地高辛精标记cRNA探针原位杂交法检测大鼠胃肠胰系统生长抑素mRNA

Detection of somatostatin mRNA in rat gastroenteropancreatic sys-tem with digoxigenin-labelled cRNA probe in situ hybridization

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, nonradioactive in situ hybridization method to detect the ex-pression of gut regulatory peptides, Methods:the Dig-labelled somatostatin (SS) RNA probes were synthe-sized with the in vitro transcription system and the in situ hybridization was conducted on the rat stomach,duodenum and pancreatic cryostat sections.Results: positive reaction sites were purple-blue and located in the cytoplasm with a light background but a strong contrast.Both the morphosis and distribution of the SS mRNA positive cells corresponded well with those of the previous reported SS immunoreactive cells. Conclu-sion:the Dlg-labelled cRNA probe in situ hybridization method is simple,sensitive and reliable for the study of gut regulatory peptide gene expression.