Functional and molecular characterization of neuronal nicotinic ACh receptors in rat CA1 hippocampal neurons

Abstract

1 The molecular and functional properties of neuronal nicotinic acetylcholine receptors (nAChRs) were characterized from CA1 neurons in rat hippocampal slices using single-cell reverse-transcription polymerase chain reaction (RT-PCR) in conjunction with whole-cell patch-clamp recordings. 2 We analysed the presence of the neuronal nAChR subunit mRNAs α2–7 and β2–4, along with the mRNA for the GABAergic markers GAD (glutamic acid decarboxylase) 65 and 67 isoforms, and VGAT (vesicular GABA transporter) in interneurons from the stratum radiatum and stratum oriens, and in CA1 pyramidal neurons. Functional nAChR-mediated currents were detected in both interneuron populations, but not in pyramidal neurons. 3 The neuronal nAChR subunit mRNAs detected in > 20 % of the populations examined were α4, α5, α7 and β2–4 in stratum radiatum interneurons; α2, α3, α4, α7, β2 and β3 subunits in stratum oriens interneurons; and β2–4 in pyramidal neurons. High levels of GABAergic marker mRNAs were detected in both interneuron populations, but not in pyramidal neurons. 4 Significant co-expression of nAChR subunits within individual neurons was detected for α3 +α5, α4 +β2, α4 +β3, α7 +β2, β2 +β3 and β3 +β4. 5 The kinetics of the nAChR-mediated currents in response to the application of 100 μm ACh were significantly correlated with the expression of particular nAChR subunits. The α3, α7 and β2 nAChR subunits were individually correlated with a fast rise time, the α2 nAChR subunit was correlated with a medium rise time, and the α4 nAChR subunit was correlated with a slow rise time. The α2 and α4 nAChR subunits were also significantly correlated with slow desensitization kinetics.