Cailliau Katia, Browaeys-Poly Edith
Université des Sciences et Technologies de Lille, Laboratoire de Régulation des Signaux de Division, Villeneuve d'Ascq Cedex, France.
Methods Mol Biol. 2009;518:43-55. doi: 10.1007/978-1-59745-202-1_4.
To study protein function in cellular signaling, manual microinjection is a direct technique, but limited by the small size of many cells. The giant vertebrate cell, the Xenopus laevis oocyte, is a perfect model system to perform these studies. Oocytes are numerous and synchronous cells, arrested in the G2 phase of the cell cycle and easily amenable to biochemical, electrophysiological, and cytological studies. We describe how to microinject proteins or peptides in this model and we study, as an example, the Grb2 transduction cascade.
为了研究细胞信号传导中的蛋白质功能,手动显微注射是一种直接的技术,但受到许多细胞体积小的限制。大型脊椎动物细胞——非洲爪蟾卵母细胞,是进行这些研究的理想模型系统。卵母细胞数量众多且同步,停滞在细胞周期的G2期,易于进行生化、电生理和细胞学研究。我们描述了如何在这个模型中显微注射蛋白质或肽,并以Grb2转导级联为例进行研究。