Chamber-specific ablation and reporter lines were generated using the standard I-SceI meganuclease transgenesis technique (details in Methods). To perform ventricular cardiomyocyte ablation, Tg(vmhc:mCherry-NTR) zebrafish were treated with 5 mM MTZ as previously described9 (link). For lineage tracing experiments, Tg(vmhc:mCherry-NTR;amhc:CreERT2;β-act2:RSG) zebrafish were treated with 10 µM 4-hydroxytamoxifen as previously described5 (link). For Notch inhibition studies, zebrafish were treated with 100 µM DAPT. Live imaging, heart contraction, immunofluorescence, and whole mount in situ hybridization were performed as described in Methods.
Protocol detail
Cardiomyocyte Ablation and Lineage Tracing in Zebrafish
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Act2
Cardiomyocyte
Dapt
Heart contraction
Hydroxytamoxifen
Immunofluorescence
In situ hybridization
Inhibition
Ventricular
Zebrafish
Corresponding Organization :
Other organizations : University of California, San Diego, University of California, Los Angeles, Sanford Burnham Prebys Medical Discovery Institute, Discovery Institute, University of California, San Francisco, Max Planck Institute for Heart and Lung Research
Top 5 similar protocols
Protocol cited in 48 other protocols
Variable analysis
independent variables
- Treatment with 5 mM MTZ
- Treatment with 10 µM 4-hydroxytamoxifen
- Treatment with 100 µM DAPT
- Ventricular cardiomyocyte ablation
- Lineage tracing
- Notch inhibition
- Live imaging
- Heart contraction
- Immunofluorescence
- Whole mount in situ hybridization
- Tg(vmhc:mCherry-NTR) zebrafish
- Tg(vmhc:mCherry-NTR;amhc:CreERT2;β-act2:RSG) zebrafish
- Not explicitly mentioned
- Not explicitly mentioned
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!