转基因NOD小鼠胰腺Kuzbanian-DN mRNA的表达定位

Expression and localization of Kuzbanian-DN mRNA in trans-gene NOD mouse pancreas cells

目的检测Kuzbanian显性失活(Kuz-DN)基因mRNA在转基因非肥胖型糖尿病(NOD)小鼠胰腺中的表达定位。方法转Kuz-DN基因的NOD小鼠8只,取胰腺,行冷冻与石蜡切片。用地高辛标记的Kuz-DN正义链及反义链cRNA探针,对组织切片进行原位杂交组织化学检测。结果在转基因阳性NOD小鼠胰腺组织中检测到强Kuz-DN mRNA信号。结论Kuz-DN mRNA在转基因动物模型定位、定性的实验结果,为Kuz基因改造后干预Notch/Delta信号传导途径,通过"旁侧抑制"影响细胞分化的基因治疗方案的研究提供了重要依据。

Objective Kuzbanian （KUZ） is the functional homologue of the vertebrate ADAM10. ADAMs are a protein family of cell surface-anchored membrane, whose members have extensive biological activities. This paper was to explore KUZ-DN （the dominant negative construct of KUZ） mRNA expression and localization in the trans-gene NOD mouse pancreas tissue and to provide molecular basis for the mechanism of the gene intervened stem cell differentiation to treat diabetes. Methods Pancreas was frozen and paraffin Sections of pancreas in eight trans-gene NOD mice were made. Non-radioactivity in situ hybridization experiment was conducted on the tissue sections by the cRNA probes of KUZ-DN （360 bp） sense and antisense with DIG labeled. Results There was the strong positive KUZ-DN mRNA signal report of in situ hybridization in the trans-gene NOD mouse pancreas cells. Sense chain probes and blank comparison sections were negative. Conclusion The positive expression and localization of KUZ-DN mRNA in the trans-gene NOD mouse pancreas cells provides a vital data for the gene therapy research project of the KUZ-DN intervened Notch/Delta signaling pathway by the stem cell differentiation to treat diabetes.