Microtransplantation of ligand-gated receptor-channels from fresh or frozen nervous tissue into Xenopus oocytes: a potent tool for expanding functional information.
Despite huge improvements in neurobiological approaches for investigating the functional properties of neurotransmitter receptors and ion channels, many difficulties are still encountered when focusing on the human brain. Electrophysiological studies aimed at performing direct determinations on human nervous tissue are limited by neurosurgery and also by pathophysiological conditions prevailing before and after the resective operation. The electrophysiological study of receptors and channels becomes difficult also in animal models when the cells are not accessible and/or the experiments last many hours, during which the examined nervous tissue usually becomes unhealthy. To increase the possibility of doing optimal electrophysiological recordings, addressed to investigate the functional properties of receptors and channels, more than two decades ago, foreign mRNAs were injected into Xenopus oocytes to heterologously express the receptors; and about a decade ago cell membranes were injected into the oocytes to directly transplant the native receptors. While the first approach needs complex procedures for mRNA isolation, the membrane preparations are simpler to obtain and the embedded receptors are transplanted in their own membrane, with their own glycosylation and together with any ancillary proteins they may have. Using injections of membranes isolated from fresh nervous tissues several issues have already been addressed and many questions can be answered in the near future. Strikingly, with this approach it has been possible to "resuscitate" receptors and ion channels from tissues kept frozen for many years. This review focuses on recently obtained information and on some new lines of biological research using receptor microtransplantation into oocytes.
Eusebi
F
F
Dipartimento di Fisiologia e Farmacologia-Centro di Eccellenza BEMM, Universita' di Roma Sapienza, Piazzale A. Moro 5, I00185 Roma, Italy. fabrizio.eusebi@uniroma1.it
Palma
E
E
Amici
M
M
Miledi
R
R
D000818:Animals; D001921:Brain / Q000502:physiology*; D016380:Brain Tissue Transplantation / Q000379:methods*; D005260:Female; D005615:Freezing; D006801:Humans; D015640:Ion Channel Gating / Q000502:physiology; D007473:Ion Channels / Q000378:metabolism*; D009865:Oocytes / Q000502:physiology*; D014982:Xenopus laevis / Q000502:physiology*
D007473:Ion Channels