Ivorra Isabel, Alberola-Die Armando, Cobo Raúl, González-Ros José Manuel, Morales Andrés
Departamento de Fisiología, Genética y Microbiología, Universidad de Alicante, Apdo 99, E-03080 Alicante, Spain.
Instituto de Investigación, Desarrollo e Innovación en Biotecnología Sanitaria de Elche (IDiBE), Universidad Miguel Hernández, E-03202 Elche, Spain.
Membranes (Basel). 2022 Oct 11;12(10):986. doi: 10.3390/membranes12100986.
The use of oocytes in electrophysiological and biophysical research constitutes a long and successful story, providing major advances to the knowledge of the function and modulation of membrane proteins, mostly receptors, ion channels, and transporters. Earlier reports showed that these cells are capable of correctly expressing heterologous proteins after injecting the corresponding mRNA or cDNA. More recently, the oocyte has become an outstanding host-cell model to carry out detailed studies on the function of fully-processed foreign membrane proteins after their microtransplantation to the oocyte. This review focused on the latter overall process of transplanting foreign membrane proteins to the oocyte after injecting plasma membranes or purified and reconstituted proteins. This experimental approach allows for the study of both the function of mature proteins, with their native stoichiometry and post-translational modifications, and their putative modulation by surrounding lipids, mostly when the protein is purified and reconstituted in lipid matrices of defined composition. Remarkably, this methodology enables functional microtransplantation to the oocyte of membrane receptors, ion channels, and transporters from different sources including human post-mortem tissue banks. Despite the large progress achieved over the last decades on the structure, function, and modulation of neuroreceptors and ion channels in healthy and pathological tissues, many unanswered questions remain and, most likely, oocytes will continue to help provide valuable responses.
卵母细胞在电生理和生物物理研究中的应用由来已久且成果丰硕，为膜蛋白（主要是受体、离子通道和转运蛋白）的功能及调节机制研究带来了重大进展。早期报告表明，在注射相应的mRNA或cDNA后，这些细胞能够正确表达异源蛋白。最近，卵母细胞已成为一种出色的宿主细胞模型，用于在将完全加工的外源膜蛋白微移植到卵母细胞后，对其功能进行详细研究。本综述聚焦于在注射质膜或纯化及重组蛋白后，将外源膜蛋白移植到卵母细胞的后一整体过程。这种实验方法既可以研究具有天然化学计量和翻译后修饰的成熟蛋白的功能，也可以研究其可能受到周围脂质的调节，尤其是当蛋白在特定组成的脂质基质中纯化和重组时。值得注意的是，这种方法能够将来自不同来源（包括人类尸检组织库）的膜受体、离子通道和转运蛋白功能微移植到卵母细胞中。尽管在过去几十年里，健康和病理组织中神经受体和离子通道的结构、功能及调节方面取得了很大进展，但仍有许多问题未得到解答，而且很可能卵母细胞将继续有助于提供有价值的答案。
