Document Detail

Quantitative Assessment of Pdx1 Promoter Activity in Vivo Using a Secreted Luciferase Reporter System.
MedLine Citation:
PMID:  24029239     Owner:  NLM     Status:  Publisher    
The luciferase reporter system is useful for the assessment of various biological processes in vivo. The transcription factor pancreatic 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 homogenates of various organs, and immunohistochemistry revealed that GLuc expression and activity 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-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 β-cells.
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
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2013-9-12
Journal Detail:
Title:  Endocrinology     Volume:  -     ISSN:  1945-7170     ISO Abbreviation:  Endocrinology     Publication Date:  2013 Sep 
Date Detail:
Created Date:  2013-9-13     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0375040     Medline TA:  Endocrinology     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Department of Metabolic Disorders (W.N., K.E., M.K., T.N., H.U., M.H., K.Y.), Diabetes Research Center, Department of Laboratory Animal Medicine (M.G., Y.S., T.O.), and Section of Animal Models (T.O.), Department of Infectious Disease, Research Institute, National Center for Global Health and Medicine, Tokyo 162-8655, Japan; Department of Gastroenterological Surgery (K.E., T.F.), Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama 700-8558, Japan; and Department of Surgery (A.M., Y.Y.), Jichi Medical University, Tochigi 329-0498, Japan.
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