Date of Graduation

5-2023

Document Type

Thesis

Degree Name

Bachelor of Science in Biology

Degree Level

Undergraduate

Department

Biological Sciences

Advisor/Mentor

Paré, Adam

Committee Member/Reader

Dowling, Ashley

Committee Member/Second Reader

Beaulieu, Jeremy

Committee Member/Third Reader

Chapman, Kate M.

Abstract

Embryonic development is a complex process requiring dynamic cell movements to create complex tissue structures. We assume that these embryonic dynamic remodeling events are highly energy intensive and that this energy expenditure is mediated by mitochondria. However, this has been difficult to demonstrate in live embryos. Convergent extension of the Drosophila neuroectoderm is a promising system to study the bioenergetics of tissue remodeling because it changes from a static to a dynamically remodeling tissue with temporal precision. We hypothesize that the switch from static to dynamic development at the onset of convergent extension will be accompanied by an increase in metabolic energy production. Our method of measuring metabolic production is through the use of the Seahorse XF HS Mini Analyzer, which is capable of measuring bioenergetic processes in small numbers of cells. Notably, the Seahorse Analyzer measures Oxygen Consumption Rate (OCR) and Extracellular Acidification Rate (ECAR), which are indicators of energy production. I expect that an increase in metabolic production will be indicated by an increase in OCR and a drop in ECAR. By measuring these bioenergetic markers before and during convergent extension, the effects of dynamic tissue development on metabolism can be uncovered. All empty control wells resulted in OCR and ECAR readings near zero, and all wells with embryos measured between 100-300 pmol/min OCR. The Seahorse Analyzer could detect the presence of live embryos but was unable to detect consistent differences between one and multiple embryos in a well. These results were incapable of demonstrating whether metabolism increases or remains constant, as we did see an effect, but the data lacked consistency and the effect did not scale with the number of embryos.

Keywords

Embryogenesis; Developmental Biology; Mitochondrial Dynamics; Drosophila; Tissue Remodeling; Convergent Extension

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