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目的:建立敏感的非放射性原位杂交技术检测胃肠调节肽基因的表达.方法:用体外转录法制备地高辛精(Dig)标记生长抑素cRNA探针,在4%多聚甲醛固定的胃、十二指肠和胰腺恒冷箱切片标本上进行了原位杂交,结果:杂交阳性部位呈紫蓝色,位于胞浆,背景浅淡,反差强烈.含生长抑素mRNA细胞与以前所报道的生长抑素免疫反应细胞的形态、分布均一致。结论:Dig标记cRNA探针原位杂交法敏感、简便、迅速、可靠,可用于胃肠调节肽基因表达的研究.
Objective: To establish a sensitive non-radioactive in situ hybridization to detect the expression of gastrointestinal regulator peptide gene. Methods: Digoxigenin (Dig) labeled somatostatin cRNA probe was prepared by in vitro transcription. In situ hybridization was performed on 4% paraformaldehyde fixed stomach, duodenum and pancreas cryostat sections. Results: The positive hybridization site was purple-blue, located in the cytoplasm, pale background, strong contrast. Somatostatin mRNA containing cells and previously reported somatostatin immunoreactive cells morphology, distribution are the same. Conclusion: Dig-labeled cRNA probe in situ hybridization method is sensitive, simple, rapid and reliable and can be used for the study of gastrointestinal regulator peptide gene expression.