IRCCS SDN-Foundation, Naples, Italy.
J Neurochem. 2014 Jun;129(6):1002-12. doi: 10.1111/jnc.12683. Epub 2014 Mar 24.
Phenylketonuria (PKU), if not detected and treated in newborns, causes severe neurological dysfunction and cognitive and behavioral deficiencies. Despite the biochemical characterization of PKU, the molecular mechanisms underlying PKU-associated brain dysfunction remain poorly understood. The aim of this study was to gain insights into the pathogenesis of this neurological damage by analyzing protein expression profiles in brain tissue of Black and Tan BRachyury-PahEnu2 mice (a mouse model of PKU). We compared the cerebral protein expression of homozygous PKU mice with that of their heterozygous counterparts using two-dimensional difference gel electrophoresis analysis, and identified 21 differentially expressed proteins, four of which were over-expressed and 17 under-expressed. An in silico bioinformatic approach indicated that protein under-expression was related to neuronal differentiation and dendritic growth, and to such neurological disorders as progressive motor neuropathy and movement disorders. Moreover, functional annotation analyses showed that some identified proteins were involved in oxidative metabolism. To further investigate the proteins involved in the neurological damage, we validated two of the proteins that were most strikingly under-expressed, namely, Syn2 and Dpysl2, which are involved in synaptic function and neurotransmission. We found that Glu2/3 and NR1 receptor subunits were over-expressed in PKU mouse brain. Our results indicate that differential expression of these proteins may be associated with the processes underlying PKU brain dysfunction, namely, decreased synaptic plasticity and impaired neurotransmission. We identified a set of proteins whose expression is affected by hyperphenylalaninemia. We think that phenylketonuria (PKU) brain dysfunction also depends on reduced Syn2 and Dpysl2 levels, increased Glu2/3 and NR1 levels, and decreased Pkm, Ckb, Pgam1 and Eno1 levels. These findings finally confirm that alteration in synaptic function, in transmission and in energy metabolism underlie brain damage provoked by hyperphenylalaninemias.
苯丙酮尿症(PKU)如果在新生儿中未被发现和治疗,会导致严重的神经功能障碍以及认知和行为缺陷。尽管已经对 PKU 进行了生化特征分析,但导致 PKU 相关脑功能障碍的分子机制仍知之甚少。本研究旨在通过分析 Black 和 Tan BRachyury-PahEnu2 小鼠(PKU 的一种小鼠模型)脑组织中的蛋白质表达谱,深入了解这种神经损伤的发病机制。我们使用二维差异凝胶电泳分析比较了纯合 PKU 小鼠和杂合 PKU 小鼠的大脑蛋白质表达,鉴定出 21 种差异表达的蛋白质,其中 4 种表达上调,17 种表达下调。计算机生物信息学方法表明,蛋白质表达下调与神经元分化和树突生长以及进行性运动神经病和运动障碍等神经疾病有关。此外,功能注释分析表明,一些鉴定出的蛋白质参与氧化代谢。为了进一步研究参与神经损伤的蛋白质,我们验证了两种表达下调最明显的蛋白质,即参与突触功能和神经传递的 Syn2 和 Dpysl2。我们发现 PKU 小鼠大脑中的 Glu2/3 和 NR1 受体亚基表达上调。我们的结果表明,这些蛋白质的差异表达可能与 PKU 脑功能障碍的发生过程有关,即突触可塑性降低和神经传递受损。我们鉴定了一组受高苯丙氨酸血症影响的蛋白质。我们认为,PKU 脑功能障碍也依赖于 Syn2 和 Dpysl2 水平的降低、Glu2/3 和 NR1 水平的升高以及 Pkm、Ckb、Pgam1 和 Eno1 水平的降低。这些发现最终证实,突触功能、传递和能量代谢的改变是高苯丙氨酸血症引起的脑损伤的基础。