Kulak J M, Nguyen T A, Olivera B M, McIntosh J M
Department of Biology, University of Utah, Salt Lake City, Utah, 84112, USA.
J Neurosci. 1997 Jul 15;17(14):5263-70. doi: 10.1523/JNEUROSCI.17-14-05263.1997.
Activation of presynaptic nicotinic acetylcholine receptors (nAChRs) can induce the release of neurotransmitters such as dopamine and norepinephrine in the CNS. Accumulating evidence suggests that distinct nAChR subtypes are involved; however, it has been difficult to determine the subunit composition of these receptors, in part because of the lack of a sufficient variety of selective nAChR ligands. We present experimental data that at least two different nAChR complexes are involved in dopamine release, one of which has an alpha3/beta2 subunit interface. The recently discovered peptide alpha-conotoxin MII is a potent and selective inhibitor of rat nAChRs containing an interface formed by alpha3 and beta2 subunits. We used this peptide to examine nicotine-stimulated release of dopamine from rat striatal synaptosomes and of norepinephrine from hippocampal synaptosomes. MII (100 nM) blocks 34-49% of the nicotine-stimulated dopamine release, but not dopamine release evoked by elevated [K+]. Furthermore, two peptides structurally related to alpha-conotoxin MII, namely alpha-conotoxin MI (selective for alpha1beta1gammadelta nAChRs) and alpha-conotoxin ImI (selective for alpha7-containing nAChRs), have no effect on nicotine-stimulated dopamine release. The results indicate that one third to half of the dopamine release in the striatal preparation is mediated by nAChRs with an alpha3/beta2 subunit interface. In contrast, </=10% of nicotine-stimulated release of norepinephrine from hippocampal synaptosomes is modulated by nAChRs with alpha3/beta2 subunit interfaces.
突触前烟碱型乙酰胆碱受体(nAChRs)的激活可诱导中枢神经系统中多巴胺和去甲肾上腺素等神经递质的释放。越来越多的证据表明,不同的nAChR亚型参与其中;然而,确定这些受体的亚基组成一直很困难,部分原因是缺乏足够多样的选择性nAChR配体。我们提供的实验数据表明,至少有两种不同的nAChR复合物参与多巴胺释放,其中一种具有α3/β2亚基界面。最近发现的肽α-芋螺毒素MII是大鼠nAChRs的一种强效且选择性抑制剂,该受体含有由α3和β2亚基形成的界面。我们使用这种肽来检测尼古丁刺激大鼠纹状体突触体释放多巴胺以及海马突触体释放去甲肾上腺素的情况。MII(100 nM)可阻断34% - 49%的尼古丁刺激的多巴胺释放,但不影响由升高的[K +]诱发的多巴胺释放。此外,两种与α-芋螺毒素MII结构相关的肽,即α-芋螺毒素MI(对α1β1γδ nAChRs具有选择性)和α-芋螺毒素ImI(对含α7的nAChRs具有选择性),对尼古丁刺激的多巴胺释放没有影响。结果表明,纹状体制剂中三分之一到一半的多巴胺释放是由具有α3/β2亚基界面的nAChRs介导的。相比之下,海马突触体中尼古丁刺激的去甲肾上腺素释放中,≤10%受具有α3/β2亚基界面的nAChRs调节。