IGF-1 stimulation via GSK-3β and screening for genes conferring protection of SHEP-1 cells against MPP+ toxicity

• Main Author: Li, Jikun
• Other Authors: Feng Zhiwei
• Format: Final Year Project
• Language: English
• Published: 2009
• Subjects: DRNTU::Science::Biological sciences
• Online Access: http://hdl.handle.net/10356/19313
• Institution: Nanyang Technological University

1-methyl-4-phenylpyridinium (MPP+) causes Parkinsonism and has been widely used in experimental models of Parkinson’s disease. MPP+ was used to induce apoptosis in human dopaminergic neural SHEP-1 cells in order to identify and characterise genes that may protect against MPP+ toxicity. In this study, we had demonstrated that BIO, a glycogen synthase kinase 3β (GSK-3β) inhibitor, is able to protect SHEP-1 cells against MPP+ toxicity. With these results, we can now focus on the Akt1/GSK-3β signalling pathway following our observation that Insulin-like Growth Factor 1 (IGF-1) confers SHEP-1 cells protection against MPP+ toxicity in a dose-dependent manner. In an effort to identify more genes that may play a role in protection against MPP+ toxicity, we used random insertion of an enhanced retroviral mutagen (ERM) cassette in SHEP-1 cells followed by 3’ Rapid Amplification of cDNA (3’-RACE) to select for clones resistant to MPP+ toxicity . This led to the identity of CARHSP1, MRP63, and PFN2 as potential genes which might be associated with MPP+ induced apoptosis. Early diagnosis of Parkinson’s disease coupled with early administration of IGF-1 could help prevent further death of dopaminergic neurons slowing the progression of the disease. It is also possible that drugs inhibiting GSK3-β can be administered in the SNpc to achieve similar effects.