地高辛标记反义RNA探针对大鼠不同组织生长抑素mRNA原位杂交的实验研究  

In situ hybridization for detection of somatostatin mRNA in rat tissues with digoxigenin-labelled antisense RNA probe

作　　者：张钦宪[1] 宗安民[1] 史学义[1] 乐晓萍[1] 

机构地区：[1]河南医科大学组织胚胎学教研室

出　　处：《河南医科大学学报》1994年第4期283-286,共4页Journal of Henan Medical University

基　　金：国家自然科学基金

摘　　要：应用地高辛（ＤＩＧ）标记的生长抑素（ＳＯＭ）反义ＲＮＡ探针，与Ｗｉｓｔａｒ大鼠胃、十二指肠和甲状腺组织进行原位分子杂交，以显示不同组织中ＳＯＭｍＲＮＡ对ＳＯＭ细胞进行分子水平的细胞定位和形态学观察。结果显示：杂交阳性信号呈蓝色位于胞质内，背景清晰，细胞轮廓清楚，主要分布于胃粘膜底部，十二指肠阳性细胞较少，在甲状腺滤泡旁和滤泡上皮细胞之间均可见到形态不一的杂交阳性细胞。用ＲＮａｓｅＡ预处理标本和免去探针均无杂交信号出现。表明ＤＩＧ标记的反义ＲＮＡ探针具有高度的敏感性和特异性。In situ hybridization with digoxigenin-labelled antisense RNA probes prepared with sp6 transcription system in vitro was used to detect the somatostatin mRNA in the endocrine cells of the gastric antrum,the duodenum and the thyroid gland of Wistar rats.The results demonstrated that the positive signal of hybridization was blue and was located in the cytoplasm,the background was clear after 4 hours staining developed and light violet,after 16 hours.No positive signal was found in cell nuclei and on both samples pretreated with RNase A and the samples treated without probes.The positive cells in the gastric antrum were mainly located at the lower portion of gastric mucosa.They were ovoid or fusiform and occasionally with a thin,long process.In the duodenum there were few positive cells.The positive cells in the thyroid gland were distributed between the follicles or between epithelial cells.They were oviod in shape,The results indicated that the hybridization signal with digoxigenin-labelled antisense RNA probes showed high resolution and sensitivity with less background,and the experimental cycles were shorter(3-4 days) than that with the radio-labelled probe.Besides,the localization of the somatostatin cells at molecular level in the gastric antrum,duodenum and thyroid gland could be performed.

关 键 词：原位杂交 RNA 探针 地高辛 生长抑素 

分 类 号：R394[医药卫生—医学遗传学] Q343[医药卫生—基础医学]