应用地高辛标记探针原位杂交方法检测仔猪小肠FcRn的表达  

Application of In situ Hybridization with Digoxin-labeled Probe in Expression Detection of FcRn in Small Intestine of Piglets

作　　者：陈丽颖[1] 范沛[1] 杨国宇[1] 韩立强[1] 赵瑞丽[2] 乔新安[1] 林茂旺[1] 李海霞[1] 王艳玲[1] 

机构地区：[1]河南农业大学牧医工程学院,郑州450002 [2]华南农业大学兽医学院,广州510642

出　　处：《农业生物技术学报》2008年第5期775-779,共5页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金资助项目(No.30671531)资助

摘　　要：根据GenBank公布的猪新生儿Fc受体(FcRn)重链(α链)mRNA基因保守序列,设计并合成带地高辛标记的原位杂交试验用cDNA探针。经仔猪小肠冰冻组织切片最佳厚度选择、小肠组织中内源性辣根过氧化物酶祛除等试验条件优化,建立了利用地高辛标记探针检测仔猪小肠FcRn mRNA表达的原位杂交方法。结果显示,在仔猪十二指肠、空肠和回肠组织的小肠腺和粘膜下层等部位均检测到清晰的杂交信号,表明FcRn mRNA在上述营养物质吸收部位均有表达,提示FcRn在这些部位的存在与肠道IgG转运具有相关性。表明应用地高辛标记探针进行仔猪小肠FcRn mRNA的原位杂交检测效果良好。A digoxin-labeled single-strand cDNA probe for detection of swine neonatal Fc receptor （FcRn） mRNA was plotted according to the conserved region of the gene from α chain of porcine FcP, n, and the method of in situ hybridization （ISH） was effectively established for the expression of FcRn in small intestine of piglets, which was optimized through a series of trials such as the elimination of endogenous horseradish peroxidase （HRP）, fixation agent selection. The results showed that limpid hybridization signals were detected in the intestinal glands and submucosa of duodenum, jejunum and ileum, indicating that FcRn mRNA was expressed in such nutrition-absorption tissues, which implied the correlation of the presence of FcRn and IgG transportation in the intestine. The ISH detection on the intestinal expression of swine FcRn mRNA turned out to be effective.

关 键 词：原位杂交 新生儿Fc受体(FeRn) 仔猪 小肠 

分 类 号：S188[农业科学—农业基础科学]