Bianchi Laura, Driscoll Monica
Department of Molecular Biology and Biochemistry, Rutgers University, Piscataway, NJ 08854, USA.
WormBook. 2006 Aug 1:1-16. doi: 10.1895/wormbook.1.117.1.
Physiological methods entered the world of C. elegans, a model system used for many years to study development and a plethora of biological processes mainly employing genetic, molecular and anatomical techniques. One of the methods introduced by physiologists is the use of Xenopus oocytes for expression of C. elegans ion channels. Oocytes of the South African frog Xenopus laevis are used widely for the expression of mammalian channels and transporters contributing to numerous discoveries in these fields. They now promise to aid C. elegans researchers in deciphering mechanisms of channels function and regulation with implications for mammalian patho-physiology. Heterologous cRNA can be easily injected into Xenopus oocytes and translated proteins can be studied using several techniques including electrophysiology, immunocytochemistry and protein biochemistry. This chapter will focus on techniques used for oocyte preparation and injection, and will give a brief overview of specific methods. Limitations of the use of Xenopus oocytes will be also discussed.
生理学方法进入了秀丽隐杆线虫的研究领域,秀丽隐杆线虫是一个多年来用于研究发育以及众多主要采用遗传学、分子学和解剖学技术的生物学过程的模式系统。生理学家引入的方法之一是利用非洲爪蟾卵母细胞来表达秀丽隐杆线虫的离子通道。南非爪蟾(非洲爪蟾)的卵母细胞被广泛用于哺乳动物通道和转运体的表达,这为这些领域的众多发现做出了贡献。它们现在有望帮助秀丽隐杆线虫研究人员解读通道功能和调节机制,这对哺乳动物病理生理学具有重要意义。异源cRNA可以很容易地注射到非洲爪蟾卵母细胞中,并且可以使用包括电生理学、免疫细胞化学和蛋白质生物化学在内的多种技术来研究翻译后的蛋白质。本章将重点介绍用于卵母细胞制备和注射的技术,并简要概述具体方法。还将讨论使用非洲爪蟾卵母细胞的局限性。