Elsevier

Neuroscience

Volume 144, Issue 4, 23 February 2007, Pages 1347-1360
Neuroscience

Developmental neuroscience
Developmental regulation of nicotinic acetylcholine receptors within midbrain dopamine neurons

https://doi.org/10.1016/j.neuroscience.2006.11.011Get rights and content

Abstract

We have combined anatomical and functional methodologies to provide a comprehensive analysis of the properties of nicotinic acetylcholine receptors (nAChRs) on developing dopamine (DA) neurons of Sprague–Dawley rats. Double-labeling in situ hybridization was used to examine the expression of nAChR subunit mRNAs within developing midbrain DA neurons. As brain maturation progressed there was a change in the pattern of subunit mRNA expression within DA neurons, such that α3 and α4 subunits declined and α6 mRNA increased. Although there were strong similarities in subunit mRNA expression in substantia nigra (SNc) and ventral tegmental area (VTA), there was higher expression of α4 mRNA in SNc than VTA at gestational day (G) 15, and of α5, α6 and β3 mRNAs during postnatal development. Using a superfusion neurotransmitter release paradigm to functionally characterize nicotine-stimulated release of [3H]DA from striatal slices, the properties of the nAChRs on DA terminals were also found to change with age. Functional nAChRs were detected on striatal terminals at G18. There was a decrease in maximal release in the first postnatal week, followed by an increase in nicotine efficacy and potency during the second and third postnatal weeks. In the transition from adolescence (postnatal days (P) 30 and 40) to adulthood, there was a complex pattern of functional maturation of nAChRs in ventral, but not dorsal, striatum. In males, but not females, there were significant changes in both nicotine potency and efficacy during this developmental period. These findings suggest that nAChRs may play critical functional roles throughout DA neuronal maturation.

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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