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Development of a novel in situ model for visualizing murine endometrial vasculature using fluorescence intravital microscopy
Author(s) -
Leonard Sean,
Murant Coral
Publication year - 2006
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.20.4.a713-c
Subject(s) - intravital microscopy , fluorescence microscope , confocal microscopy , biology , endothelium , pathology , chemistry , microbiology and biotechnology , anatomy , in vivo , fluorescence , medicine , endocrinology , physics , quantum mechanics
Uterine vascular modification is essential for a healthy pregnancy. However, the mechanisms underlying this modification are yet to be elucidated, and there is a significant lack of functional data regarding these transient vessels. Functional quantification such as vasoreactivity, permeability parameters and blood flow dynamics is crucial to understanding these unique vessels and requires clearly defined vessel wall identification. Therefore, we developed an in situ murine model using intravital microscopy to observe endometrial blood vessels over the period of modification. Mice were anaesthetized and the pregnant uterus (gestation day 8–12) exteriorized. The anti‐mesometrial uterine wall was incised and retracted and the fetal tissue removed. The placenta was then placed fetal‐side down and superfused with physiological buffered‐superfusate. Endometrial vasculature was visualized via transillumination or fluorescence microscopy using fluorescent markers for membrane expressed glycoproteins on endothelial cells using 0.1M Lycopersicon (FITC) or 0.067M Isolectin (Alexa‐Fluor 488). Transillumination allowed for visualization of superficial vessels with limited resolution. Isolectin and Lycopersicon staining were successful in fluorescing the endothelium. There was increased background fluorescence with Lycopersicon which was not seen with Isolectin. We have successfully developed an in situ preparation for visualizing endometrial vasculature. Isolectin provides ideal staining resolution for functional characterization of these unique vessels. This work was supported by CIHR and NSERC.