Characterizing Baz1a: Designing Cell Labeling Strategy and Development of Hi-Res Neuron Model

Overview

The human brain and the neurons that compose it remain some of the greatest mysteries facing scientists today. Though the basic structure of a neuron is known, an updated and detailed model of a single neuron is an excellent first step in the quest to understand how neurons interact, and on a higher level, how the brain works. By combining multiple modern neuron imaging techniques and focusing them on just one type of neuron, enough data can be gathered to contribute to a high-resolution model of a neuron. Within my lab, the eventual goal was to map the inhibitory synapses of the neuron of interest, creating a detailed in-vivo synaptic mapping of the neuron. Following the selection of the Baz1a neuron, an excitatory cell expressed in Layer 2/3 of the visual cortex, as the neuron of interest, work began to design a method for labeling Baz1a and imaging it in-vivo. A plasmid was constructed to include a Baz1a promoter and a Yellow Fluorescent Protein, then was transfected in cultured neurons to confirm cell expression. When comparing Baz1a-YFP images with the control ubiquitin-BFP images, it was suggested that Baz1a may not be as selective a promoter as it was hoped to be. This selectivity issue was further investigated in mice brains, where out of 100 cells, it was found that 23% of cells were only labeled by the ubiquitin promoter, 2% were labeled only by the Baz1a promoter, and 75% were doubly expressed. Because roughly 75% of the cells in this tissue were being labeled by both the Baz1a promoter and a ubiquitin promoter, it was suggested that the Baz1a promoter was not as selective as previously thought. In order to confirm this issue of Baz1a selectivity, the Baz1a-YFP brains must undergo fluorescent in-situ hybridization, which would detect the location of Baz1a neurons and give insight to which neurons are being correctly versus incorrectly labeled by the Baz1a promoter.

Competition history

• AJAS 2025 Category not listed

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