Heinonen O, Kyttälä A, Lehmus E, Paunio T, Peltonen L, Jalanko A
Department of Human Molecular Genetics, National Public Health Institute, Helsinki, 00300, Finland.
Mol Genet Metab. 2000 Feb;69(2):123-9. doi: 10.1006/mgme.2000.2961.
Infantile neuronal ceroid lipofuscinosis (INCL) is a severe neurodegenerative disorder in childhood that is caused by mutations in the gene encoding lysosomal palmitoyl protein thioesterase (PPT). INCL is characterized by massive and selective loss of cortical neurons. Here we have analyzed the intracellular processing and localization of adenovirus-mediated PPT in mouse primary neurons and NGF-induced PC-12 cells. The neuronal processing of PPT was found to be similar to that observed in peripheral cells, and a significant amount of the PPT enzyme was secreted in the primary neurons. Immunofluorescence analysis of the neuronal cells infected with wild-type PPT showed a granular staining pattern in the cell soma and neuronal shafts. Interestingly, PPT was also found in the synaptic ends of the neuronal cells and the staining pattern of the enzyme colocalized to a significant extent with the synaptic markers SV2 and synaptophysin. These in vitro data correspond with the distribution of endogeneous PPT in mouse brain and suggest that PPT may not solely be a lysosomal hydrolase. The specific targeting of PPT into the neuritic shafts and nerve terminals indicates that PPT may be associated with the maintenance of synaptic function. Furthermore, since a substantial amount of PPT is secreted by neurons, it is tempting to speculate that the enzyme could also have an extracellular substrate.
婴儿神经元蜡样脂褐质沉积症(INCL)是一种严重的儿童神经退行性疾病,由编码溶酶体棕榈酰蛋白硫酯酶(PPT)的基因突变引起。INCL的特征是皮质神经元大量选择性丧失。在此,我们分析了腺病毒介导的PPT在小鼠原代神经元和神经生长因子(NGF)诱导的PC-12细胞中的细胞内加工和定位情况。发现PPT在神经元中的加工过程与在外周细胞中观察到的相似,并且在原代神经元中分泌出大量的PPT酶。对感染野生型PPT的神经元细胞进行免疫荧光分析,结果显示在细胞体和神经轴突中呈现颗粒状染色模式。有趣的是,在神经元细胞的突触末端也发现了PPT,并且该酶的染色模式在很大程度上与突触标记物SV2和突触素共定位。这些体外数据与内源性PPT在小鼠脑中的分布情况相符,表明PPT可能不仅仅是一种溶酶体水解酶。PPT特异性靶向神经轴突和神经末梢表明,PPT可能与突触功能的维持有关。此外,由于神经元分泌大量的PPT,因此很容易推测该酶可能也有细胞外底物。