Adult teleost fish and urodele amphibians can regenerate entire amputated appendages. By contrast, regenerative healing of adult mammalian limbs is limited to the very tips of digits. One of the key challenges in developmental biology is to understand how and why tissue regeneration occurs. The hallmark of limb or fin regeneration is formation of a blastema, a mesenchymal structure that contains progenitor cells for new skeletal elements. As regeneration proceeds, blastemal cell proliferation and patterning are regulated such that lost tissues of correct size and shape are replaced, a phenomenon called positional memory. Skeletal osteoblasts in the blastema and, ultimately, the regenerated fin rays, derive from the proliferation of pre- existing osteoblasts, while an alternative source(s exists if the primary osteoblast source is disabled. Activating and inhibitory factors influence regenerative growth, and region-specific maintenance of patterning transcription factors in fin cells throughout life is a component of positional memory. Despite recent advances in our understanding of appendage regeneration, there remain key deficiencies in the field. First, there have been no published attempts to image and quantify the in vivo dynamics of cell proliferation during appendage regeneration, analyses that would illuminate how complex pattern is restored after injury. Second, a limited set of traditional developmental signaling factors has been assessed for function during regeneration. The overall goal of this proposal is to define mechanisms that regulate cell proliferation as size and pattern are restored to an amputated appendage. 1) We will apply new technologies to monitor in vivo cell cycle progression and growth indicators in transgenic zebrafish, to create a spatiotemporal map of cell proliferation during fin regeneration. 2) We will use new reagents to define whether and how signaling by the secreted factor Leptin influences zebrafish fin regeneration. 3) We will define mechanisms by which two genes implicated from a mutagenesis screen, the extracellular matrix component laminin beta 1a and the voltage-gated calcium channel calcium homeostasis modulator 1, influence fin regeneration. With these approaches, we will test the hypothesis that blastemal cell proliferation is controlled by a complex microenvironment of activating and inhibitory influences. This work will increase understanding of developmental regulation during vertebrate tissue regeneration, and provide important perspective for comprehending, and perhaps changing, the existing limitations in regenerative capacity of human tissues.

Public Health Relevance

This work will increase understanding of developmental regulation during vertebrate tissue regeneration, and provide important perspective for comprehending, and perhaps changing, the existing limitations in regenerative capacity of human tissues.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM074057-09
Application #
8770139
Study Section
Development - 1 Study Section (DEV1)
Program Officer
Haynes, Susan R
Project Start
2006-05-01
Project End
2018-06-30
Budget Start
2014-07-10
Budget End
2015-06-30
Support Year
9
Fiscal Year
2014
Total Cost
$321,850
Indirect Cost
$116,850
Name
Duke University
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
044387793
City
Durham
State
NC
Country
United States
Zip Code
27705
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Han, Yanchao; Poss, Kenneth D (2016) Accessories to Limb Regeneration. Dev Cell 37:297-8
Nichols, James T; Blanco-Sánchez, Bernardo; Brooks, Elliott P et al. (2016) Ligament versus bone cell identity in the zebrafish hyoid skeleton is regulated by mef2ca. Development 143:4430-4440
Tornini, Valerie A; Puliafito, Alberto; Slota, Leslie A et al. (2016) Live Monitoring of Blastemal Cell Contributions during Appendage Regeneration. Curr Biol 26:2981-2991
Kang, Junsu; Hu, Jianxin; Karra, Ravi et al. (2016) Modulation of tissue repair by regeneration enhancer elements. Nature 532:201-6
Chen, Chen-Hui; Merriman, Alexander F; Savage, Jeremiah et al. (2015) Transient laminin beta 1a Induction Defines the Wound Epidermis during Zebrafish Fin Regeneration. PLoS Genet 11:e1005437
Kang, Junsu; Karra, Ravi; Poss, Kenneth D (2015) Back in Black. Dev Cell 33:623-4
Tornini, Valerie A; Poss, Kenneth D (2014) Keeping at arm's length during regeneration. Dev Cell 29:139-45
Gemberling, Matthew; Bailey, Travis J; Hyde, David R et al. (2013) The zebrafish as a model for complex tissue regeneration. Trends Genet 29:611-20

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