TY  - JOUR
A1  - Lombardo, Veronica A.
A1  - Otten, Cecile
A1  - Abdelilah-Seyfried, Salim
T1  - Large-scale Zebrafish Embryonic Heart Dissection for Transcriptional Analysis
JF  - Journal of visualized experiments
N2  - The zebrafish embryonic heart is composed of only a few hundred cells, representing only a small fraction of the entire embryo. Therefore, to prevent the cardiac transcriptome from being masked by the global embryonic transcriptome, it is necessary to collect sufficient numbers of hearts for further analyses. Furthermore, as zebrafish cardiac development proceeds rapidly, heart collection and RNA extraction methods need to be quick in order to ensure homogeneity of the samples. Here, we present a rapid manual dissection protocol for collecting functional/beating hearts from zebrafish embryos. This is an essential prerequisite for subsequent cardiac-specific RNA extraction to determine cardiac-specific gene expression levels by transcriptome analyses, such as quantitative real-time polymerase chain reaction (RT-qPCR). The method is based on differential adhesive properties of the zebrafish embryonic heart compared with other tissues; this allows for the rapid physical separation of cardiac from extracardiac tissue by a combination of fluidic shear force disruption, stepwise filtration and manual collection of transgenic fluorescently labeled hearts.
KW  - Developmental Biology
KW  - Issue 95
KW  - zebrafish
KW  - embryo
KW  - heart
KW  - dissection
KW  - RNA
KW  - RT-qPCR
Y1  - 2015
U6  - https://doi.org/10.3791/52087
SN  - 1940-087X
IS  - 95
PB  - JoVE
CY  - Cambridge
ER  - 
TY  - JOUR
A1  - Dietrich, Ann-Christin
A1  - Lombardo, Veronica A.
A1  - Abdelilah-Seyfried, Salim
T1  - Blood flow and Bmp signaling control endocardial chamber morphogenesis
JF  - Developmental cell
N2  - During heart development, the onset of heartbeat and blood flow coincides with a ballooning of the cardiac chambers. Here, we have used the zebrafish as a vertebrate model to characterize chamber ballooning morphogenesis of the endocardium, a specialized population of endothelial cells that line the interior of the heart. By combining functional manipulations, fate mapping studies, and high-resolution imaging, we show that endocardial growth occurs without an influx of external cells. Instead, endocardial cell proliferation is regulated, both by blood flow and by Bmp signaling, in a manner independent of vascular endothelial growth factor (VEGF) signaling. Similar to myocardial cells, endocardial cells obtain distinct chamber-specific and inner- versus outer-curvature-specific surface area sizes. We find that the hemodynamic-sensitive transcription factor Klf2a is involved in regulating endocardial cell morphology. These findings establish the endocardium as the flow-sensitive tissue in the heart with a key role in adapting chamber growth in response to the mechanical stimulus of blood flow.
Y1  - 2014
U6  - https://doi.org/10.1016/j.devcel.2014.06.020
SN  - 1534-5807
SN  - 1878-1551
VL  - 30
IS  - 4
SP  - 367
EP  - 377
PB  - Cell Press
CY  - Cambridge
ER  -