Jasmine Zeki

Date of Award

2012

Document Type

Thesis

Degree Name

Bachelors

Department

Natural Sciences

First Advisor

Beulig, Alfred

Keywords

Neurobiology, Mice Visual Cortex, Two-Photon Imaging

Area of Concentration

Neurobiology

Abstract

This thesis examines new procedures used to study the functional and structural properties in the primary visual cortex (V1) in mice. Chronic window surgery was used to create an opening in the mice skulls that can easily be continually accessed and imaged. Two-Photon microscopy was used to achieve a deeper view into a live mouse than would be available in other forms of microscopy. Intrinsic Imaging functioned to establish the area viewed under the Two-Photon microscope as V1 by showing blood flow with two orientations of a moving bar stimulus. Calcium Imaging was used to show cell activity when a mouse is presented with two different moving bar stimuli which have varying moving bar orientations. Cells were found to be tuned or not and then compared to other cells in the same area to examine the effect of proximity of cells on creating functional relationships between them. Single cell electroporation was used to introduce a rabies plasmid to a single cell while keeping it alive. Once a successful electroporation occurred, the cell received an injection of a modified rabies virus. If this step was successful, the pre-synaptic connections to a single post-synaptic cell would be demonstrated. Overall, the results showed that the procedures used to obtain data were very successful. Intrinsic Imaging showed blood flow orthogonal to each other when presented with two different orientations of moving bars. Calcium Imaging showed changing results from groups of cells that were similarly tuned to areas of cells that were all differently tuned. This raised the question of the effect of the two different moving bar stimuli on the results obtained. Single cell electroporation proved to be a difficult but good method of giving a single cell the plasmid required for later rabies infection. The rabies infection was successful for those cells which were successfully electroporated. Slices of the successfully infected brains were mounted on microscope slides so that pre-synaptic cells could be counted. In conclusion, the methods used in this study were a successful way of gathering functional and structural data of mice visual cortex separately. The next step is to obtain method to connect these two sets of data to find a connection between functional and structural data.

Rights

This bibliographic record is available under the Creative Commons CC0 public domain dedication. The New College of Florida, as creator of this bibliographic record, has waived all rights to it worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law.

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