Evaluación cuantitativa del remodelado arterial a partir del análisis de imágenes de microscopía confocal de la...

The pathophysiological process of tumorigenesis involves numerous parallels with the physiological process of placentation. One critical feature that both tumors and embryos share is the need to access the host blood supply. During gestation, the uterine spiral arteries, from which maternal blood is accessed, undergo a remodeling process to become enlarged, dilated, low-resistance vessels capable of delivering an increased volume of blood through placental circulatory connections made by invasive, extravillous trophoblasts. Decidual immune cells are associated with the initial stages of this remodeling prior to the arrival of trophoblasts, and mouse studies have demonstrated that, in the absence of IFNγ produced by uterine natural killer cells, spiral artery remodeling is impaired. To better understand the role of inflammatory decidual immune cells in spiral artery remodeling, it is important to have an assay that quantifiably assesses the remodeling process. Here we describe a confocal microcopy-based methodology for digitally evaluating individual vessels within the decidua basalis of mid-gestation mouse concepti by immunofluorescence staining using DAPI for general cellularity and antibodies to CAV1 (caveolin 1) and SMA (smooth muscle alpha actin) for ECs (endothelial cells) and VSMCs (vascular smooth muscle cells), respectively. Reflecting the loss of VSMCs during normal remodeling, SMA within the vascular media is markedly reduced between gd 7.5 and 9.5, while retention of SMA is a characteristic feature of impaired remodeling. Through quantitative analysis of the relative percentage of SMA within vessel walls, the methodology described here provides an objective means to assess impairment of the spiral artery remodeling process.