Microtransplantation of membranes from cultured cells to Xenopus oocytes: a method to study neurotransmitter receptors embedded in native lipids.
The Xenopus oocyte is used as a convenient cell expression system to study the structure and function of heterogenic transmitter receptors and ion channels. Recently, we introduced a method to microtransplant already assembled neurotransmitter receptors from the human brain to the plasma membrane of Xenopus oocytes. The same approach was used here to transplant neurotransmitter receptors expressed from cultured cells to the oocytes. Membrane vesicles prepared from a human embryonic kidney cell line (HEK293) stably expressing the rat glutamate receptor 1 were injected into oocytes, and, within a few hours, the oocyte plasma membrane acquired alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid-type glutamate receptors, which had the same properties as those expressed in the original HEK cells. Analogously, oocytes injected with membranes prepared from rat pituitary GH(4)C1 cells, stably expressing homomeric human neuronal alpha 7 nicotinic acetylcholine receptors (alpha 7-AcChoRs), incorporated in their plasma membrane AcChoRs that behaved as those expressed in GH(4)C1 cells. Similar results were obtained with HEK cells stably expressing heteromeric human neuronal alpha 4 beta 2-AcChoRs. All this makes the Xenopus oocyte a powerful tool for detailed investigations of receptors and other proteins expressed in the membrane of cultured cells.
Palma
Eleonora
E
Istituto Pasteur-Fondazione Cenci Bolognetti and Dipartimento di Fisiologia Umana e Farmacologia, Universita' di Roma La Sapienza, Piazzale Aldo Moro 5, I-00185 Rome, Italy.
Trettel
Flavia
F
Fucile
Sergio
S
Renzi
Massimiliano
M
Miledi
Ricardo
R
Eusebi
Fabrizio
F
D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D017690:Cell Transplantation; D004305:Dose-Response Relationship, Drug; D006801:Humans; D007477:Ions; D050356:Lipid Metabolism; D018377:Neurotransmitter Agents / Q000378:metabolism; D009865:Oocytes / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D011950:Receptors, Cholinergic / Q000378:metabolism; D017470:Receptors, Glutamate / Q000378:metabolism; D013997:Time Factors; D014982:Xenopus laevis
D007477:Ions; D018377:Neurotransmitter Agents; D011950:Receptors, Cholinergic; D017470:Receptors, Glutamate