Graduation Year
2024
Document Type
Master's Thesis
Degree
Master of Science
Program
Biological Science
Program Director
Meredith Protas, PhD
First Reader
Danielle Tan
Second Reader
Manuel Lopez
Abstract
This study focused on developing a set of advanced histological tools to conduct spatial analysis of the central nervous system (CNS), improving upon traditional methods, and addressing their shortcomings. We applied these methods to explore CNS drug biodistribution. With the goal of defining regions of tissue in which the drug is distributed and the specific cell types that uptake it, we employed tissue clearing, immunofluorescent staining of unique cellular population-identifying antigens, and light-sheet fluorescence microscopy (LSFM) techniques to image adult mouse hemispheres in a variety of conditions and compared these techniques to traditional immunofluorescence (IF) on thin tissue sections. In addition, we used high-content spatial proteomics to observe the regional and cellular subtype localization and concentration-dependent changes to a reporter-tagged adeno-associated virus (AAV) following intracerebroventricular (ICV) delivery. Here we have improved upon standard technical counterparts and used advanced histological techniques, 3D clearing combined with LSFM and high-content spatial proteomics, to shed light on regional and cell-specific drug biodistribution within the brain.
Included in
Animal Experimentation and Research Commons, Cell Biology Commons, Developmental Neuroscience Commons, Molecular and Cellular Neuroscience Commons, Nervous System Commons, Neurosciences Commons, Other Chemicals and Drugs Commons, Other Neuroscience and Neurobiology Commons, Therapeutics Commons, Tissues Commons