Microtransplantation of neurotransmitter receptors from cells to Xenopus oocyte membranes: new procedure for ion channel studies.
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.
Miledi
Ricardo
R
Instituto de Neurobiología, Campus UNAM-Juriquilla, México.
Palma
Eleonora
E
Eusebi
Fabrizio
F
D000818:Animals; D016380:Brain Tissue Transplantation / Q000379:methods / Q000502:physiology*; D002460:Cell Line; D002462:Cell Membrane / Q000502:physiology / Q000637:transplantation*; D006801:Humans; D015640:Ion Channel Gating / Q000502:physiology*; D009865:Oocytes / Q000502:physiology*; D017981:Receptors, Neurotransmitter / Q000502:physiology*; D014982:Xenopus laevis
D017981:Receptors, Neurotransmitter