A dual-color genetically engineered mouse model for multispectral imaging of the pancreatic microenvironment

Pancreas. 2013 Aug;42(6):952-8. doi: 10.1097/MPA.0b013e31828643df.

Abstract

Objectives: To develop a mouse model for multispectral fluorescence imaging of the pancreas and pancreatic microenvironment.

Methods: Cre/loxP technology was used to develop this model. We crossed mT/mG indicator mice, engineered to constitutively express a conditional tdTomato transgene that converts to green fluorescent protein (GFP) expression after exposure to Cre recombinase, with Pdx1-Cre transgenic mice. To characterize this model for studies of pancreas biology, we performed bright light and fluorescence imaging of body cavities and intact organs and confocal microscopy of pancreata from offspring of Pdx1-Cre and mT/mG crosses.

Results: Pdx1-Cre-mT/mG mice demonstrated bright GFP expression within the pancreas and duodenum and intense tdTomato expression in all other organs. Green fluorescent protein expression was mosaic in Pdx1-Cre-mT/mG pancreata, with most showing extensive conversion from tdTomato to GFP expression within the epithelial-derived elements of the pancreatic parenchyma. Because both GFP and tdTomato are membrane targeted, individual cell borders were clearly outlined in confocal images of mT/mG pancreata.

Conclusions: This mouse model enables multispectral fluorescence imaging of individual cells and cell processes at the microscopic level of the pancreatic microenvironment; it should prove valuable for a variety of fluorescence imaging studies, ranging from pancreatic development to pancreatic cancer biology.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Diagnostic Imaging / methods
  • Disease Models, Animal
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Homeodomain Proteins / genetics
  • Homeodomain Proteins / metabolism
  • Integrases / genetics
  • Integrases / metabolism
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism*
  • Mice
  • Mice, 129 Strain
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Microscopy, Confocal
  • Pancreatic Neoplasms / diagnosis
  • Pancreatic Neoplasms / genetics
  • Pancreatic Neoplasms / metabolism*
  • Plant Proteins / genetics
  • Plant Proteins / metabolism
  • Red Fluorescent Protein
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Time Factors
  • Trans-Activators / genetics
  • Trans-Activators / metabolism
  • Tumor Microenvironment*

Substances

  • Homeodomain Proteins
  • Luminescent Proteins
  • Plant Proteins
  • Trans-Activators
  • pancreatic and duodenal homeobox 1 protein
  • Green Fluorescent Proteins
  • Cre recombinase
  • Integrases