Burger R Michael, Pfeiffer Joshua D, Westrum Lesnick E, Bernard Amy, Rubel Edwin W
Virginia Merrill Bloedel Hearing Research Center and Department of Otolaryngology-Head and Neck Surgery, University of Washington, Seattle, Washington 98195, USA.
J Comp Neurol. 2005 Aug 15;489(1):11-22. doi: 10.1002/cne.20607.
Nucleus magnocellularis (NM), nucleus angularis (NA), and nucleus laminaris (NL), second- and third-order auditory neurons in the avian brainstem, receive GABAergic input primarily from the superior olivary nucleus (SON). Previous studies have demonstrated that both GABA(A) and GABA(B) receptors (GABA(B)Rs) influence physiological properties of NM neurons. We characterized the distribution of GABA(B)R expression in these nuclei during development and after deafferentation of the excitatory auditory nerve (nVIII) inputs. We used a polyclonal antibody raised against rat GABA(B)Rs in the auditory brainstem during developmental periods that are thought to precede and include synaptogenesis of GABAergic inputs. As early as embryonic day (E)14, dense labeling is observed in NA, NM, NL, and SON. At earlier ages immunoreactivity is present in somas as diffuse staining with few puncta. By E21, when the structure and function of the auditory nuclei are known to be mature, GABA(B) immunoreactivity is characterized by dense punctate labeling in NM, NL, and a subset of NA neurons, but label is sparse in the SON. Removal of the cochlea and nVIII neurons in posthatch chicks resulted in only a small decrease in immunoreactivity after survival times of 14 or 28 days, suggesting that a major proportion of GABA(B)Rs may be expressed postsynaptically or on GABAergic terminals. We confirmed this interpretation with immunogold TEM, where expression at postsynaptic membrane sites is clearly observed. The characterization of GABA(B)R distribution enriches our understanding of the full complement of inhibitory influences on central auditory processing in this well-studied neuronal circuit.
巨细胞核(NM)、角状核(NA)和层状核(NL)是禽脑桥中的二级和三级听觉神经元,主要从橄榄上核(SON)接收GABA能输入。先前的研究表明,GABA(A)和GABA(B)受体(GABA(B)Rs)均影响NM神经元的生理特性。我们对这些核团在发育过程中以及兴奋性听觉神经(nVIII)输入去传入后GABA(B)R表达的分布进行了表征。在被认为先于并包括GABA能输入突触形成的发育阶段,我们在听觉脑桥中使用了一种针对大鼠GABA(B)Rs产生的多克隆抗体。早在胚胎第14天(E14),在NA、NM、NL和SON中就观察到密集标记。在更早的年龄,免疫反应性存在于胞体中,呈弥漫性染色,几乎没有点状标记。到E21时,已知听觉核团的结构和功能已成熟,GABA(B)免疫反应性的特征是在NM、NL和一部分NA神经元中有密集的点状标记,但在SON中的标记稀疏。孵化后小鸡去除耳蜗和nVIII神经元,在存活14天或28天后,免疫反应性仅略有下降,这表明大部分GABA(B)Rs可能在突触后或GABA能终末表达。我们用免疫金透射电镜证实了这一解释,在那里清楚地观察到突触后膜部位的表达。GABA(B)R分布的表征丰富了我们对这个经过充分研究的神经元回路中对中枢听觉处理的抑制性影响的完整补充的理解。