骨组织地高辛标记寡核苷酸探针原位杂交方法的建立

In Situ Hybridization with Digoxigeninlabelling Oligonucleotide: an Practical Approach to Skeletal Tissues

选用５只６周龄雄性ＳＤ大鼠双侧胫骨近端，用０．５ｍｏｌ／ＬＥＤＴＡ脱钙后，用非放射性同位素（地高辛）标记胰岛素样生长因子Ⅰ寡核苷酸探针，探讨骨组织石蜡切片原位杂交的方法。结果显示骨组织中各部位结构清晰，细胞内ｍＲＮＡ保存良好，所设对照实验证实了该方法具有较高的敏感性和特异性。表明本方法是进行骨组织的分子生物学研究一种有效。

The purpose of this study was to investigate the in situ hybridization technique of paraffinembeded skeletal tissues by nonradioactive labelling oligonucleotide, Five sixweekold male SD rats were selected and the proximal epiphyseal growth plates of bilateral tibiae were removed to be decalcified in 0.5 mol/L EDTA. The tissues were embeded by paraffin and the sections were treated and hybridized with RNasefree reagents. Control groups were set to testfied the sensitivity and the specificity of the experiment. The results showed that this method preserved the integrate structures and effectively detected the mRNA of the tissues. These indicated that it was an efficient and practical approach to the biomolecular research of skeletal tissue.