Varndell I M, Bishop A E, Sikri K L, Uttenthal L O, Bloom S R, Polak J M
J Histochem Cytochem. 1985 Oct;33(10):1080-6. doi: 10.1177/33.10.3900195.
The distribution of peptide immunoreactivities predicted from the sequence of the human preproglucagon gene in enteroglucagon (EG; glicentin-like immunoreactant-containing) cells of the human gut and A cells of the pancreas has been determined by light and electron microscopic immunocytochemistry. At light microscopy the application of peroxidase-antiperoxidase and immunogold-silver staining methods has revealed that glucagon-like peptide (GLP-1 and GLP-2) immunoreactivities coexist with a glicentin-related immunodeterminant in human colorectal EG cells and pancreatic A cells. Using single and double colloidal gold probe electron immunocytochemistry, we have been able to show the coexistence of glicentin, GLP-1, and GLP-2 immunoreactivities within single EG cell secretory granules. No morphologic segregation of the proglucagon immunoreactants was observed in EG cells of the colonic mucosa. In pancreatic A cells we have localized GLP-1, GLP-2, and glucagon-[16-29] immunoreactivities solely to the electron-dense core of the secretory granules, whereas glicentin-related immunoreactivity was restricted to the electron-lucent halo. The results obtained in the present study have shown that the peptide immunoreactivities predicted from cDNA sequencing of the human preproglucagon gene are indeed expressed in colorectal EG and pancreatic A cells. The topographical segregation of immunoreactivities in the A cell secretory granule shows that antigenic determinants derived from the C-terminal portion of proglucagon are stored with glucagon in the core of the secretory granule.
通过光学显微镜和电子显微镜免疫细胞化学技术,已确定人胰高血糖素原基因序列预测的肽免疫反应性在人肠道的肠高血糖素(EG;含胰高血糖素样免疫反应物)细胞和胰腺A细胞中的分布。在光学显微镜下,过氧化物酶-抗过氧化物酶和免疫金银染色方法的应用表明,胰高血糖素样肽(GLP-1和GLP-2)免疫反应性与人结肠直肠EG细胞和胰腺A细胞中的胰高血糖素相关免疫决定簇共存。使用单胶体金探针和双胶体金探针电子免疫细胞化学技术,我们能够显示在单个EG细胞分泌颗粒中胰高血糖素、GLP-1和GLP-2免疫反应性共存。在结肠黏膜的EG细胞中未观察到胰高血糖素原免疫反应物的形态学分离。在胰腺A细胞中,我们仅将GLP-1、GLP-2和胰高血糖素-[16-29]免疫反应性定位到分泌颗粒的电子致密核心,而胰高血糖素相关免疫反应性局限于电子透明晕。本研究获得的结果表明,从人胰高血糖素原基因cDNA测序预测的肽免疫反应性确实在结肠直肠EG细胞和胰腺A细胞中表达。A细胞分泌颗粒中免疫反应性的拓扑分离表明,源自胰高血糖素原C末端部分的抗原决定簇与胰高血糖素一起储存在分泌颗粒的核心中。