Wu Kathy H C, Penfold Philip L, Billson Francis A
Department of Clinical Ophthalmology and Save Sight Institute, University of Sydney, Sydney, New South Wales, Australia.
Clin Exp Ophthalmol. 2002 Jun;30(3):200-7. doi: 10.1046/j.1442-9071.2002.00515.x.
Glial fibrillary acidic protein (GFAP) is an established marker of retinal glia and has been shown to be modulated by several cytokines and retinal pathology. The influence of a number of factors, including post-mortem delay, storage duration and retinal pathology, on the distribution and morphology of macroglia and GFAP antigenicity was examined in human retina. The effects of these parameters on GFAP expression were estimated using immunohistochemistry, confocal microscopy and image analysis. Changes in expression of antigenicity were analysed in human retinal cryosections at three levels: constitutive,aberrant and total. The results indicated that short-term and long-term storage duration had no significant effect on GFAP immunoreactivity at all three levels of expression (P > 0.2).However, a significant increase in GFAP immunoreactivity and distribution at all three levels of expression was associated with prolonged post-mortem delay (> 30 h) (P < 0.05). This study highlights the importance of rigorous matching of post-mortem delay between control specimens in histological studies of human retinae. The study further demonstrates the utility of Eye Bank retinae fixed and stored in 2% paraformaldehyde, provided that appropriate controls are applied.
胶质纤维酸性蛋白(GFAP)是视网膜神经胶质细胞的一种既定标志物,并且已被证明会受到多种细胞因子和视网膜病变的调节。在人类视网膜中,研究了包括死后延迟、储存时间和视网膜病变在内的多种因素对大胶质细胞的分布和形态以及GFAP抗原性的影响。使用免疫组织化学、共聚焦显微镜和图像分析评估这些参数对GFAP表达的影响。在人类视网膜冰冻切片中,从组成性、异常性和总体三个水平分析抗原性表达的变化。结果表明,短期和长期储存时间对所有三个表达水平的GFAP免疫反应性均无显著影响(P>0.2)。然而,死后延迟延长(>30小时)与所有三个表达水平的GFAP免疫反应性和分布显著增加相关(P<0.05)。本研究强调了在人类视网膜组织学研究中,对照标本之间严格匹配死后延迟的重要性。该研究进一步证明了眼库中固定并储存在2%多聚甲醛中的视网膜的实用性,前提是应用适当的对照。