Cristofori-Armstrong Ben, Soh Ming S, Talwar Sahil, Brown Darren L, Griffin John D O, Dekan Zoltan, Stow Jennifer L, King Glenn F, Lynch Joseph W, Rash Lachlan D
Institute for Molecular Bioscience, The University of Queensland, St Lucia, QLD 4072, Australia.
Queensland Brain Institute, The University of Queensland, St Lucia, QLD 4072, Australia.
Sci Rep. 2015 Oct 6;5:14763. doi: 10.1038/srep14763.
For the past 30 years, oocytes from Xenopus laevis have been extensively used to express and characterise ion channels in an easily controlled environment. Here we report the first use of oocytes from the closely related species Xenopus borealis as an alternative expression system for neuronal ion channels. Using the two-electrode voltage-clamp technique, we show that a wide variety of voltage- and ligand-gated ion channels have the same channel properties and pharmacological profiles when expressed in either X. laevis or X. borealis oocytes. Potential advantages of the X. borealis oocytes include a smaller endogenous chloride current and the ability to produce more intense fluorescence signals when studied with voltage-clamp fluorometry. Scanning electron microscopy revealed a difference in vitelline membrane structure between the two species, which may be related to the discrepancy in fluorescence signals observed. We demonstrate that X. borealis oocytes are a viable heterologous system for expression of neuronal ion channels with some potential advantages over X. laevis oocytes for certain applications.
在过去30年里，非洲爪蟾的卵母细胞被广泛用于在易于控制的环境中表达和表征离子通道。在此，我们报告首次使用亲缘关系相近的北方爪蟾的卵母细胞作为神经元离子通道的替代表达系统。使用双电极电压钳技术，我们发现，当在非洲爪蟾或北方爪蟾的卵母细胞中表达时，多种电压门控离子通道和配体门控离子通道具有相同的通道特性和药理学特征。北方爪蟾卵母细胞的潜在优势包括较小的内源性氯离子电流，以及在用电压钳荧光测定法研究时产生更强荧光信号的能力。扫描电子显微镜显示这两个物种的卵黄膜结构存在差异，这可能与观察到的荧光信号差异有关。我们证明，北方爪蟾卵母细胞是用于表达神经元离子通道的可行异源系统，在某些应用中比非洲爪蟾卵母细胞具有一些潜在优势。
