부산대학교 도서관

The objective of this work was the generation of an animal model of the SCA2 disease for future studies on the benefits of therapeutic molecules and the underlying neuropathological mechanisms in this human disorder. The transgenic fragment was microinjected into pronuclei of B6D2F1 X OF1 mouse hybrid strain. For Northern blots, RNAs were hybridized with a human cDNA fragment from the SCA2 gene and a mouse b-actin cDNA fragment. Monoclonal antibodies directed at the N-terminal of the ataxin 2 protein with 22Q were used for Western blot analysis. A rotating rod apparatus was utilized to measure motor coordination of mice. Immunohistochemical detection of Purkinje neurons was performed with anti-calbindin 28K as the primary antibody. An ubiquitous expression of the SCA2 transgene with 75 CAG repeats regulated by the SCA2 self promoter was obtained after the generation of our transgenic mice. The analysis of transgenic mice revealed significant differences of motor coordination compared with the wild type littermates. A specific degeneration of Purkinje neurons and transgene over-expression in the brain, liver and skeletal muscle, rather than in lungs and kidneys was also observed, resembling the expression pattern of the ataxin 2 in humans. [ABSTRACT FROM AUTHOR]