Yazulla S, Studholme K M, Vitorica J, de Blas A L
Department of Neurobiology and Behavior, State University of New York, Stony Brook 11794.
J Comp Neurol. 1989 Feb 1;280(1):15-26. doi: 10.1002/cne.902800103.
A monoclonal antibody (mAb 62-3G1) to the GABAA receptor/benzodiazepine receptor/Cl- channel complex from bovine brain was used with light and electron microscopy in goldfish retina and light microscopy in chicken retina to localize GABAA receptor immunoreactivity (GABAr-IR). GABAr-IR was found in the outer plexiform layer (OPL) in both species, in three broad bands in the inner plexiform layer (IPL) of goldfish, and in seven major bands of the chicken IPL. A small percentage of amacrine cell bodies (composing at least three types) were stained in chicken. In goldfish OPL, GABAr-IR was localized intracellularly and along the plasma membrane of cone pedicles, whereas rod spherules were lightly stained, but always only intracellularly. In chicken, all three sublayers of the OPL were GABAr-IR. The presence of GABAr-IR on photoreceptor terminals is consistent with data indicating feedback from GABAergic horizontal cells to cones. In the goldfish IPL, GABAr-IR was localized to postsynaptic sites of amacrine cell synapses; intracellular staining of processes in the IPL also was observed in presumed "GABAergic" targets. A comparison of GABAr-IR with the distributions of 3H-muscimol uptake/binding, glutamate decarboxylase-IR, GABA-IR, and 3H-GABA uptake in the IPL showed either a reasonable correspondence or mismatch, depending on the marker, species, and lamina within the IPL. The distribution of GABAr-IR in the retina corresponded better with the 3H-muscimol than with 3H-benzodiazepine binding patterns yet overall was in excellent agreement with many other physiological and anatomical indicators of GABAergic function. We suggest that intracellular GABAr-IR represents the biosynthetic and/or degradative pathway of the receptor and we conclude that mAb 62-3G1 is a valid marker of GABAA receptors in these retinas and will serve as a useful probe with which to address the issue of mismatches between the localization of GABAA receptors and indicators of presynaptic GABAergic terminals.
一种针对牛脑γ-氨基丁酸A型受体/苯二氮䓬受体/氯离子通道复合物的单克隆抗体(mAb 62 - 3G1)被用于金鱼视网膜的光镜和电镜研究以及鸡视网膜的光镜研究,以定位γ-氨基丁酸A型受体免疫反应性(GABAr - IR)。在这两个物种的视网膜中,外网状层(OPL)均发现有GABAr - IR;在金鱼的内网状层(IPL)中有三条宽带状区域存在GABAr - IR,而在鸡的IPL中有七条主要带状区域存在GABAr - IR。在鸡的视网膜中,一小部分无长突细胞胞体(至少由三种类型组成)被染色。在金鱼的OPL中,GABAr - IR定位于细胞内以及视锥小足的质膜上,而视杆小球仅有轻微染色,且总是仅在细胞内。在鸡的视网膜中,OPL的所有三个亚层均有GABAr - IR。光感受器终末上存在GABAr - IR与表明γ-氨基丁酸能水平细胞向视锥细胞反馈的数据一致。在金鱼的IPL中,GABAr - IR定位于无长突细胞突触的突触后位点;在假定的“γ-氨基丁酸能”靶点中也观察到IPL中突起的细胞内染色。将IPL中GABAr - IR的分布与³H - 蝇蕈醇摄取/结合、谷氨酸脱羧酶免疫反应性、γ-氨基丁酸免疫反应性以及³H - γ-氨基丁酸摄取的分布进行比较,结果显示根据标记物、物种以及IPL内的层不同,存在合理的对应或不匹配情况。视网膜中GABAr - IR的分布与³H - 蝇蕈醇的结合模式比与³H - 苯二氮䓬的结合模式更相符,但总体上与γ-氨基丁酸能功能的许多其他生理和解剖学指标高度一致。我们认为细胞内GABAr - IR代表了受体的生物合成和/或降解途径,并且我们得出结论,mAb 62 - 3G1是这些视网膜中γ-氨基丁酸A型受体的有效标记物,将作为一个有用的探针来解决γ-氨基丁酸A型受体定位与突触前γ-氨基丁酸能终末指标之间的不匹配问题。