Efficient and controlled gene expression in mouse pancreatic islets by arterial delivery of tetracycline-inducible adenoviral vectors
Efficient and controlled gene expression in mouse pancreatic islets by arterial delivery of tetracycline-inducible adenoviral vectors
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DOI：
10.1677/jme.1.02189
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发表时间：
2007-02-01
影响因子：
3.5
通讯作者：
Oka, Yoshitomo
中科院分区：
文献类型：
--
作者：
Takahashi, Rui;Ishihara, Hisamitsu;Oka, Yoshitomo
腺病毒介导的基因转移已广泛应用于胰岛研究。然而，感染效率在报告中有所不同。我们重新评估了感染效率，这里定义为小鼠胰岛中表达转基因的胰岛细胞的百分比。用腺病毒感染胰岛，感染效率为30-40%，转导细胞分布于胰岛周围。在感染前用胶原酶处理分离的胰岛使感染效率增加到70%，但抑制葡萄糖刺激的胰岛素分泌。为了探索更有效的策略，我们采用动脉递送病毒颗粒到原位胰岛。通过腹腔动脉和上级肠系膜动脉递送腺病毒（类似于每个胰腺10（8）个颗粒）是高效的，导致超过90%的转导而不损害葡萄糖刺激的胰岛素分泌。动脉递送携带甘油激酶cDNA的腺病毒使我们能够观察到甘油刺激的小鼠胰岛胰岛素分泌，这在我们采用常规方法时没有观察到。此外，动脉递送方法与四环素诱导的腺病毒系统相结合诱导有效和受控的转基因表达。我们的数据提供了新的见解基因转导方法，使用重组腺病毒在小鼠胰岛，因此预计将有助于未来的基础和临床胰岛研究应用。
Gene transfer with adenovirus vectors has been used extensively for pancreatic islet research. However, infection efficiency varies among reports. We reevaluated the infection efficiency, defined here as the percentage of islet cells expressing transgenes, in mouse islets. When the isolated islets were infected with adenoviruses, the infection efficiency was found to be 30-40% and the transduced cells were distributed in the islet periphery. Collagenase treatment of isolated islets before infection increased the infection efficiency to 70%, but with suppression of glucose-stimulated insulin secretion. To explore more efficient strategies, we employed arterial delivery of virus particles to islets in situ. Delivery of adenovirus (similar to 10(8) particles per pancreas) through the celiac and superior mesenteric arteries is highly efficient, resulting in more than 90% transduction without impairing glucose-stimulated insulin secretion. Arterial delivery of an adenovirus harboring glycerol kinase cDNA allowed us to observe glycerol-stimulated insulin secretion from mouse islets, which was not observed when we employed the conventional method. Furthermore, the arterial delivery method combined with a tetracycline-inducible adenovirus system induced efficient and controlled transgene expression. Our data provide new insights into gene transduction methods using recombinant adenoviruses in mouse islets, and are therefore anticipated to contribute to future basic and clinical islet research applications.