Su Y Y, Fry K R, Lam D M, Watt C B
Cell Mol Neurobiol. 1986 Dec;6(4):331-47. doi: 10.1007/BF00711404.
Enkephalin-like immunoreactive amacrine cells were visualized using the highly sensitive avidin-biotin method. The somas of these cells were situated in the inner nuclear and ganglion cell layers. Enkephalin-stained processes were observed in layers 1, 3, and 5 of the inner plexiform layer. The biosynthesis of sulfur-containing compounds in the goldfish retina was studied by means of a pulse-chase incubation with 35S-methionine. A 35S-labeled compound, which comigrated with authentic Met5-enkephalin on high-performance liquid chromatography (HPLC), was synthesized and was bound competitively by antibodies to enkephalin and by opiate receptors. This compound was tentatively identified as "Met5-enkephalin." The newly synthesized 35S-Met5-enkephalin was released upon depolarization of the retina with a high K+ concentration. This K+-stimulated release was greatly suppressed by 5 mM Co2+, suggesting that the release was Ca2+ dependent. Using a double-label technique, enkephalin immunoreactivity and gamma-aminobutyric acid (GABA) uptake were colocalized to some amacrine cells, whereas others labeled only for enkephalin or GABA. The possible significance of enkephalin-GABA interactions is also discussed.
使用高灵敏度抗生物素蛋白-生物素方法观察脑啡肽样免疫反应性无长突细胞。这些细胞的胞体位于内核层和神经节细胞层。在内网状层的第1、3和5层观察到脑啡肽染色的突起。通过用35S-甲硫氨酸进行脉冲追踪孵育研究金鱼视网膜中含硫化合物的生物合成。合成了一种在高效液相色谱(HPLC)上与真实的Met5-脑啡肽共迁移的35S标记化合物,该化合物能与脑啡肽抗体和阿片受体竞争性结合。该化合物被初步鉴定为“Met5-脑啡肽”。用高钾浓度使视网膜去极化时,新合成的35S-Met5-脑啡肽被释放。5 mM Co2+可极大地抑制这种钾离子刺激的释放,表明该释放依赖于钙离子。使用双重标记技术,脑啡肽免疫反应性和γ-氨基丁酸(GABA)摄取定位于一些无长突细胞,而其他细胞仅标记有脑啡肽或GABA。还讨论了脑啡肽-GABA相互作用的可能意义。