Department of Anatomy, Howard University College of Medicine, Washington, DC 20059, USA.
Neuroscience. 2011 Aug 11;188:168-81. doi: 10.1016/j.neuroscience.2011.04.069. Epub 2011 May 12.
Untimely activation of nicotinic acetylcholine receptors (nAChRs) by nicotine results in short- and long-term consequences on learning and behavior. In this study, the aim was to determine how prenatal nicotine exposure affects components of glutamatergic signaling in the hippocampus during postnatal development. We investigated regulation of both nAChRs and glutamate receptors for AMPA and N-methyl-D-aspartate (NMDA), from postnatal day 1 (P1) to P63 after a temporally restricted exposure to saline or nicotine for 14 days in utero. We analyzed postsynaptic density components associated with AMPA receptor (AMPAR) and NMDA receptor (NMDAR) signaling: calmodulin (CaM), CaM Kinase II alpha (CaMKIIα), and postsynaptic density-95 (PSD95), as well as presynaptically localized synaptosomal-associated protein 25 (SNAP25). At P1, there was significantly heightened expression of AMPAR subunit GluR1 but not GluR2, and of NMDAR subunits NR1, NR2a, and NR2d but not NR2b. NR2c was not detectable. CaM, CaMKIIα, and PSD95 were also significantly upregulated at P1, together with presynaptic SNAP25. This enhanced expression of glutamate receptors and signaling proteins was concomitant with elevated levels of [³H]epibatidine (³H]EB) binding in prenatal nicotine-exposed hippocampus, indicating that α4β2 nAChR may influence glutamatergic function in the hippocampus at P1. By P14, neither [³H]EB binding nor the expression levels of subunits GluR1, GluR2, NR1, NR2a, NR2b, NR2c, or NR2d seemed changed with prenatal nicotine. However, CaMKIIα was significantly upregulated with nicotine treatment while CaM showed downregulation at P14. The effects of nicotine persisted in P63 young adult brains which exhibited significantly downregulated GluR2, NR1, and NR2c expression levels in hippocampal homogenates and a considerably muted overall distribution of [³H]AMPA binding in areas CA1, CA2 and CA3, and the dentate gyrus. Our results suggest that prenatal nicotine exposure can regulate the glutamatergic signaling system throughout postnatal development by enhancing or inhibiting availability of AMPAR and NMDAR or their signaling components. The persistent depression, in adults, of the requisite NR1 subunit for NMDAR assembly, and of GluR2, important for assembly, trafficking, and biophysical properties of AMPAR, indicates that nicotine may alter ionotropic glutamate receptor stoichiometry and functional properties in adults after prenatally restricted nicotine exposure.
尼古丁会过早激活烟碱型乙酰胆碱受体(nAChRs),从而对学习和行为产生短期和长期影响。在这项研究中,我们的目的是确定宫内暴露于尼古丁对出生后发育过程中海马体内谷氨酸能信号传导的组成部分的影响。我们从产后第 1 天(P1)至第 63 天(P63),通过在宫内 14 天进行时间限制的盐水或尼古丁暴露,调查了 nAChR 和谷氨酸受体(AMPA 和 N-甲基-D-天冬氨酸(NMDA))的调节。我们分析了与 AMPA 受体(AMPAR)和 NMDA 受体(NMDAR)信号相关的突触后密度成分:钙调蛋白(CaM)、钙调蛋白激酶 IIα(CaMKIIα)和突触后密度蛋白 95(PSD95),以及突触前定位的突触相关蛋白 25(SNAP25)。在 P1 时,AMPAR 亚基 GluR1 但不是 GluR2,以及 NMDAR 亚基 NR1、NR2a、NR2d 但不是 NR2b 的表达明显升高。NR2c 不可检测。CaM、CaMKIIα 和 PSD95 也在 P1 时显著上调,与突触前 SNAP25 一起上调。这种谷氨酸受体和信号蛋白的增强表达伴随着[³H]epibatidine([³H]EB)结合物在宫内尼古丁暴露的海马体中的水平升高,表明α4β2 nAChR 可能在 P1 时影响海马体中的谷氨酸能功能。到 P14 时,[³H]EB 结合物的水平以及亚基 GluR1、GluR2、NR1、NR2a、NR2b、NR2c 或 NR2d 的表达水平似乎都没有因宫内尼古丁而改变。然而,CaMKIIα 在尼古丁处理时明显上调,而 CaM 在 P14 时下调。尼古丁的影响在 P63 年轻成年大脑中持续存在,其海马体匀浆中的 GluR2、NR1 和 NR2c 表达水平显著下调,并且 [³H]AMPA 结合的整体分布在 CA1、CA2 和 CA3 区域以及齿状回区域明显减弱。我们的结果表明,宫内尼古丁暴露可以通过增强或抑制 AMPAR 和 NMDAR 或其信号成分的可用性来调节出生后发育过程中的谷氨酸能信号系统。成年后 NMDA 组装所必需的 NR1 亚基和 GluR2 的持续下调,GluR2 对于 AMPAR 的组装、运输和生物物理特性很重要,这表明尼古丁可能会改变出生后限制尼古丁暴露后成年离子型谷氨酸受体的化学计量和功能特性。