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  -