Award Date

5-1-2024

Degree Type

Dissertation

Degree Name

Doctor of Philosophy in Biological Sciences

Department

School of Life Sciences

First Committee Member

Kelly Tseng

Second Committee Member

Andrew Andres

Third Committee Member

Boo Shan Tseng

Fourth Committee Member

Mo Weng

Fifth Committee Member

Hui Zhang

Number of Pages

205

Abstract

The African Clawed Frog, Xenopus laevis, has been an indispensable model organism for over 100 years in the study of embryogenesis and development. In recent years, this vertebrate model has emerged as a robust tool for deciphering the cellular and molecular mechanisms underlying various modes of tissue, limb, and organ regeneration. In the context of eye repair, Xenopus offers a unique model, where the adult frog eye has limited regenerative capacity compared to its pre-metamorphic embryonic and tadpole forms, which exhibit a robust capacity for regrowth. Understanding the intricate molecular mechanisms that underlie this developmental shift in regrowth ability has the potential to inform our efforts to develop future directed regenerative therapies for eye regrowth. However, to reach this point, a crucial first step of identifying the participating cell populations and intrinsic regenerative strategies used by Xenopus must be decoded. To this end, we have developed a lineage tracing protocol with EosFP, enabling precise labeling of cells within Xenopus laevis throughout its developmental stages. This method, now adapted for use in studying Xenopus eye development, has broadened our knowledge of the internal dynamics of multipotent retinal stem cells within the developing eye, a task not yet achieved with previous labeling technologies. This approach allowed for the labeling of select cells within the eye field through embryonic development, revealing that retinal progenitor cells (RPCs) proliferate and differentiate within their region of origin in the eye field to form the subsequent mature anatomy of the retina. Furthermore, by photoconverting Eos protein in the eye field and subsequently removing the labeled optic vesicle, we have shown that RPCs within the post-surgical wound serve as a source of regenerative cells for the embryonic Xenopus eye. Surprisingly, our results also suggest that cells within the optic fissure of the regenerating eye might originate from a non-RPC lineage. This work advances our understanding of eye development and regeneration in Xenopus, while providing a powerful new tool for future research. By deciphering the natural regenerative strategies employed by Xenopus, we open the door for facilitating directed therapies that mirror proven natural regrowth processes.

Keywords

EosFP; Eye Development; Eye Field; Eye Regeneration; Eye Regrowth; Retinal Progenitor Cells

Disciplines

Biology | Cell Biology | Molecular Biology

File Format

pdf

File Size

3929 KB

Degree Grantor

University of Nevada, Las Vegas

Language

English

Rights

IN COPYRIGHT. For more information about this rights statement, please visit http://rightsstatements.org/vocab/InC/1.0/

Available for download on Thursday, May 15, 2025


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