Study On Expression And Biological Activity Of Human Neurturin Derived From Vero Cells

Neurotrophic factors are a kinds of peptides,which can promote neuron survival,growth,and differentiations.There are important actions in developping,differentiating,damaging and rapairing of nervous system.Neurturin (NTN) is a recently discovered neurotrophic factor that can act specifically on the dopaminergic neurons of the midbrain and motor neurons.This has important clinical significance for the treatment of nervous system diseases characterized by dopaminergic metabolic disorders,such as Parkinson's diseases.The recombinant NTN produced by methods of gene engineering can't go through the blood brain barrier and can be metabolized quickly then the T1/2 is short.It can bring about adverse effect if the recombinant NTN is given repeatedly.In view of this problem,gene therapy will be an good method for treating Parkinson's diseases,because it can be expressed constantly and act at target site.In the present study,we have done some researchs in constructing the eukarya expression vector of hNTN,transfecting the plasmid into Vero cells,acquiring the stability cells that express NTN constantly,assaying the activity of hNTN expressed by the stability cells.Firstly,the whole human NTN cDNA was cloned to plasmid pcDNA3.Secondly,the plasmid was transfected into Vero cells and made 80%these cells express NTN constantly.Lastly,the biological activities of NTN expressed by these cells were characterized.It shows that the production expressed by Vero cells stimulated growth of nerve fibers from the dorsal root ganglia of chick embryos and promoted survival of mesencephalon dopaminergic neurons of rat embryos in vitro.This is the first description of acquiring the vero cells that can express NTN constantly and protecting mesencephalon dopaminergic neurons.These results have provided a good foundation for development of NTN and treatment of Parksinson's diseases.