Date Thesis Awarded

4-2014

Document Type

Honors Thesis

Degree Name

Bachelors of Science (BS)

Department

Biology

Advisor

Daniel Cristol

Committee Member

Mark Forsyth

Committee Member

Elizabeth Harbron

Abstract

Stem cell specification, maintenance, and differentiation are imperative to proper organogenesis and tissue maintenance. Understanding the mechanisms and steps of these developmental processes is invaluable to advancing tissue grafting, cancer treatment, and other organ therapies. The Drosophila melanogaster testis is one of the best-characterized systems for studying these processes. Currently, studies in the Drosophila testis have thoroughly characterized when stem cells develop and what the resulting adult testis morphology will be. A major question that remains to be answered is how stem cells develop – what controls what cells become functional stem cells and how stem cell fates are dictated. To seek answers to this question, I propose a focus on live-cell imaging studies of the developing Drosophila testis. In order to perform live-cell imaging studies in this developmental system, it is necessary to successfully design protocols for: (1) gonad isolation, (2) testis culture, (3) imaging of developing gonads over time in three‑dimensions (3D), and (4) tracking cell movement in developing gonads over time. In this thesis, I provide evidence for successful completion of the first three steps towards live-cell imaging, as well as preliminary studies of cell tracking. Testes at different key stages of development, (1) from gonad coalescence through germline stem cell (GSC) niche formation and (2) from GSC niche formation to spermatogonial differentiation, were studied for evidence of successful live-cell imaging. These key developmental events were chosen because they are characterized by major morphological shifts that can be clearly observed via live-cell imaging. These findings provide a promising start to a line of research inquiry that could answer many key questions that have remained elusive to investigators focused on understanding stem cell development (Slack, 2008).

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