title: Efficient and controlled gene expression in mouse pancreatic islets by arterial delivery of tetracycline-inducible adenoviral vectors.
abstract: 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 (approximately 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.
authors.affiliation: Division of Molecular Metabolism and Diabetes, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Aoba-ku, Sendai, Miyagi 980-8575, Japan.
authors.forename: Rui
authors.initials: R
authors.lastname: Takahashi
authors.forename: Hisamitsu
authors.initials: H
authors.lastname: Ishihara
authors.forename: Kazuma
authors.initials: K
authors.lastname: Takahashi
authors.forename: Akira
authors.initials: A
authors.lastname: Tamura
authors.forename: Suguru
authors.initials: S
authors.lastname: Yamaguchi
authors.forename: Takahiro
authors.initials: T
authors.lastname: Yamada
authors.forename: Hideki
authors.initials: H
authors.lastname: Katagiri
authors.forename: Yoshitomo
authors.initials: Y
authors.lastname: Oka
mesh_terms: D000256:Adenoviridae; D000818:Animals; D000900:Anti-Bacterial Agents; D005786:Gene Expression Regulation / Q000502:physiology*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000008:administration & dosage; D007261:Infusions, Intra-Arterial; D007515:Islets of Langerhans / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D013752:Tetracycline
chemical_list: D000900:Anti-Bacterial Agents; D013752:Tetracycline