组织化学研究进一步表明，KAP3蛋白主要聚集在脊髓腹侧的大型运动神经元中。我们进一步证明，KAP3的上调不依赖于Kinesin II复合体的其他成分的任何变化。然而，由于普遍存在的KIF1A马达被上调，我们的结果显示在FALS病理过程中快速轴突运输机制的早期和复杂的重排。(C)2000年学术出版社。
In the present study we analyze the molecular mechanisms underlying motor neuron degeneration in familial amyotrophic lateral sclerosis (FALS). For this, we used a transgenic mouse model expressing the Cu/Zn superoxide dismutase (SOD1) gene with a Gly(86) to Arg (G86R) mutation equivalent to that found in a subset of human FALS. Using an optimized suppression subtractive hybridization method, a cDNA specifically up-regulated during the asymptomatic phase in the lumbar spinal cord of G86R mice was identified by sequence analysis as the KIF3-associated protein (KAP3), a regulator of fast axonal transport. RT-PCR analysis revealed that KAP3 induction was an early event arising long before axonal degeneration. Immunohistochemical studies further revealed that KAP3 protein predominantly accumulates in large motor neurons of the ventral spinal cord. We further demonstrated that KAP3 up-regulation occurs independent of any change in the other components of the kinesin II complex. However, since the ubiquitous KIF1A motor is up-regulated, our results show an early and complex rearrangement of the fast axonal transport machinery in the course of FALS pathology. (C) 2000 Academic Press.