Date Thesis Awarded

5-2016

Document Type

Honors Thesis

Degree Name

Bachelors of Science (BS)

Department

Neuroscience

Advisor

Margaret Saha

Committee Member

Peter Kemper

Committee Member

Diane Shakes

Committee Member

Gregory Smith

Abstract

Spontaneous intracellular calcium activity has been implicated in a host of processes related to nervous system development, including neural induction, neural tube closure, and synaptogenesis. One of these calcium-influenced processes, neurotransmitter phenotype specification, involves the acquisition of the correct balance and patterning of excitatory and inhibitory neurons, and its regulation is vital to proper nervous system functionality. While a high frequency of intracellular calcium transients in presumptive neurons during development has been correlated with an inhibitory fate, the persistence of this phenomenon in in vitro models has not been conclusively demonstrated. Additionally, we believe that current methods of calcium activity analysis, which are limited to counting fluorescent indicator spikes above a particular threshold, is limiting. To this end, we employed Xenopus laevis presumptive neural tissue as an in vitro model system, imaging calcium activity in developing neural cells. This data was analyzed via a novel pipeline that uses fluorescence trace entropy as a comparative metric rather than relying on predetermined parameters to define particular features (i.e., spikes or waves). Use of this analysis method revealed differences in calcium activity across development, with cells dissected from younger embryos displaying more entropic calcium activity that gradually decreased across development. Relatively small differences were found between cells positive for the expression of particular neural marker genes and cells that did not express these genes, and these differences varied across developmental time points. Most notably, cells positive for different specific neural marker genes displayed significantly different levels of calcium activity entropy from one another. As a whole, these results provide support for the hypothesis that particular patterns of calcium dynamics are associated with the expression of particular genes involved in the neuronal differentiation process.

Creative Commons License

Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 License.

Available for download on Monday, May 13, 2019

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