MRNA-directed synthesis and insertion of functional amino acid receptors in Xenopus laevis oocytes.

Abstract

A major goal for neuroscientists over the past 1&20 years has been the understanding of the mechanism and control of membrane excitability in the mammalian central nervous system (CNS). To realize this ambition, various methods and techniques have been evolved to study drugand voltage-operated membrane ion channels, which have required the manipulation of CNS tissue. A large majority of these techniques, both biochemical and electrophysiological, have involved the use of brain slices, membrane homogenates and also dissociated and organotypic tissue culture preparations. In all three situations, the drug receptors and associated ionophores resided in the native neuronal/glial membrane and were not always ideally placed for study. As an alternative, we have effectively transferred some of the apparatus mediating CNS membrane excitability, such as certain drug receptors and voltage-activated ion channels, from their neuronal environment, into the cell membrane of the Xenopus oocyte, by utilizing the translation of exogenous mRNA. This preparation has been a highly convenient medium for electrophysiological and radioligand-binding studies on drug