Artículo
Large-scale zebrafish embryonic heart dissection for transcriptional analysis
Fecha de publicación:
01/2015
Editorial:
Journal of Visualized Experiments
Revista:
Journal of Visualized Experiments
ISSN:
1940-087X
Idioma:
Inglés
Tipo de recurso:
Artículo publicado
Clasificación temática:
Resumen
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.
Palabras clave:
Developmental Biology
,
Dissection
,
Embryo
,
Heart
,
Issue 95
,
Rna
,
Rt-Qpcr
,
Zebrafish
Archivos asociados
Licencia
Identificadores
Colecciones
Articulos(CCT - ROSARIO)
Articulos de CTRO.CIENTIFICO TECNOL.CONICET - ROSARIO
Articulos de CTRO.CIENTIFICO TECNOL.CONICET - ROSARIO
Citación
Lombardo, Veronica Andrea; Otten, Cécile; Abdelilah Seyfried, Salim; Large-scale zebrafish embryonic heart dissection for transcriptional analysis; Journal of Visualized Experiments; Journal of Visualized Experiments; 95; 1-2015; 1-7
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