title: Large-scale zebrafish embryonic heart dissection for transcriptional analysis.
abstract: 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.
authors: Max Delbrück Center for Molecular Medicine; Institute of Biochemistry and Biology, University of Potsdam; Institute of Molecular Biology, Medizinische Hochschule Hannover.
authors: Verónica A
authors: VA
authors: Lombardo
authors: Max Delbrück Center for Molecular Medicine; Institute of Biochemistry and Biology, University of Potsdam.
authors: Cécile
authors: C
authors: Otten
authors: Max Delbrück Center for Molecular Medicine; Institute of Biochemistry and Biology, University of Potsdam; Institute of Molecular Biology, Medizinische Hochschule Hannover; Salim.Seyfried@uni-potsdam.de.
authors: Salim
authors: S
authors: Abdelilah-Seyfried
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D006348:Cardiac Surgical Procedures / Q000379:methods / Q000662:veterinary*; D004210:Dissection / Q000379:methods / Q000662:veterinary; D020869:Gene Expression Profiling / Q000379:methods*; D006321:Heart / Q000196:embryology*; D009206:Myocardium / Q000737:chemistry*; D012333:RNA, Messenger / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction; D015027:Zebrafish / Q000196:embryology* / Q000601:surgery*; D029961:Zebrafish Proteins / Q000235:genetics
chemical_list: D012333:RNA, Messenger; D029961:Zebrafish Proteins
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