Laser-scanning velocimetry: A confocal microscopy method for quantitative measurement of cardiovascular performance in zebrafish embryos and larvae

Malone, Michael H.; Sciaky, Noah; Stalheim, Lisa; Hahn, Klaus M.; Linney, Elwood; Johnson, Gary L.

doi:10.1186/1472-6750-7-40

Cited by 59 publications

(55 citation statements)

References 32 publications

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“…As demonstrated, the flow velocity has a maximum in the center of the vessel for both diastolic and systolic flow and the flow velocities are close to zero near the vessel walls. The parabolic profile fits the prediction of the theoretical calculation of flow velocity in a cylindrical tube (Chien et al, 1977) and the values obtained here are comparable to other reports (Malone et al, 2007). The figures also suggest that the velocities of both systolic and diastolic flow in the dorsal aorta are faster than those in the cardinal vein.…”

Section: Blood Flow Measurement In Live Zebrafish Embryossupporting

confidence: 90%

Probing events with single molecule sensitivity in zebrafish and Drosophila embryos by fluorescence correlation spectroscopy

Shi

Teo

Pan

et al. 2009

Developmental Dynamics

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Zebrafish and Drosophila are animal models widely used in developmental biology. High-resolution microscopy and live imaging techniques have allowed the investigation of biological processes down to the cellular level in these models. Here, using fluorescence correlation spectroscopy (FCS), we show that even processes on a molecular level can be studied in these embryos. The two animal models provide different advantages and challenges. We first characterize their autofluorescence pattern and determine usable penetration depth for FCS especially in the case of zebrafish, where tissue thickness is an issue. Next, the applicability of FCS to study molecular processes is shown by the determination of blood flow velocities with high spatial resolution and the determination of diffusion coefficients of cytosolic and membrane-bound enhanced green fluorescent protein-labeled proteins in different cell types. This work provides an approach to study molecular processes in vivo and opens up the possibility to relate these molecular processes to developmental biology questions. Developmental Dynamics 238:3156-3167,

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Section: Blood Flow Measurement In Live Zebrafish Embryossupporting

confidence: 90%

Probing events with single molecule sensitivity in zebrafish and Drosophila embryos by fluorescence correlation spectroscopy

Shi

Teo

Pan

et al. 2009

Developmental Dynamics

View full text Add to dashboard Cite

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“…This constellation of distinctive pathologic findings in animal models [14-19, 75] has made understanding CCM pathophysiology difficult. Consequently, uncovering the molecular etiology of CCM and linking the pathogenesis in humans and animal models have proven daunting.…”

Section: Resultsmentioning

confidence: 99%

The Cardiovascular Triad of Dysfunctional Angiogenesis

Zhang

Carr

Badr

2011

Transl. Stroke Res.

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Cerebral cavernous malformation is a clinically well-defined microvascular disorder predisposing to stroke; however, the major phenotype observed in zebrafish is the cardiac defect, specifically an enlarged heart. Less effort has been made to explore this phenotypic discrepancy between human and zebrafish. Given the fact that the gene products from Ccm1/Ccm2 are nearly identical between the two species, the common sense has dictated that the zebrafish animal model would provide a great opportunity to dissect the detailed molecular function of Ccm1/Ccm2 during angiogenesis. We recently reported on the cellular role of the Ccm1 gene in biochemical processes that permit proper angiogenic microvascular development in the zebrafish model. In the course of this experimentation, we encountered a vast amount of recent research on the relationship between dysfunctional angiogenesis and cardiovascular defects in zebrafish. Here we compile the findings of our research with the most recent contributions in this field and glean conclusions about the effect of defective angiogenesis on the developing cardiovascular system. Our conclusion also serves as a bridge for the phenotypic discrepancy between humans and animal models, which might provide some insights into future translational research on human stroke.

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“…The zebrafish embryo is a common animal model to study human cardiovascular disease [30] due to its known genome [8], optical transparency [36] and rapid growth rate [25]. During early cardiovascular development, the single embryonic ventricle perfuses the blood to the right and left lateral mandibular aortic arches (AA1) through the bulbus arteriosus and the ventral aorta [22].…”

Section: Discussionmentioning

confidence: 99%

Analysis of early embryonic great-vessel microcirculation in zebrafish using high-speed confocal μPIV

et al. 2011

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In the developing cardiovascular system, hemodynamic vascular loading is critical for angiogenesis and cardiovascular adaptation. Normal zebrafish embryos with transgenically-labeled endothelial and red blood cells provide an excellent in vivo model for studying the fluid-flow induced vascular loading. To characterize the developmental hemodynamics of early embryonic great-vessel microcirculation in the zebrafish embryo, two complementary studies (experimental and numerical) are presented. Quantitative comparison of the wall shear stress (WSS) at the first aortic arch (AA1) of wild-type zebrafish embryos during two consecutive developmental stages is presented, using time-resolved confocal micro-particle image velocimetry (µPIV). Analysis showed that there was significant WSS difference between 32 and 48 h post-fertilization (hpf) wild-type embryos, which correlates with normal arch morphogenesis. The vascular distensibility of the arch wall at systole and the acceleration/deceleration rates of time-lapse phase-averaged streamwise blood flow curves were also analyzed. To estimate the influence of a novel intermittent red-blood cell (RBC) loading on the endothelium, a numerical two-phase, volume of fluid (VOF) flow model was further developed with realistic in vivo conditions. These studies showed that near-wall effects and cell clustering increased WSS augmentation at a minimum of 15% when the distance of RBC from arch vessel wall was less than 3 µm or when RBC cell-to-cell distance was less than 3 µm. When compared to a smooth wall, the WSS augmentation increased by a factor of ∼1.4 due to the roughness of the wall created by the endothelial cell profile. These results quantitatively highlight the contribution of individual RBC flow patterns on endothelial WSS in great-vessel microcirculation and will benefit the quantitative understanding of mechanotransduction in embryonic great vessel biology, including arteriovenous malformations (AVM). C.-Y. Chen et al. / Embryonic great vessel microcirculation vascular biology. Endothelial cells (EC) act as mechanosensors and translate the WSS loading into biochemical signals, which further control the downstream intracellular responses and modulate the gene/protein expression pathway [9,12,17]. The effects of altered hemodynamics are well established in the literature: the exposure of human aortic endothelial cells to an oscillatory flow (mean time-averaged shear stress 0.10 Pa) leads to an increase in cell proliferation, in contrast to the unidirectional laminar flow condition (mean shear stress > 1 Pa) [5]. Likewise, WSS loading plays a major role in the etiology of a number of cardiovascular diseases [40], such as atherosclerotic lesion development [18] and congenital heart defects. Particularly, the progression of high-flow arteriovenous malformations (AVM), which are associated with the mutations in the actin receptor-like kinase 1 (Alk1) gene, result in autosomal dominant vascular disease and hereditary hemorrhagic telangiectasia type 2 (HHT2) [3]. Recent studies us...

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Laser-scanning velocimetry: A confocal microscopy method for quantitative measurement of cardiovascular performance in zebrafish embryos and larvae

Cited by 59 publications

References 32 publications

Probing events with single molecule sensitivity in zebrafish and Drosophila embryos by fluorescence correlation spectroscopy

Probing events with single molecule sensitivity in zebrafish and Drosophila embryos by fluorescence correlation spectroscopy

The Cardiovascular Triad of Dysfunctional Angiogenesis

Analysis of early embryonic great-vessel microcirculation in zebrafish using high-speed confocal μPIV

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