McKillop J M, Foy W L, Johnston C F, Shaw C, Murphy R F, Buchanan K D
Department of Biochemistry, Queen's University of Belfast, Northern Ireland, U.K.
Brain Res. 1988 May 3;447(2):239-45. doi: 10.1016/0006-8993(88)91125-0.
Using radioimmunoassay, reverse phase high pressure liquid chromatography (rp HPLC) and immunohistochemistry, we have identified gastrin releasing peptide-immunoreactivity (GRP-IR) in the rat retina. The concentration of GRP-IR in retinal extracts was 7.4 +/- 0.6 ng/g wet wt. (mean +/- S.E.M. n = 15). There was no significant difference between the levels of immunoreactivity in 12-h light and 12-h dark adapted retinae. rp HPLC analysis of retinal extracts demonstrated that two main immunoreactive components were present which corresponded in retention time to GRP10 (neuromedin C) and GRP14 (GRP14-27). A small amount of material also co-eluted with GRP27. Using immunohistochemistry, the immunoreactivity has been localised in the inner retinal layers. Immunoreactive somata were present in the proximal inner nuclear layer and in the ganglion cell layer. Fibre staining was present in laminae 2 and 4 of the inner plexiform layer. Somatal staining was increased by pretreatment of retinae with vincristine while the laminar staining was markedly reduced. These results demonstrate the existence of GRP-like peptides in the rat retina which has not previously been reported.
利用放射免疫分析、反相高压液相色谱法(rp HPLC)和免疫组织化学方法,我们在大鼠视网膜中鉴定出了胃泌素释放肽免疫反应性物质(GRP-IR)。视网膜提取物中GRP-IR的浓度为7.4±0.6 ng/g湿重(平均值±标准误,n = 15)。在12小时光照适应和12小时黑暗适应的视网膜中,免疫反应性水平之间没有显著差异。对视网膜提取物的rp HPLC分析表明,存在两种主要的免疫反应成分,其保留时间与GRP10(神经介素C)和GRP14(GRP14-27)相对应。还有少量物质与GRP27共洗脱。利用免疫组织化学方法,免疫反应性物质定位于视网膜内层。免疫反应性胞体存在于近端内核层和神经节细胞层。纤维染色出现在内网状层的第2层和第4层。用长春新碱预处理视网膜后,胞体染色增加,而层状染色明显减少。这些结果证明了大鼠视网膜中存在GRP样肽,这在以前尚未有过报道。