The Role Of Stil Gene In The Proliferation Of PC12Cell And Retina Cell In Zebrafish And The Establishment Of A Zebrafish Model Of Parkinson’s Disease

Cell proliferation is widespread in nature, from the lower animal and plant to higher organism, they all will die without the cell proliferation, and cell proliferation during the growth of the organism is very rigorous and precise control, once the part of the regulation network function change, it will disrupted the cell proliferation, and affect the normal growth and development of the organism or the realization of the physiological function.Cell proliferation is indispensable to the regeneration of tissue and organ, the regeneration and repair of tissue and organ depends on proliferation, when a part of tissue or organ damage, it will start the corresponding signaling pathway to regulate the repair of tissue and organ in many species in nature, lower organisms showed stronger regeneration ability than higher biological, such as polyps, planaria,even African clawed frog fish such as salamanders, while as a higher biological--mammals during the long evolution have lost the ability to regenerate.We use zebrafish as a model because it has strong regeneration ability, and its genome is very high homology with the human genome, and it has important research significance very much.Stil gene, which originally was found from T cells acute lymphocytic leukemia, with the deepening of the research, scientist also found that when Stil gene mutate, it will lead to microcephaly disease in humans, and it also regulate the development of bilateral symmetry through Shh signaling pathways.besides, researchers found that the expression of Stil gene in many cancer cells is higher, further study showed the product of Stil gene is a part of centrosome, and it plays an improtant role in the assembly and copy of centrosome, when Stil gene is knock down, cells lack the centrosome; When Stil gene expressed too much, it appeared two or even more centrosome in the cell.First, We use PC12nerve cells as the research model, and we found STIL proteins in PC12cells not only existed in the cytoplasm, also in the nucleus, and it suggests that STIL genes may have other unknown functions; We also confirmed the STIL protein in PC12cells can also interact with the SUFU and GLI1, but also there may be a heterotrimeric protein complex; When Stil gene is Stilent, cell proliferation slow down, and the marker gene of hh signaling pathway gene--glil,its expression also decreased; when Stil gene is overexpressed, PC12cell proliferation speed is accelerated, the expression of glil is raised, the expression of target genes cyclinD2also increased, so we speculated that Stil gene regulate the proliferation through cyclinD2to affect the activity of CDK, finally affecting the cell cycle.Besides, We utilized the zebrafish mutant (Stil) and wild type zebrafish to study, and found the proliferation of dopamine cells in retinas show different rate after injection of6-OHDA neurotoxin in retinas. Dopamine cells proliferation in nbb+/-is slower than wild type, and it’s only about half of the wild type, and when we knock down sufu gene with MO-sufu, glil gene is activated, the proliferation of dopamine cells speed up, and during the process of spontaneous repair after injury, Stil gene and glil genes are involved in the process of cell proliferation in the zebrafish retina, in short, Stil can promote dopamine cells proliferation.Then, we took the zebrafish as an object, based on mature experiment technology to inject6-OHDA to its intraocular to kill dopamine cells specifically, and then by means of the identification of behavioural and physiological,we found its characteristics are in line with the characteristics of Parkinson’s patients, so we set up a zebrafish model of Parkinson’s disease. This model construction is simple and rapid, so it can be used as high throughput screening model for Parkinson’s disease drug.