Functional Expression Of Nicotinic Acetylcholine Receptors Containing Rat Î±7 Subunits In Human Sh-Sy5Y Neuroblastoma Cells
Neuronal nicotinic acetylcholine receptors (nAChR) are made from different combinations of subunits encoded by a diverse family of genes. However, the recently cloned Î±7 gene codes for subunits that can form homooligomeric nAChR complexes when expressed in Xenopus oocytes. Electrophysiological studies reveal that these Î±7-nAChR function as Î±-bungarotoxin (Bgt)-sensitive, quickly activating/inactivating ion channels with a unique pharmacological profile and an unusually high permeability to calcium ions. Although similar observations have been made in studies of Bgt-sensitive, functional nAChR subtypes that are naturally expressed in neuronal cells, all attempts until now to reconstitute functional Î±7-nAChR in cell lines have failed. Here we report the successful use of SH-SY5Y human neuroblastoma cells, which naturally express low levels of endogenous Î±7 transcripts, to stably overexpress heterologous rat nAChR Î±7 transgenes. These transgenes are expressed as the appropriately-sized Î±7 messages and protein, and stably transfected SH-SY5Y cells have over 30-times higher levels of specific Bgt binding sites than do wild-type cells. Whole cell current recordings confirm that transfected cells express functional nAChR that are sensitive to blockade by Bgt and display the typical physiological and pharmacological profiles of Î±7-nAChR. We conclude that stable, functional expression of Î±7 transgenes in a mammalian cell line has been achieved for the first time. Â© 1994.
Digital Object Identifier (DOI)
Puchacz, Elzbieta; Buisson, Bruno; Bertrand, Daniel; and Lukas, Ronald J., "Functional Expression Of Nicotinic Acetylcholine Receptors Containing Rat Î±7 Subunits In Human Sh-Sy5Y Neuroblastoma Cells" (1994). Neurobiology. 222.