Quantitative assessment of Pdx1 promoter activity in vivo using a secreted luciferase reporter system

Wataru Nishimura, Koki Eto, Atsushi Miki, Motohito Goto, Miho Kawaguchi, Takao Nammo, Haruhide Udagawa, Masaki Hiramoto, Yukiko Shimizu, Tadashi Okamura, Toshiyoshi Fujiwara, Yoshikazu Yasuda, Kazuki Yasuda

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

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 β-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 β-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 homogenatesof various organs,andimmunohistochemistryrevealed thatGLucexpressionandactivity were exponentially higher in pancreatic β-cells than in pancreatic non-β-cells, the duodenum, and other organs. In addition, GLuc activity secreted into the culture medium from islets isolated from Pdx1-GLucmicecorrelatedwiththenumberofislets. ThetransplantationofPdx1-GLucisletsintosevere 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 β-cells.

Original languageEnglish
Pages (from-to)4388-4395
Number of pages8
JournalEndocrinology
Volume154
Issue number11
DOIs
Publication statusPublished - Nov 1 2013

    Fingerprint

ASJC Scopus subject areas

  • Endocrinology

Cite this

Nishimura, W., Eto, K., Miki, A., Goto, M., Kawaguchi, M., Nammo, T., Udagawa, H., Hiramoto, M., Shimizu, Y., Okamura, T., Fujiwara, T., Yasuda, Y., & Yasuda, K. (2013). Quantitative assessment of Pdx1 promoter activity in vivo using a secreted luciferase reporter system. Endocrinology, 154(11), 4388-4395. https://doi.org/10.1210/en.2012-2248