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UPA Perpustakaan Universitas Jember

Evaluating the longevity of surgically extracted Xenopus laevis oocytes for the study of nematode ligand‑gated ion channels

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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 signifcant 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 bufer 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 fnd that this method has
the potential to signifcantly extend the use of single oocyte extractions for two-electrode voltage clamp electrophysiology

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