Harada Yuka, Takayama Fumiko, Tanabe Kazunari, Ni Junjun, Hayashi Yoshinori, Yamamoto Kenji, Wu Zhou, Nakanishi Hiroshi
Department of Aging Science and Pharmacology, Faculty of Dental Science, Kyushu University, Fukuoka, 812-8582, Japan.
Tanabe Preserved Dentistry, Fukuoka, 813-0041, Japan.
Cell Mol Neurobiol. 2017 Apr;37(3):437-443. doi: 10.1007/s10571-016-0376-x. Epub 2016 Apr 26.
Cathepsin E (CatE), an aspartic protease, has a limited distribution in certain cell types such as gastric cells. CatE is not detectable in the normal brain, whereas it is increasingly expressed in damaged neurons and activated microglia of the pathological brain. Neurons expressing high levels of CatE showed apparent morphological changes, including a marked shrinkage of the cytoplasmic region and beading of neurites, suggesting neuronal damage. The intracellular level of CatE in neurons is strictly regulated at both transcriptional and translational levels. Although the up-regulation of CatE may cause pathological changes in neurons, little information is available about the precise outcome of the increased expression of CatE in neurons. In this study, we have attempted to clarify the outcome of up-regulated CatE gene expression in neurons using the P19 cell neuronal differentiation after the overexpression of CatE. We unexpectedly found that the overexpression of CatE interfered with neuronal differentiation of P19 cells through an impairment of cell aggregate formation. Pepstatin A, an aspartic protease inhibitor, restored the impaired cell aggregation of P19/CatE cells. The small number of P19 cells differentiated into neurons had abnormal morphology characterized by their fusiform cell bodies with short processes. Furthermore, CatE proteolytically cleaved the extracellular domain of N-cadherin. These observations suggest that the overexpression of CatE interferes with neuronal differentiation of P19 cells through an impairment of cell aggregate formation, possibly through proteolytic degradation of N-cadherin.
组织蛋白酶E(CatE)是一种天冬氨酸蛋白酶,在某些细胞类型(如胃细胞)中分布有限。在正常大脑中检测不到CatE,而在病理性大脑中受损的神经元和活化的小胶质细胞中其表达却日益增加。表达高水平CatE的神经元显示出明显的形态变化,包括细胞质区域明显收缩和神经突串珠化,提示神经元损伤。神经元中CatE的细胞内水平在转录和翻译水平上都受到严格调控。虽然CatE的上调可能导致神经元发生病理变化,但关于CatE在神经元中表达增加的确切结果却知之甚少。在本研究中,我们试图通过在CatE过表达后利用P19细胞神经元分化来阐明神经元中CatE基因表达上调的结果。我们意外地发现,CatE的过表达通过损害细胞聚集体形成而干扰了P19细胞的神经元分化。天冬氨酸蛋白酶抑制剂胃蛋白酶抑制剂A恢复了P19/CatE细胞受损的细胞聚集。少数分化为神经元的P19细胞具有异常形态,其特征是梭形细胞体和短突起。此外,CatE蛋白水解切割N-钙黏蛋白的细胞外结构域。这些观察结果表明,CatE的过表达通过损害细胞聚集体形成,可能通过N-钙黏蛋白的蛋白水解降解来干扰P19细胞的神经元分化。