Cellular and Molecular Neuroscience, National Brain Research Centre, Manesar, Gurgaon 122 050, India.
J Biol Chem. 2013 Mar 29;288(13):9482-90. doi: 10.1074/jbc.M112.416180. Epub 2013 Feb 13.
Lafora disease (LD) is a teenage-onset inherited progressive myoclonus epilepsy characterized by the accumulations of intracellular inclusions called Lafora bodies and caused by mutations in protein phosphatase laforin or ubiquitin ligase malin. But how the loss of function of either laforin or malin causes disease pathogenesis is poorly understood. Recently, neuronatin was identified as a novel substrate of malin that regulates glycogen synthesis. Here we demonstrate that the level of neuronatin is significantly up-regulated in the skin biopsy sample of LD patients having mutations in both malin and laforin. Neuronatin is highly expressed in human fetal brain with gradual decrease in expression in developing and adult brain. However, in adult brain, neuronatin is predominantly expressed in parvalbumin-positive GABAergic interneurons and localized in their processes. The level of neuronatin is increased and accumulated as insoluble aggregates in the cortical area of LD brain biopsy samples, and there is also a dramatic loss of parvalbumin-positive GABAergic interneurons. Ectopic expression of neuronatin in cultured neuronal cells results in increased intracellular Ca(2+), endoplasmic reticulum stress, proteasomal dysfunction, and cell death that can be partially rescued by malin. These findings suggest that the neuronatin-induced aberrant Ca(2+) signaling and endoplasmic reticulum stress might underlie LD pathogenesis.
拉佛拉病(LD)是一种青少年起病的遗传性进行性肌阵挛性癫痫,其特征是细胞内包涵体的积累,称为拉佛拉体,由蛋白磷酸酶 laforin 或泛素连接酶 malin 的突变引起。但是,无论是 laforin 还是 malin 的功能丧失如何导致疾病的发病机制还知之甚少。最近,神经元蛋白被确定为 malin 的一种新底物,可调节糖原合成。在这里,我们证明在具有 malin 和 laforin 突变的 LD 患者的皮肤活检样本中,神经元蛋白的水平显著上调。神经元蛋白在人类胎脑中高度表达,在发育和成年脑中表达逐渐减少。然而,在成年脑中,神经元蛋白主要表达在 parvalbumin 阳性 GABA 能中间神经元中,并定位于其过程中。神经元蛋白的水平在 LD 脑活检样本的皮质区域中增加并积累为不溶性聚集体,并且也存在 parvalbumin 阳性 GABA 能中间神经元的显著丧失。神经元蛋白在培养的神经元细胞中的异位表达导致细胞内 Ca(2+)增加、内质网应激、蛋白酶体功能障碍和细胞死亡,而 malin 可部分挽救这些变化。这些发现表明,神经元蛋白诱导的异常 Ca(2+)信号和内质网应激可能是 LD 发病机制的基础。