Plasmid Vector Expression of Alpha6 nAChR Isoforms in Xenopus Oocytes
Abstract
Nicotinic acetylcholine receptors (nAChRs) containing alpha6 including alpha6 beta2 beta3 isoforms are important for dopamine transmission. Receptors containing alpha6, especially the alpha6 beta3 beta2 isoform, are hypothesized to be a contender for nigrostriatal pathway (NSP) dysfunction in Parkinson’s patients. NSP dysfunction is responsible for problems receiving and turning nAChR messages into dopamine transmission resulting in dopamine deficiency and hypokinesia. Presently, the plasmid vectors we are using offer a limited expression ability for receptors containing alpha6 in Xenopus oocytes. This problem has motivated us to find plasmid vectors which yield high expression of alpha6 isoforms. Successful subcloning of alpha6 into high expression plasmid vectors would allow expression of alpha6 isoforms including alpha6 beta3 beta2 in Xenopus oocytes. Functional voltage clamp testing will be done to provide greater understanding of isoforms responsible for NSP dysfunction. This information will facilitate testing of computational model accuracy which would verify pharmacological treatment to hopefully surpass the efficacy of current treatment for Parkinson’s.
Plasmid Vector Expression of Alpha6 nAChR Isoforms in Xenopus Oocytes
Nicotinic acetylcholine receptors (nAChRs) containing alpha6 including alpha6 beta2 beta3 isoforms are important for dopamine transmission. Receptors containing alpha6, especially the alpha6 beta3 beta2 isoform, are hypothesized to be a contender for nigrostriatal pathway (NSP) dysfunction in Parkinson’s patients. NSP dysfunction is responsible for problems receiving and turning nAChR messages into dopamine transmission resulting in dopamine deficiency and hypokinesia. Presently, the plasmid vectors we are using offer a limited expression ability for receptors containing alpha6 in Xenopus oocytes. This problem has motivated us to find plasmid vectors which yield high expression of alpha6 isoforms. Successful subcloning of alpha6 into high expression plasmid vectors would allow expression of alpha6 isoforms including alpha6 beta3 beta2 in Xenopus oocytes. Functional voltage clamp testing will be done to provide greater understanding of isoforms responsible for NSP dysfunction. This information will facilitate testing of computational model accuracy which would verify pharmacological treatment to hopefully surpass the efficacy of current treatment for Parkinson’s.