Abstract
We have developed a transgenic green fluorescent protein (GFP) nude mouse with ubiquitous GFP expression. The GFP nude mouse was obtained by crossing nontransgenic nude mice with the transgenic C57/B6 mouse in which the β-actin promoter drives GFP expression in essentially all tissues. In the adult mice, many organs brightly expressed GFP, including the spleen, heart, lungs, spleen, pancreas, esophagus, stomach, and duodenum as well as the circulatory system. The liver expressed GFP at a lesser level. The red fluorescent protein (RFP) transgenic nude mouse was obtained by crossing non-transgenic nude mice with the transgenic C57/B6 mouse in which the beta-actin promoter drives RFP (DsRed2) expression in essentially all tissues. In the RFP nude mouse, the organs all brightly expressed RFP, including the heart, lungs, spleen, pancreas, esophagus, stomach, liver, duodenum, the male and female reproductive systems; brain and spinal cord; and the circulatory system, including the heart, and major arteries and veins. The skinned skeleton highly expressed RFP. The bone marrow and spleen cells were also RFP positive. The cyan fluorescent protein (CFP) nude mouse was developed by crossing nontransgenic nude mice with the transgenic CK/ECFP mouse in which the β-actin promoter drives expression of CFP in almost all tissues. In the CFP nude mice, the pancreas and reproductive organs displayed the strongest fluorescence signals of all internal organs, which vary in intensity. The GFP, RFP, and CFP nude mice when transplanted with cancer cells of another color are powerful models for color-coded imaging of the tumor microenvironment (TME) at the cellular level.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Fukumura D, Xavier R, Sugiura T, Chen Y, Park EC, Lu N, Selig M, Nielsen G, Taksir T, Jain RK, Seed B (1998) Tumor induction of VEGF promoter activity in stromal cells. Cell 94:715–725
Brown EB, Campbell RB, Tsuzuki Y, Xu L, Carmeliet P, Fukumura D, Jain RK (2001) In vivo measurement of gene expression, angiogenesis and physiological function in tumors using multiphoton laser scanning microscopy. Nat Med 7:864–868
Okabe M, Ikawa M, Kominami K, Nakanishi T, Nishimune Y (1997) “Green mice” as a source of ubiquitous green cells. FEBS Lett 407:313–319
Vintersten K, Monetti C, Gertsenstein M, Zhang P, Laszlo L, Biechele S, Nagy A (2004) Mouse in red: red fluorescent protein expression in mouse ES cells, embryos, and adult animals. Genesis 40:241–246
Hadjantonakis AK, Macmaster S, Nagy A (2002) Embryonic stem cells and mice expressing different GFP variants for multiple non-invasive reporter usage within a single animal. BMC Biotechnol 2:11
Tran Cao HS, Reynoso J, Yang M, Kimura H, Kaushal S, Snyder CS, Hoffman RM, Bouvet M (2009) Development of the transgenic cyan fluorescent protein (CFP)-expressing nude mouse for “Technicolor” cancer imaging. J Cell Biochem 107:328–334
Hoffman RM (2005) The multiple uses of fluorescent proteins to visualize cancer in vivo. Nat Rev Cancer 5:796–806
Hoffman RM, Yang M (2006) Subcellular imaging in the live mouse. Nat Protoc 1:775–782
Hoffman RM, Yang M (2006) Color-coded fluorescence imaging of tumor–host interactions. Nat Protoc 1:928–935
Hoffman RM, Yang M (2006) Whole-body imaging with fluorescent proteins. Nat Protoc 1:1429–1438
Yamauchi K, Yang M, Jiang P, Xu M, Yamamoto N, Tsuchiya H, Tomita K, Moossa AR, Bouvet M, Hoffman RM (2006) Development of real-time subcellular dynamic multicolor imaging of cancer-cell trafficking in live mice with a variable-magnification whole-mouse imaging system. Cancer Res 66:4208–4214
Yang M, Jiang P, Hoffman RM (2007) Whole-body subcellular multicolor imaging of tumor-host interaction and drug response in real time. Cancer Res 67:5195–5199
Uchugonova A, Zhao M, Weinigel M, Zhang Y, Bouvet M, Hoffman RM, Koenig K (2013) Multiphoton tomography visualizes collagen fibers in the tumor microenvironment that maintain cancer-cell anchorage and shape. J Cell Biochem 114:99–102
Yang M, Reynoso J, Jiang P, Li L, Moossa AR, Hoffman RM (2004) Transgenic nude mouse with ubiquitous green fluorescent protein expression as a host for human tumors. Cancer Res 64:8651–8656
Yang M, Reynoso J, Bouvet M, Hoffman RM (2009) A transgenic red fluorescent protein-expressing nude mouse for color-coded imaging of the tumor microenvironment. J Cell Biochem 106:279–284
Suetsugu A, Osawa Y, Nagaki M, Saji S, Moriwaki H, Bouvet M, Hoffman RM (2011) Imaging the recruitment of cancer-associated fibroblasts by liver-metastatic colon cancer. J Cell Biochem 112:949–953
Suetsugu A, Hassanein MK, Reynoso J, Osawa Y, Nagaki M, Moriwaki H, Saji S, Bouvet M, Hoffman RM (2012) The cyan fluorescent protein nude mouse as a host for multicolor-coded imaging models of primary and metastatic tumor microenvironments. Anticancer Res 32:31–38
Hoffman RM (2012) In vivo cellular imaging using fluorescent proteins: methods and protocols. Methods in molecular biology 872. Springer, New York
Kocher B, Piwnica-Worms D (2013) lluminating cancer systems with genetically engineered mouse models and coupled luciferase reporters in vivo. Cancer Disc 3:616–629
Jain RK, Munn LL, Fukumura D (2002) Dissecting tumour pathophysiology using intravital microscopy. Nat Rev Cancer 2:266–276
Ritsma L, Steller EJ, Beerling E, Loomans CJ, Zomer A, Gerlach C, Vrisekoop N, Seinstra D, van Gurp L, Schäfer R, Raats DA, de Graaff A, Schumacher TN, de Koning EJ, Rinkes IH, Kranenburg O, van Rheenen J (2012) Intravital microscopy through an abdominal imaging window reveals a pre-micrometastasis stage during liver metastasis. Sci Transl Med 4:158ra145
Yang M, Li L, Jiang P, Xu M, Tsuchiya H, Tomita K, Moossa AR, Hoffman RM (2003) Dual-color fluorescence imaging distinguishes tumor cells from induced host angiogenic vessels and stromal cells. Proc Natl Acad Sci U S A 100:14259–14262
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media New York
About this protocol
Cite this protocol
Hoffman, R.M. (2014). Transgenic Nude Mice Ubiquitously Expressing Fluorescent Proteins for Color-Coded Imaging of the Tumor Microenvironment. In: Singh, S., Coppola, V. (eds) Mouse Genetics. Methods in Molecular Biology, vol 1194. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1215-5_20
Download citation
DOI: https://doi.org/10.1007/978-1-4939-1215-5_20
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-1214-8
Online ISBN: 978-1-4939-1215-5
eBook Packages: Springer Protocols