Niedzielski A S, Wenthold R J
Laboratory of Neurochemistry, NIDCD, NIH, Bethesda, Maryland 20892.
J Neurosci. 1995 Mar;15(3 Pt 2):2338-53. doi: 10.1523/JNEUROSCI.15-03-02338.1995.
Glutamate is believed to be the principal afferent neurotransmitter in the peripheral auditory and vestibular systems. In this report, we present a comprehensive molecular analysis of ionotropic glutamate receptor gene expression in the cochlear and vestibular ganglia of the rat. Fourteen glutamate receptor subunits were studied: GluR1-4 (including flip and flop variants), GluR5-7, KA1&2, NR1, and NR2A-D. Reverse transcription of RNA followed by DNA amplification with the polymerase chain reaction was used for the initial analysis. Immunocytochemistry and in situ hybridization with subunit-specific oligonucleotides were subsequently used for cellular localization of receptor expression. AMPA (GluR2-4), kainate (GluR5&6 and KA1&2), and NMDA receptor (NR1 and NR2A-D) subunit expression was detected. Based on the relative amounts of mRNA detected by in situ hybridization, the predominant receptors expressed by cochlear and vestibular ganglion cells appear to be GluR2, GluR3, GluR4, GluR5, and NR1. At a moderate level were GluR6, NR2B, and NR2D. KA1, KA2, NR2A, and NR2C mRNAs were also expressed in ganglion cells, but at lower levels. Only the AMPA receptor subunit GluR1 and the kainate receptor subunit GluR7 were not found to be expressed in vestibulocochlear neurons. These studies suggest that functional AMPA, kainate, and NMDA receptors are present at the hair cell/vestibulocochlear nerve synapse.
谷氨酸被认为是外周听觉和前庭系统中的主要传入神经递质。在本报告中,我们对大鼠耳蜗和前庭神经节中离子型谷氨酸受体基因表达进行了全面的分子分析。研究了14种谷氨酸受体亚基:GluR1 - 4(包括翻转和翻转变体)、GluR5 - 7、KA1&2、NR1和NR2A - D。最初的分析采用RNA逆转录,随后用聚合酶链反应进行DNA扩增。免疫细胞化学和与亚基特异性寡核苷酸的原位杂交随后用于受体表达的细胞定位。检测到AMPA(GluR2 - 4)、海人藻酸(GluR5&6和KA1&2)和NMDA受体(NR1和NR2A - D)亚基的表达。根据原位杂交检测到的mRNA相对量,耳蜗和前庭神经节细胞表达的主要受体似乎是GluR2、GluR3、GluR4、GluR5和NR1。中等水平表达的是GluR6、NR2B和NR2D。KA1、KA2、NR2A和NR2C mRNA也在神经节细胞中表达,但水平较低。仅发现AMPA受体亚基GluR1和海人藻酸受体亚基GluR7在前庭耳蜗神经元中未表达。这些研究表明,功能性AMPA、海人藻酸和NMDA受体存在于毛细胞/前庭耳蜗神经突触处。