Abstract

The first dynein mutation found in humans that caused disease was a cytoplasmic dynein 1 heavy chain (DYNC1H1 in humans) p.His306Arg mutation, first described by Weedon et al. in 2011. This mutation caused Charcot-Marie-Tooth (CMT) subtype 2O. CMT has a prevalence of approximately 1 in 2500 people, making it the most common hereditary neuromuscular disorder. Cytoplasmic dynein 1 is used by eukaryotic cells for minus-end directed microtubule-based transport of cargo. One such cargo is Rab7, a late endosomal marker. The purpose of this study is to identify the effects of this mutation on the transport of GFP-tagged Rab7 cargo in neurons from wild type (HH), heterozygous mutant (HR), and homozygous mutant (RR) mice harboring a DYNC1HI His306Arg mutation. Mouse embryos were euthanized, dissected to collect the hippocampal and cortical brain tissues, and these tissues were digested to isolate neurons. Nucleofection was used to introduce the exogenous GFP-Rab7 gene construct. These neurons were plated and imaged at 10 days in vitro using wide-field epifluorescence microscopy to generate image stacks of fluorescent GFP-Rab7 vesicles. Kymograph analysis was performed on the image stacks using MetaMorph software to measure several characteristics of movement. Statistical analysis of the data from each of the three genotypes shows there is no significant difference in Rab-7 transport between the three genotypes.

Thesis Completion

2017

Semester

Spring

Thesis Chair/Advisor

King, Stephen

Degree

Bachelor of Science (B.S.)

College

College of Medicine

Department

Burnett School of Biomedical Sciences

Degree Program

Biomedical Sciences

Location

Orlando (Main) Campus

Language

English

Access Status

Open Access

Release Date

11-1-2017

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Cell Biology Commons

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