Quantitative Assessment of Pdx1 Promoter Activity in Vivo Using a Secreted Luciferase Reporter System

被引：12

作者：

Nishimura, Wataru ^{[1
]}

Eto, Koki ^{[1
,4
]}

Miki, Atsushi

Goto, Motohito ^{[2
]}

Kawaguchi, Miho ^{[1
]}

Nammo, Takao ^{[1
]}

Udagawa, Haruhide ^{[1
]}

Hiramoto, Masaki ^{[1
]}

Shimizu, Yukiko ^{[2
]}

Okamura, Tadashi ^{[2
,3
]}

Fujiwara, Toshiyoshi ^{[4
]}

Yasuda, Yoshikazu

Yasuda, Kazuki ^{[1
]}

机构：

[1] Natl Ctr Global Hlth & Med, Dept Metab Disorders, Diabet Res Ctr, Tokyo 1628655, Japan

[2] Natl Ctr Global Hlth & Med, Dept Lab Anim Med, Tokyo 1628655, Japan

[3] Natl Ctr Global Hlth & Med, Sect Anim Models, Dept Infect Dis, Res Inst, Tokyo 1628655, Japan

[4] Okayama Univ, Dept Surg Gastroenterol, Grad Sch Med Dent & Pharmaceut Sci, Okayama 7008558, Japan

来源：

ENDOCRINOLOGY | 2013年 / 154卷 / 11期

基金：

日本学术振兴会;

关键词：

PANCREATIC BETA-CELLS; ISLET GRAFT-SURVIVAL; GAUSSIA-LUCIFERASE; GENE; MODEL; MICE; MASS; PET; TRANSPLANTATION; EXPRESSION;

D O I：

10.1210/en.2012-2248

中图分类号：

R5 [内科学];

学科分类号：

1002 ; 100201 ;

摘要：

The luciferase reporter system is useful for the assessment of various biological processes in vivo. The transcription factor pancreatic and duodenal homeobox 1 (Pdx1) is critical for the formation and the function of pancreatic beta-cells. A novel reporter system using secreted Gaussia princeps luciferase (GLuc) under the control of a Pdx1 promoter was generated and activated in rat and mouse beta-cell lines. This Pdx1-GLuc construct was used as a transgene for the generation of reporter mice to monitor Pdx1 promoter activity in vivo via the measurement of secreted GLuc activity in a small aliquot of blood. Significantly increased plasma GLuc activity was observed in Pdx1-GLuc mice. Analysis of Pdx1-GLuc mice by bioluminescence imaging, GLuc reporter assays using homogenates of various organs, and immunohistochemistry revealed that GLuc expression and activity were exponentially higher in pancreatic beta-cells than in pancreatic non-beta-cells, the duodenum, and other organs. In addition, GLuc activity secreted into the culture medium from islets isolated from Pdx1-GLuc mice correlated with the number of islets. The transplantation of Pdx1-GLuc islets into severe combined immunodeficiency mice elevated their plasma GLuc activity. Conversely, a partial pancreatectomy in Pdx1-GLuc mice reduced plasma GLuc activity. These results suggest that a secreted luciferase reporter system in vivo enables not only the monitoring of promoter activity but also a quantitative and minimally invasive assessment of physiological and pathological changes in small cell masses, such as pancreatic beta-cells.

引用

页码：4388 / 4395

页数：8