Xenopus laevis oocytes have been extensively used as a heterologous expression system for the study of ion channels. While used successfully worldwide as tool for expressing and characterizing ion channels from a wide range of species, the limited longevity of oocytes once removed from the animal can pose significant challenges. In this study, we evaluate a simple and useful method that extends the longevity of Xenopus oocytes after removal from the animal and quantitatively assessed the reliability of the electrophysiological date obtained. The receptor used for this study was the UNC-49 receptor originally isolated from the sheep parasite, Haemonchus contortus. Overall, we found that immediate storage of the ovary in supplemented ND96 storage buffer at 4 °C could extend their use for up to 17 days with almost 80% providing reliable electrophysiological data. This means that a single extraction can provide at least 3 weeks of experiments. In addition, we examined 24-day-old oocytes (week 4) extracted from a single frog and also obtained reliable data using the same approach. However, 50% of these oocytes were usable for full dose–response experiments. Overall, we did find that this method has the potential to significantly extend the use of single oocyte extractions for two-electrode voltage clamp electrophysiology.

中文翻译：

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评估手术提取的非洲爪蟾卵母细胞用于线虫配体门控离子通道的寿命。

非洲爪蟾卵母细胞已被广泛用作研究离子通道的异源表达系统。尽管在全世界范围内已成功地用作表达和表征各种物种离子通道的工具，但一旦从动物体内取出卵母细胞，其有限的寿命可能会带来巨大挑战。在这项研究中，我们评估了一种简单而有用的方法，该方法可延长从动物身上移除后爪蟾卵母细胞的寿命，并定量评估获得的电生理数据的可靠性。用于该研究的受体是最初从绵羊寄生虫Haemonchus contortus分离的UNC-49受体。。总的来说，我们发现卵巢在4°C的ND96存储缓冲液中立即存储可以延长其使用长达17天，几乎80％的卵巢提供可靠的电生理数据。这意味着一次提取可以提供至少3周的实验。此外，我们检查了从一只青蛙中提取的24天大的卵母细胞（第4周），并使用相同的方法获得了可靠的数据。但是，这些卵母细胞中有50％可用于全剂量反应实验。总的来说，我们确实发现该方法有可能极大地扩展单卵母细胞提取物在两电极电压钳电生理中的应用。