In Utero Assessment of Cardiovascular Function in the Embryonic Mouse Heart Using High-Resolution Ultrasound Biomicroscopy

Abstract

Murine models are currently the preferred approach for studying the molecular mechanisms of cardiac dysfunction resulting from changes in gene expression. Transgenic and gene-targeting methods can be used to generate mice with altered cardiac size and function, and as a result, in vivo techniques are indispensible in evaluating cardiac phenotype. Traditionally, the pathologic assessment of sacrificed hearts was used to study cardiac pathophysiology in small animals. Below we describe the use of ultrasound biomicroscopy-Doppler analysis to temporally assess cardiac function in mouse embryos. Methods are described for obtaining 2D, pulsed-wave Doppler, and M-mode imaging using standard clinical cardiac ultrasound imaging planes.

Bone Marrow Progenitor Cell Therapy-Mediated Paracrine Regulation of Cardiac miRNA-155 Modulates Fibrotic Response in Diabetic Hearts

“…CFs were then trypsinized (TrypLE™ Express, Gibco®) and replated on tissue culture grade dishes in DMEM/M199 medium and maintained at 37°C in humid air with 5% CO 2 . Confluent cells were almost exclusively fibroblasts (>98% purity), as previously reported [20], [21]. The cells were morphologically homogeneous with typical bipolar configuration observed by inverse microscopy.…”

“…Cardiac fibroblasts (CFs) were isolated from hearts of 12-week-old C57BLKS/J mice, as previously described [20], [21]. Briefly, the explanted hearts from mice were perfused with PBS-EDTA for 10 minutes, cut into pieces and digested with enzyme solution containing collagenase type II, trypsin and α-chymotrypsin (Worthington Biochem., Lakewood, NJ) at 37°C for 1 h, until the tissue blocks disappeared.…”

Use of Real-Time Ultrasonography as an Alternative Method for Early Detection, Confirmation and Evaluation of Rat Pregnancy

“…Because of advances in ultrasound imaging technology, commercially available ultrasound systems now have the spatial and temporal resolution to obtain accurate images of rat and mouse hearts, kidneys and other target tissues (Coatney 2001;Golden et al 2012;Stypmann 2007). The primary aim of this study was to evaluate the use of real-time ultrasound examination in the accurate detection of rat pregnancy.…”

“…Until recently, the use of ultrasonography was limited to larger non-rodent species (Beccaglia and Luvoni 2006;Carr et al 2012;Lenard et al 2007;Lindahl 1966;Quintela et al 2012;Zambelli and Prati 2006). Because of advances in ultrasound imaging technology, commercially available ultrasound systems now have the spatial and temporal resolution to obtain accurate images of rat and mouse hearts, kidneys and other organs (Coatney 2001;Golden et al 2012;Kobayashi et al 2001;Srinivasan et al 1998;Stypmann 2007). For this reason, ultrasound is a practical method for the diagnosis of pregnancy in agouti mice (Sousa et al 2012), staging of gestation and monitoring of intrauterine growth in rats (Brown et al 2006;Mu et al 2008;Nguyen et al 2012;Pallares and Gonzalez-Bulnes 2009;Ypsilantis et al 2009).…”

Preclinical Imaging: an Essential Ally in Modern Biosciences

“…Ultrasound biomicroscopy operates at higher frequencies (typically 40-200 MHz) [105,110] and is able to acquire images using 2D B-mode, pulsed and continuous wave Doppler and color flow Doppler [50,110]. This imaging modality has been applied in different contexts, namely the assessment of embryonic and eye development [111,112], cardiology [113], and oncology [114].…”