Equipe Différenciation Neuromusculaire, CNRS, UMR5239, Ecole Normale Supérieure-Lyon, 46 allée d'Italie, 69364, Lyon Cedex 07, France,
Cell Mol Life Sci. 2014 Sep;71(17):3363-79. doi: 10.1007/s00018-014-1566-7. Epub 2014 Feb 4.
Nesprin-1 is a core component of a protein complex connecting nuclei to cytoskeleton termed LINC (linker of nucleoskeleton and cytoskeleton). Nesprin-1 is anchored to the nuclear envelope by its C-terminal KASH domain, the disruption of which has been associated with neuronal and neuromuscular pathologies, including autosomal recessive cerebellar ataxia and Emery-Dreifuss muscular dystrophy. Here, we describe a new and unexpected role of Drosophila Nesprin-1, Msp-300, in neuromuscular junction. We show that larvae carrying a deletion of Msp-300 KASH domain (Msp-300 (∆KASH) ) present a locomotion defect suggestive of a myasthenia, and demonstrate the importance of muscle Msp-300 for this phenotype, using tissue-specific RNAi knock-down. We show that Msp-300 (∆KASH) mutants display abnormal neurotransmission at the larval neuromuscular junction, as well as an imbalance in postsynaptic glutamate receptor composition with a decreased percentage of GluRIIA-containing receptors. We could rescue Msp-300 (∆KASH) locomotion phenotypes by GluRIIA overexpression, suggesting that the locomotion impairment associated with the KASH domain deletion is due to a reduction in junctional GluRIIA. In summary, we found that Msp-300 controls GluRIIA density at the neuromuscular junction. Our results suggest that Drosophila is a valuable model for further deciphering how Nesprin-1 and LINC disruption may lead to neuronal and neuromuscular pathologies.
Nesprin-1 是一种将核与细胞骨架连接起来的蛋白质复合物(核骨架和细胞骨架的连接)的核心组成部分。Nesprin-1 通过其 C 端 KASH 结构域锚定在核膜上,该结构域的破坏与神经元和神经肌肉疾病有关,包括常染色体隐性小脑共济失调和 Emery-Dreifuss 肌营养不良症。在这里,我们描述了果蝇 Nesprin-1(Msp-300)在神经肌肉接点的一个新的、意想不到的作用。我们表明,携带 Msp-300 KASH 结构域缺失(Msp-300(∆KASH))的幼虫表现出运动缺陷,类似于重症肌无力,并使用组织特异性 RNAi 敲低证明了肌肉 Msp-300 对这种表型的重要性。我们表明,Msp-300(∆KASH)突变体在幼虫神经肌肉接点显示出异常的神经传递,以及突触后谷氨酸受体组成的不平衡,含有 GluRIIA 的受体百分比降低。我们可以通过 GluRIIA 过表达来挽救 Msp-300(∆KASH)的运动表型,这表明与 KASH 结构域缺失相关的运动障碍是由于连接点 GluRIIA 的减少所致。总之,我们发现 Msp-300 控制着神经肌肉接点处的 GluRIIA 密度。我们的结果表明,果蝇是进一步阐明 Nesprin-1 和 LINC 破坏如何导致神经元和神经肌肉疾病的有价值的模型。