LE3 .A278 2015

2015

Smith, Todd
Bulman, Dennis

Acadia University

Bachelor of Science

Honours

Biology

Biology

Myoclonus dystonia (MD) is a rare neuromuscular genetic disorder characterized by uncontrolled movements of the limbs and painful muscle spasms (dystonia). The disease occurs in approximately 1 in 200,000 individuals. The clinical onset of MD generally occurs within childhood, and the majority of cases are diagnosed during the first two decades of life. Mutations in SGCE, the gene that codes for epsilon sarcoglycan, have been linked to roughly 40% of MD cases. SGCE codes for a transmembrane protein and has high levels of expression in the cerebellum. Another potential MD gene candidate is CD108131; this gene does not have an open reading frame and codes for a long non-coding RNA. Mutations in this gene possibly inhibit or hinder its ability to regulate the expression of other genes or proteins linked to this disease. Specifically, CD108131 may directly or indirectly regulate the expression of SGCE at a transcriptional or translational level. This project focuses on using this model to link CD108131 to MD. To do so, mutated and wild type CD108131 was amplified from genomic DNA isolated from patient and control individuals. The resulting inserts were cloned into a mammalian vector and transfected into HEK-293 cells. A western blot of the epsilon sarcoglycan protein was performed on harvested cellular protein. Overexpression of wild type CD108131 increased expression of SGCE to twice that of the empty control, whereas overexpression of mutant CD108131 only resulted in a 27% increase. This confirmed the hypothesis that CD108131 is linked to the expression of SGCE, and that mutations in this gene obstruct its ability to regulate expression.

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https://scholar.acadiau.ca/islandora/object/theses:1255