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Intravital imaging in a transgenic mouse model of breast cancer
Author(s) -
Resau James H,
Williams Bart,
Lindvall Charlotta,
Zylstra Cassandra,
Looyenga Brendan,
VandenBeldt Kristin,
West Rich
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.927.1
Subject(s) - transgene , genetically modified mouse , wnt signaling pathway , in vivo , carcinogenesis , breast cancer , pathology , cancer research , biology , cancer , confocal microscopy , mammary gland , confocal , fluorescence lifetime imaging microscopy , fluorescence , microbiology and biotechnology , medicine , signal transduction , biochemistry , gene , genetics , geometry , mathematics , physics , quantum mechanics
Currently there are few ways to complete intravital imaging of well characterized transgenic mouse models. We are developing an assay to monitor by fluorescence, a transgenic, murine, mammary carcinogenesis model of breast cancer that expresses the well characterized beta‐galactosidase marker. We are utilizing a method to observe the marker in cells, and will apply the method to developing lesions and histology preparations without terminating the biologic development of the tumor and hopefully treatment induced regression in order to determine efficacy. We have crossed MMTV WNT 1 mice with mice expressing the β‐galactosidase transgene under the control of a β‐catenin‐responsive promoter (Bat‐Gal). These animals have a well characterized, developmental pattern that originates in the mammary fat pad. The β‐catenin protein is stabilized in MMTV‐Wnt1‐induced mammary tumors. Building on Tung et al (2004), we quantified Wnt1‐induced mammary tumors by measuring the activity of β‐gal with DDAOG, a red fluorescent signal. Our model uses instruments ( cri‐inc.com ) designed for multispectral, in‐vivo fluorescence imaging. We un‐mix the image cubes and separate the DDAO and background components. We can quantify expression by using software programs (Kort et al, 2003). Upon sacrifice, the animals will be further characterized using spectral and confocal microscopes to develop 3D reconstructions of the tumors.