Ribak C E, Vaughn J E, Saito K, Barber R, Roberts E
Brain Res. 1977 Apr 22;126(1):1-18. doi: 10.1016/0006-8993(77)90211-6.
Glutamate decarboxylase (GAD), the enzyme that synthesizes the neurotransmitter gamma-aminobutyric acid (GABA), has been localized in the rat olfactory bulb by immunocytochemical methods with both light and electron microscopy. The light microscopic results demonstrated GAD-positive puncta concentrated in the external plexiform layer and in the glomeruli of the glomerular layer. In addition, GAD-positive reaction product stained the dentrites and somata of granule and periglomerular cells. The electron microscopic observations confirmed the presence of GAD-positive reaction product within granule and periglomerular somata and dendrites. In electron micrographs of the external plexiform layer, the gemmules which arise from the distal dentrites of granule cells were also observed to be filled with reaction product, and these structures corresponded in size and location to the puncta observed in light microscopic preparations. The gemmules were observed to form reciprocal dendrodentritic synaptic junctions with mitral cell dentrites which lacked reaction product. In the glomeruli, GAD-positive reaction product was observed in the dentritic shafts and gemmules of periglomerular cells which also formed reciprocal dendrodentritic synaptic contacts with mitral/tufted cell dentrites. The localization of GAD in known inhibitory neurons of the olfactory bulb supports the case that these local circuit neurons use GABA as their neurotransmitter. The present study demonstrates that GAD molecules located within certain neuronal somata and dentrites can be visualized with antisera prepared against GAD that was purified from synaptosomal fractions of mouse brains. This finding suggests that the lack of GAD staining within somata and dentrites of GABA-ergic neurons noted in previous studies of the cerebellum and spinal cord was probably due to low GAD concentrations, rather than to antigenic differences among GAD molecules located in different portions of the neuron. A striking differences among GAD molecules located in different portions of the neuron. A striking difference between the granule and periglomerular neurons of the olfactory bulb and the neurons of the cerebellum and spinal cord is that the former have presynaptic dentrites while the latter do not. Since GAD-positive reaction product can be detected in the somata and dentrites of GABA-ergic neurons which have presynaptic dentrites, it is suggested that these neurons may differ from other GABA-ergic neurons with respect to either transport or metabolism of GAD.
谷氨酸脱羧酶(GAD)是一种合成神经递质γ-氨基丁酸(GABA)的酶,通过免疫细胞化学方法,利用光学显微镜和电子显微镜已将其定位在大鼠嗅球中。光学显微镜结果显示,GAD阳性小点集中在外侧丛状层和肾小球层的肾小球中。此外,GAD阳性反应产物使颗粒细胞和球周细胞的树突和胞体着色。电子显微镜观察证实了颗粒细胞和球周细胞的胞体及树突内存在GAD阳性反应产物。在外侧丛状层的电子显微照片中,还观察到颗粒细胞远端树突产生的小芽充满了反应产物,这些结构在大小和位置上与光学显微镜标本中观察到的小点相对应。观察到小芽与缺乏反应产物的二尖瓣细胞树突形成相互的树突-树突突触连接。在肾小球中,在球周细胞的树突干和小芽中观察到GAD阳性反应产物,这些球周细胞也与二尖瓣/簇状细胞树突形成相互的树突-树突突触接触。GAD在嗅球已知抑制性神经元中的定位支持了这些局部回路神经元使用GABA作为神经递质的观点。本研究表明,用从小鼠脑突触体组分中纯化的GAD制备的抗血清可以使位于某些神经元胞体和树突内的GAD分子可视化。这一发现表明,在先前对小脑和脊髓的研究中,GABA能神经元的胞体和树突内缺乏GAD染色可能是由于GAD浓度低,而不是由于位于神经元不同部位的GAD分子之间的抗原差异。嗅球的颗粒细胞和球周神经元与小脑和脊髓的神经元之间的一个显著差异是,前者有突触前树突,而后者没有。由于在有突触前树突的GABA能神经元的胞体和树突中可以检测到GAD阳性反应产物,因此表明这些神经元在GAD的转运或代谢方面可能与其他GABA能神经元不同。