Barenco Maria Grazia, Valori Chiara F, Roncoroni Chiara, Loewer Johannes, Montrasio Fabio, Rossi Daniela
Prion Research Group, Paul-Ehrlich-Institut, Langen, Germany.
J Neurosci Res. 2009 Feb 15;87(3):806-19. doi: 10.1002/jnr.21894.
The cellular prion protein (PrP(C)) is a highly conserved glycoprotein of unknown biological function. To gain insight into the physiological role of PrP(C), we generated a novel PrP knockout cell line, named PrP(o/o) ML, by immortalization of neuroepithelial precursor cells derived from the cerebellum of PrP-knockout mice using the temperature-sensitive simian virus 40 (SV40) large T antigen. We demonstrated that the PrP(o/o) ML cell line is a unipotent precursor line with glutamatergic properties, which can acquire neuronal features when cultivated under specific conditions. The role of the prion protein in the process of neuronal differentiation was then analyzed in the PrP(o/o) ML cells reconstituted with either the full-length or an amino-terminally deleted form of the prion protein. We show that the expression of PrP(C) facilitates the processes of neuronal differentiation and neuritogenesis and that the deletion of its amino-terminal domain reduces the efficiency, but does not suppress this activity. This cell line represents a useful tool for studying PrP-dependent signal transduction pathways during differentiation of neuronal stem/precursor cells.
细胞朊蛋白（PrP(C)）是一种生物学功能未知的高度保守糖蛋白。为深入了解PrP(C)的生理作用，我们通过使用温度敏感型猿猴病毒40（SV40）大T抗原永生化源自PrP基因敲除小鼠小脑的神经上皮前体细胞，构建了一种新型的PrP基因敲除细胞系，命名为PrP(o/o) ML。我们证明PrP(o/o) ML细胞系是具有谷氨酸能特性的单能前体细胞系，在特定条件下培养时可获得神经元特征。然后，在分别用全长或氨基末端缺失形式的朊蛋白重构的PrP(o/o) ML细胞中分析朊蛋白在神经元分化过程中的作用。我们发现PrP(C)的表达促进神经元分化和神经突形成过程，其氨基末端结构域的缺失会降低效率，但不会抑制这种活性。该细胞系是研究神经元干/前体细胞分化过程中PrP依赖信号转导途径的有用工具。
