Expression and Function of Alpha3 and Beta2 Neuronal Nicotinic Acetylcholine Receptor Subunits in HEK-293 Cells

Single-cell real-time quantitative RT-PCR was used to characterize the mRNA expression of rat neuronal nicotinic acetylcholine receptor (nAChR) subunits α3 and β2 in CA1 hippocampus stratum radiatum and stratum oriens interneurons. α3β2 co-expression was detected in 43% of interneurons analyzed. The nAChR subtype α3β2 was transiently expressed in cells derived from the human embryonic kidney cell line 293 at mRNA levels found in the CA1. The functional properties of α3β2 in HEK-293 cells were characterized by whole-cell patch clamping using acetylcholine (ACh) as an agonist. The kinetics of α3β2 channels were further analyzed by altering the level of α3 DNA transfected into HEK-293 cells. Varying the α3 concentration by more than 100,000 fold did not significantly alter the majority of the kinetics; the 10%-90% rise-time was the main characteristic found to be significantly different. A decrease in α3 concentration illustrated a significant increase in rise time. This and future studies will further our understanding of the extensive role neuronal nAChRs play in modulating hippocampal activity and consequently influencing cognition and memory.

Name thesis:degree_name

MS

Grantor dc:publisher

Brigham Young University - Provo

Author dc:creator

• Steinhafel, Nathan W.

• hippocampus
• CA1
• interneurons
• nicotinic acetylcholine receptor
• HEK-293
• PCR
• patch clamp
• Cell and Developmental Biology
• Physiology

Language dc:language

English

Repository record dc:identifier

https://scholarsarchive.byu.edu/etd/1114

OAI identifier oai:identifier

oai:scholarsarchive.byu.edu:etd-2113