Year

2020

Season

Summer

Paper Type

Master's Thesis

College

College of Arts and Sciences

Degree Name

Master of Science in Biology (MS)

Department

Biology

NACO controlled Corporate Body

University of North Florida. Department of Biology

First Advisor

Dr. Judith Ochrietor

Second Advisor

Dr. Fatima Rehman

Rights Statement

http://rightsstatements.org/vocab/InC/1.0/

Third Advisor

Dr. Beth Stotz-Potter

Abstract

Chronic inflammation is a hallmark of many neurodegenerative disorders. Although the central nervous system (CNS) can stave peripheral pathogens from crossing the blood-brain barrier (BBB) through a network of continuous endothelia, astrocytes, and pericytes, prolonged exposure to a pathogen can comprise this barrier. Basigin, a cell adhesion molecule, is found on the surface of endothelial cells and has been demonstrated to interact with toll-like receptor 4 (TLR4). TLR4 recognizes lipopolysaccharide (LPS), found on the outer membrane of Gram-negative bacteria. The activation of TLR4 produces pro-inflammatory cytokines, like IL-6. The present study aims to address the expression pattern of Basigin gene products and TLR4 in brain and retinal tissue stimulated with LPS for a variation of time to mirror acute and chronic inflammation, as well as different life stages to determine whether the expression pattern is dynamic. Isolated brain tissue and neural retina from mice at postnatal day 7, 30 and 180 were incubated in DMEM ± LPS for 3, 6, 12, or 24 hrs. Total RNA and protein were purified from the isolated tissue and used in quantitative reverse transcription PCR (qRT-PCR) and direct enzyme-linked immunosorbent assay (ELISA). Basigin, TLR4, and IL-6 were localized in brain tissue via immunohistochemistry. The results of the study suggest Basigin is highly expressed on microvasculature endothelial cells. The expression pattern of Basigin was not only dependent on length of exposure to LPS, but also age. Basigin’s differential expression in 7- and 180-, but not 30-day old animals suggest pathogenic influence is more likely in neonatal and adult, but not adolescent mice. The pattern of TLR4 expression did not mirror that of Basigin gene products, indicating Basigin’s role may not be to associate with TLR4, but may associate with other pro-inflammatory proteins, or may be acting in its role as an inducer of matrix metalloproteinases.

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