Miledi Ricardo, Palma Eleonora, Eusebi Fabrizio
Instituto de Neurobiología, Campus UNAM-Juriquilla, México.
Methods Mol Biol. 2006;322:347-55. doi: 10.1007/978-1-59745-000-3_24.
The Xenopus oocyte is largely used as a cell expression system for studying both structure and function of transmitter receptors and ion channels. Messenger RNA extracted from the brain and injected into oocytes leads to the synthesis and membrane incorporation of many types of functional ion channels. A new method was developed further to transplant neurotransmitter receptors from human brain or cultured cell lines to the membrane of Xenopus oocytes. This method represents a modification of the method used many years ago of injecting into oocytes membrane vesicles from Torpedo electroplaques, yielding the expression of functional Torpedo acetylcholine receptors. We describe this approach by extracting membrane vesicles from human hippocampus or temporal neocortex and from mammalian cell lines stably expressing glutamate or neuronal nicotinic receptors. Because the human neurotransmitter receptors are "microtransplanted" with their native cell membranes, this method extends the usefulness of Xenopus oocytes as an expression system for addressing issues in many fields, including channelopathies.
非洲爪蟾卵母细胞在很大程度上被用作一种细胞表达系统，用于研究递质受体和离子通道的结构与功能。从大脑中提取并注射到卵母细胞中的信使核糖核酸会导致多种功能性离子通道的合成与膜整合。进一步开发了一种新方法，将来自人类大脑或培养细胞系的神经递质受体移植到非洲爪蟾卵母细胞的膜上。这种方法是对多年前使用的将电鳐电板的膜囊泡注射到卵母细胞中的方法的改进，从而产生功能性电鳐乙酰胆碱受体的表达。我们通过从人类海马体或颞叶新皮层以及稳定表达谷氨酸或神经元烟碱受体的哺乳动物细胞系中提取膜囊泡来描述这种方法。由于人类神经递质受体与其天然细胞膜一起被“微移植”，这种方法扩展了非洲爪蟾卵母细胞作为一种表达系统在解决包括通道病在内的许多领域问题方面的用途。
