硫酸乙酰肝素C-5-异构化酶在神经树突的形态发生中的功能及其被miRNA调控的机制研究

The defect of neuritogenesis involves in neurodegenerative diseases. Hence, study of the underlying molecular mechanisms is key step for the diagnosis and treatment of neurodegenerative diseases. Glucuronyl C5-epimerase (Glce) plays crucial role in the biosynthesis of cell membrane heparin sulfate proteoglycan. In our previous studies, we had determined the crystal structure of Glce protein. Besides, Glce implicated in NGF induced difference of PC12 cells. Overexpression or knockdown Glce affected the dendritic morphology in cortical neurons primary culture. In addition, we found that Glce neuronal conditional knockout mice had low cognitive ability. However, the functions and the regulation mechanisms of Glce are still unknown. In this project, cortical neurons primary cultures, Glce neuronal conditional knockout mice, techniques in molecular biology field will be employed to study the functions and the mechanisms of dendritic morphology, and the molecular mechanisms of Glce regulated by miRNAs. In addition, the mechanisms of enzymatic functions of Glce in dendritic morphology will also be investigated. Achievement of this project would be helpful to elucidate the mechanisms of neurodegenerative diseases and discover the key target molecule, and also provide a solid theoretical basis for targeting drug research.

神经突生成缺陷与神经退行性疾病密切相关，因此研究其发生的分子机制对疾病的诊治具有重要意义。C-5-异构化酶（Glce）是细胞膜硫酸乙酰肝素蛋白聚糖关键合成酶之一。我们早期研究确定了Glce酶的晶体结构，并发现Glce干预NGF诱导的大鼠肾上腺嗜铬细胞瘤PC12类神经细胞的分化，在鼠大脑皮层原代培养神经细胞中高表达或沉默Glce基因影响其树突的形态发生，进一步研究发现，Glce神经元条件敲除鼠显示明显认知低下。然而，其详细的功能及其调控机制尚未知晓。本项目拟运用鼠大脑皮层神经细胞和Glce条件敲除鼠结合分子生物学技术研究Glce在神经树突的形态发生中的功能机制，阐明miRNAs靶向Glce调控其相关功能的分子机制，同时探讨Glce酶学功能在神经树突的形态发生中的作用机制。该项目的实施将有助于阐明神经退行性疾病发病机理及其关键靶分子的发现，并为相关靶向性药物研究提供坚实理论基础。