Efficient Down-Regulation of Glia Maturation Factor Expression in Mouse Brain and Spinal Cord

• Published in: Neurochemical research 2012-07, Vol.37 (7), p.1578-1583
• Main Authors: Zaheer, Smita, Wu, Yanghong, Yang, Xi, Thangavel, Ramasamy, Sahu, Shailendra K., Zaheer, Asgar
• Format: Article
• Language: English
• Subjects: Animals; Base Sequence; Biochemistry; Biomedical and Life Sciences; Biomedicine; Brain; Brain - metabolism; Cell Biology; Central nervous system; Down-Regulation; Enzyme-Linked Immunosorbent Assay; Experimental allergic encephalomyelitis; Expression vectors; Glia maturation factor; Glia Maturation Factor - metabolism; Inflammation; Mice; Mice, Inbred C57BL; Neurochemistry; Neurology; Neurosciences; Original Paper; RNA; RNA, Small Interfering; Spinal cord; Spinal Cord - metabolism; Thorax
• ISSN: 0364-3190; 1573-6903
• DOI: 10.1007/s11064-012-0753-x

Long-lasting siRNA-based down-regulation of gene of interest can be achieved by lentiviral-based expression vectors driving the production of short hairpin RNA (shRNA). We investigated an attractive therapeutic approach to target the expression of proinflammatory GMF by using lentiviral vector encoding GMF–specific shRNA to reduce GMF levels in the spinal cord and brain of mice. To determine the effect of GMF–shRNA on GMF protein levels, we performed quantitative ELISA analysis in brain and in thoracic, cervical and lumbar regions of spinal cord from mice followed by GMF–shRNA (G-shRNA) or control shRNA (C-shRNA) treatments. Our results show a marked reduction of GMF protein levels in brain and spinal cord of mice treated with GMF–shRNA compared to control shRNA treatment. Consistent with the GMF protein analysis, the immunohistochemical examination of the spinal cord sections of EAE mice treated with GMF–shRNA showed significantly reduced GMF-immunoreactivity. Thus, the down-regulation of GMF by GMF–shRNA was efficient and wide spread in CNS as evident by the significantly reduced levels of GMF protein in the brain and spinal cord of mice.