Regeneration in Zebrafish
Focus:
I will be using CREATE in BIO 410 Molecular Cell Biology. This is a senior level elective that includes a three hour/week lab.
The rationales for choosing the topic of regeneration in zebrafish:
1) students express great interest in the topic of regeneration.
2) zebrafish are available for the performing selected methods from the papers in the associated lab.
This class has a lab that meets for three hours/week so my goal will be to incorporate some of the methods we learn and discuss from the articles into an applied learning experience. In past semesters, I have allowed students to design and carry out their own mini research projects in this lab - applying a grant panel activity before mid semester would allow students more thorough feedback before they actual embark on their projects.
Overview:
Applicable for Courses:
Molecular Cell BiologyEducational Level:
Upper-levelRoadmap Objectives:
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- Article: Heart regeneration in zebrafish. (2002) Poss et al. Science 298:2188-2190.
- Content area/major concepts: stem cells, regeneration, cell and tissue types, cell cycle, chromatin structure and function, temperature sensitive mutations.
- Methods or technology used to obtain data: Care and handling of zebrafish, ventricular resection, histology, in situ hybridization, BrdU labeling, Immunofluorescence,
- How the CREATE strategy was used:
- Biggest teaching challenge:
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- Article: Retinoic acid production by endocardium and epicardium is an injury response essential for zebrafish heart regeneration. (2011) Kikuchi et al. Dev. Cell 20: 397-404.
- Content area/major concepts: zebrafish development, signal transduction, cell differentiation, apoptosis, inflammation?
- Methods or technology used to obtain data: zebrafish embryo collection and handling, transmission electron microscopy, TUNEL assays, Construction of transgenic animals, Isolation of RNA, RT-PCR, flow cytometry, heart explant culture
- How the CREATE strategy was used:
- Biggest teaching challenge:
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- Article: The regenerative capacity of zebrafish reverses cardiac failure caused by genetic cardiomyoctye depletion. (2011) Wang et al. Development 138:3421-3430
- Content area/major concepts: cardiac physiology and electrocardiograms
- Methods or technology used to obtain data: genetic cell ablation, heat stress, physiological experiments (swim tunnel respirometer), histology, Cre-mediated recombination
- How the CREATE strategy was used:
- Biggest teaching challenge:
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- Article: Clonally dominant cardiomyocytes direct heart morphogenesis. (2012) Gupta and Poss. Nature 484: 479-485.
- Content area/major concepts: organ development, morphogenesis, fate mapping,
- Methods or technology used to obtain data: Continuation of Cre/lox recombination and transgenesis, “Brainbow” transgenesis
- How the CREATE strategy was used:
- Biggest teaching challenge: