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

Sterling N. Sudweeks and Jerrel L. Yakel

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 2-7 and 2-4, 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 4, 5, 7 and 2-4 in stratum radiatum interneurons; 2, 3, 4, 7, 2 and 3 subunits in stratum oriens interneurons; and 2-4 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 3 + 5, 4 + 2, 4 + 3, 7 + 2, 2 + 3 and 3 + 4.

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 3, 7 and 2 nAChR subunits were individually correlated with a fast rise time, the 2 nAChR subunit was correlated with a medium rise time, and the 4 nAChR subunit was correlated with a slow rise time. The 2 and 4 nAChR subunits were also significantly correlated with slow desensitization kinetics.

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