Date Thesis Awarded

5-2023

Access Type

Honors Thesis -- Access Restricted On-Campus Only

Degree Name

Bachelors of Science (BS)

Department

Biology

Advisor

Margaret S. Saha

Committee Members

Daniel A. Cristol

Shantá D. Hinton

Noah M. Lemos

Abstract

Calcium is a ubiquitous, highly conserved universal messenger that mediates an array of functions such as neuronal and neuromuscular synaptic transmission, the stress response, wound healing, cardiac myocyte function, brain homeostasis maintenance, cell proliferation, and the immune response in mature organisms; this highly regular, stereotypical calcium activity in mature organisms has been extensively studied and is well understood (Vig et al., 2009; Fearnley et al., 2011; Patergnani et al., 2020). Calcium activity has also been implicated as a key regulator during the early development of organisms in processes such as neural induction, neuronal fate determination and differentiation, and organogenesis (Paudel et al., 2018). However, despite its universal presence in developing embryos, the spontaneous calcium activity that exists in embryonic systems is less well-characterized. This results from a dearth of studies, lack of consensus on analytical techniques, and contradictory results from literature in this field. Past studies on calcium activity during early embryonic development are also limited in that they are largely correlational rather than causal or functional.

This thesis uses several different approaches to explore the role of calcium activity in early embryonic development. Chapter 1 provides a comprehensive review of the tweety gene family—which encodes calcium responsive gated chloride channels—by describing its structure, evolution, expression during development and adulthood, biochemical and cellular functions, and role in human disease. Then, chapter 2 discusses analysis of time-series data collected on embryos that were microinjected with GCaMP mRNA (a genetically-encoded calcium indicator that enables fluorescent visualization of calcium activity) and imaged for calcium activity at cellular resolution over a one hour period using fluorescent confocal microscopy. Importantly, this time-series analysis reveals that there is a lack of standardization in the methodology used to analyze time-series data of embryonic calcium activity imaging. This insight inspired the investigation described in chapter 3, which is a comprehensive review and critique of the existing methodologies used to detect and analyze imaging data of embryonic calcium activity. The review serves as a call to action to standardize the techniques used to analyze imaging data of embryonic calcium activity. Finally, chapter 4 experimentally examines the effects of calcium activity on early embryonic development by perturbing embryonic calcium dynamics using an automated calcium perturbation device that I invented. Ultimately, elucidating the role of calcium activity on early embryonic development through this study helps create the molecular framework necessary for the development of promising therapeutic strategies that target developmental diseases and disorders implicated by dysregulated embryonic calcium dynamics.

Available for download on Saturday, May 16, 2026

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