Rafaeloff R, Barlow S W, Rosenberg L, Vinik A I
Diabetes Institutes, Department of Internal Medicine, Eastern Virginia Medical School, Norfolk 23510, USA.
Diabetologia. 1995 Aug;38(8):906-13. doi: 10.1007/BF00400578.
We have reported previously that cellophane wrapping of the hamster pancreas is a stimulus that leads to the induction of duct epithelial cell proliferation, followed by endocrine cell differentiation and new islet formation. Reg is a candidate gene that has been reported to be expressed in regenerating pancreatic islets, suggesting a role in islet growth. We examined Reg gene expression in the cellophane-wrap model by isolating total RNA from hamster pancreata at various times after wrapping. Northern blot analysis using a rat cDNA Reg probe showed no expression of Reg in control non-wrapped hamster pancreas, whereas a strong signal was detected in control wrapped rat pancreas. Using reverse transcription of RNA followed by polymerase chain reaction (PCR) we amplified, isolated and sequenced a 194 base pair product which showed homology to rat Reg in both control and wrapped hamster pancreas. When the PCR product was used as a probe for Northern blot analysis, no signal was detected in control non-wrapped pancreata. In contrast, a strong signal was detected 1 and 2 days after wrapping, which then returned to basal between 4 and 6 days after wrapping. A similar temporal pattern was observed using in situ hybridization to localize the Reg gene. One- and 2-day wrapped but not control pancreas expressed Reg in acinar cells, but not in islets.(ABSTRACT TRUNCATED AT 250 WORDS)
我们之前报道过,用玻璃纸包裹仓鼠胰腺是一种刺激因素,可导致导管上皮细胞增殖,随后是内分泌细胞分化和新胰岛形成。Reg是一个候选基因,据报道在再生的胰岛中表达,提示其在胰岛生长中起作用。我们通过在包裹后不同时间从仓鼠胰腺中分离总RNA,研究了玻璃纸包裹模型中Reg基因的表达。用大鼠cDNA Reg探针进行的Northern印迹分析显示,在未包裹的对照仓鼠胰腺中未检测到Reg表达,而在包裹的对照大鼠胰腺中检测到强信号。通过RNA逆转录后进行聚合酶链反应(PCR),我们扩增、分离并测序了一个194碱基对的产物,该产物在对照和包裹的仓鼠胰腺中均与大鼠Reg具有同源性。当将PCR产物用作Northern印迹分析的探针时,在未包裹的对照胰腺中未检测到信号。相反,在包裹后1天和2天检测到强信号,然后在包裹后4至6天恢复到基础水平。使用原位杂交定位Reg基因时观察到类似的时间模式。包裹1天和2天的胰腺而非对照胰腺的腺泡细胞中表达Reg,但胰岛中不表达。(摘要截短至250字)