Radley Jason J, Farb Claudia R, He Yong, Janssen William G M, Rodrigues Sarina M, Johnson Luke R, Hof Patrick R, LeDoux Joseph E, Morrison John H
Department of Neuroscience, Mount Sinai School of Medicine, New York, NY 10029, USA.
Brain Res. 2007 Feb 23;1134(1):87-94. doi: 10.1016/j.brainres.2006.11.045. Epub 2007 Jan 17.
Synapses onto dendritic spines in the lateral amygdala formed by afferents from the auditory thalamus represent a site of plasticity in Pavlovian fear conditioning. Previous work has demonstrated that thalamic afferents synapse onto LA spines expressing glutamate receptor (GluR) subunits, but the GluR subunit distribution at the synapse and within the cytoplasm has not been characterized. Therefore, we performed a quantitative analysis for alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) receptor subunits GluR2 and GluR3 and N-methyl-D-aspartate (NMDA) receptor subunits NR1 and NR2B by combining anterograde labeling of thalamo-amygdaloid afferents with postembedding immunoelectron microscopy for the GluRs in adult rats. A high percentage of thalamo-amygdaloid spines was immunoreactive for GluR2 (80%), GluR3 (83%), and NR1 (83%), while a smaller proportion of spines expressed NR2B (59%). To compare across the various subunits, the cytoplasmic to synaptic ratios of GluRs were measured within thalamo-amygdaloid spines. Analyses revealed that the cytoplasmic pool of GluR2 receptors was twice as large compared to the GluR3, NR1, and NR2B subunits. Our data also show that in the adult brain, the NR2B subunit is expressed in the majority of in thalamo-amygdaloid spines and that within these spines, the various GluRs are differentially distributed between synaptic and non-synaptic sites. The prevalence of the NR2B subunit in thalamo-amygdaloid spines provides morphological evidence supporting its role in the fear conditioning circuit while the differential distribution of the GluR subtypes may reflect distinct roles for their involvement in this circuitry and synaptic plasticity.
来自听觉丘脑的传入神经在外侧杏仁核的树突棘上形成的突触代表了经典条件恐惧反射中的一个可塑性位点。先前的研究表明,丘脑传入神经与表达谷氨酸受体(GluR)亚基的外侧杏仁核树突棘形成突触,但尚未对突触处和细胞质内的GluR亚基分布进行表征。因此,我们通过将丘脑 - 杏仁核传入神经的顺行标记与成年大鼠GluR的包埋后免疫电子显微镜相结合,对α-氨基-3-羟基-5-甲基-4-异恶唑丙酸(AMPA)受体亚基GluR2和GluR3以及N-甲基-D-天冬氨酸(NMDA)受体亚基NR1和NR2B进行了定量分析。高比例的丘脑 - 杏仁核树突棘对GluR2(80%)、GluR3(83%)和NR1(83%)呈免疫反应性,而表达NR2B的树突棘比例较小(59%)。为了比较不同亚基,我们测量了丘脑 - 杏仁核树突棘内GluR的细胞质与突触比率。分析表明,GluR2受体的细胞质池比GluR3、NR1和NR2B亚基大两倍。我们的数据还表明,在成人大脑中,NR2B亚基在大多数丘脑 - 杏仁核树突棘中表达,并且在这些树突棘内,各种GluR在突触和非突触部位之间存在差异分布。NR2B亚基在丘脑 - 杏仁核树突棘中的普遍存在提供了形态学证据,支持其在恐惧条件反射回路中的作用,而GluR亚型的差异分布可能反映了它们在该回路和突触可塑性中的不同作用。