Developmental neuroscienceDevelopmental regulation of nicotinic acetylcholine receptors within midbrain dopamine neurons
Section snippets
Materials
The following materials were obtained from the indicated sources: bovine serum albumin, polyvinylpyrrolidone, poly-l-lysine, RNase A, (−)nicotine bitartarate, d(+)-glucose and Hepes (Sigma-Aldrich, St. Louis, MO, USA); pBluescript II SK+ (Stratagene, La Jolla, CA, USA); anti-digoxygenin (Dig)-AP Fab antibody, Dig-UTP, Genius system nonradioactive nucleic acid detection kit, restriction enzymes, T3, T7 polymerases, proteinase K and yeast tRNA (Boehringer Mannheim Biochemicals, Indianapolis, IN,
Fetal period
Most of the subunits that are known to be expressed within the adult SNc and VTA DA neurons (Azam et al., 2002) were also present at G15, the earliest time point examined. At this early age α3, α4, α5 and β2 mRNAs were expressed at higher levels than in the adult (Fig. 1). As can be seen in the emulsion-dipped slides, which allow analysis at the neuronal level (Fig. 2), these nAChR subunits were co-expressed in TH neurons as early as G15. Furthermore, there was nAChR subunit expression in what
Discussion
The present study provides strong evidence for the presence of functional nAChRs on DA neurons even during the fetal period. As early as G15, only a couple of days following the appearance of TH within midbrain DA neurons (Specht et al 1981, Voorn et al 1988), nAChR subunit mRNAs and high affinity binding sites are present within these neurons, and functional receptors are detectable as early as G17-18. During the first three postnatal weeks, there is a gradual increase in both the efficacy and
Acknowledgments
This work was supported by NIH grant DA10612 and a Predoctoral Research Fellowship (L.A.) from the UC Tobacco Related Disease Research Program.
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