摘要
目的:观察神经元限制性沉默因子(NRSF)在正常成年小鼠胰腺组织中的表达情况。方法:以6～8周BALB/c小鼠胰腺为实验材料,制备冰冻切片,与地高辛标记的NRSF cDNA探针进行原位杂交,观察mRNA表达,并结合免疫组织化学方法检测NRSF和胰岛素的表达。结果:原位杂交显示,NRSF mRNA仅表达于胰腺组织外分泌部腺泡腺细胞中,胞浆呈蓝紫色,与免疫荧光组织化学检测NRSF蛋白表达的部位一致,而胰岛细胞中无NRSF mRNA及蛋白的表达。免疫酶组织化学染色显示,胰岛大部分细胞中表达胰岛素,胞浆染成黄棕色,而腺泡腺细胞则不表达胰岛素。结论:NRSF与胰岛素不存在共定位关系,即成年小鼠胰岛细胞不表达NRSF,而表达胰岛素。提示NRSF蛋白表达的消失可能是建立完全分化成熟、具有完好分泌反应的胰岛细胞所必需的。
Objective： To observe the mRNA and protein expression of neuron-restrictive silencer factor（NRSF） in pancreatic islet of adult mice.Methods： Frozen sections were prepared from 6～8 weeks BALB / c mouse pancreas.Tissue sections were then detected by using in situ hybridization with digoxin-labeled NRSF cDNA probes.Im-munohistochemistry methods were used to detect the protein expression of NRSF and insulin in pancreatic tissue.Results： In situ hybridization detection results demonstrated that NRSF mRNA was located in the cytoplasm of pancreatic exocrine cells.Combining with immunofluorescence detection using anti-NRSF antibody,we found that pancreatic islet cells showed no expression of NRSF mRNA and protein.The islet cells were proved by insulin staining.Conclusion： These results showed that NRSF doesn＇t colocalize with insulin in pancreatic islets of adult mice,which suggest that the disappearence of NRSF might be necessary for pancreatic islet cells maturation.
出处
《生物技术通讯》
CAS
2011年第5期608-611,共4页
Letters in Biotechnology
基金
北京市自然科学基金(5102036)
