Microglia And Neural Stem Cell Co-transplants With Over Expression Of Nurr1 Improves Functional Outcome In Parkinsonian Rats

ObjectiveTo explore the improved effects of over expression of Nurrl in microglia and neural stem cell co-transplants on PD rats with impaired motor function.Methods1.Neural stem cells were isolated from the Ventral mesencephalic neural zone of fetal SD rats atembryonic day 14(E14)under sterile conditions;Microglia were isolated from neonatal SD rats.After purification and identification,the Microglia and Neural stem cells were over expression of Nurrl.2.Stereotaxic injections of 6-hydroxydopamine(6-OHDA)were performed SD male tested-healthy rats.Rats were anesthetized with chloral hydrate and placed in the stereotaxic frame(RWD).Two stereotaxic injections of the selective catecholamine neurotoxin 6-OHDA were made unilaterally into the right ascending nigrostriatal dopaminergic pathway using a Hamilton syringe3.The 6-OHDA-induced PD rats divided into five groups:NSCs group,NNSCs group,NNSC+MG group,NNSC+NMG group.Grafted rats were evaluated by rotation test over 30 min after apomorphine administration.4.RT-PCR and Western-blot analysis were used to detect the relative mRNA and protein levels of TH,DAT and Pitx3.5.Immunofluorescence was applied to count the TH+,Iba1_,Nurr1+and DAT+cells in the ventral striatum.Results1.We succeed in constructing the model of Nurrl overexpression in microglia and NSCs.The NSCs were strongly immunopositive for the stem cell marker nestin and the green fluorescent protein;Microglia were strongly immunopositive for the microglia marker CD11b/c and the red fluorescent protein.2.NNSC+NNMG-grafted groups demonstrated a reduction in APO-induced rotation behavior as expressed by the after/before transplant rotational ratio compared to other experimental group.3.Real-time PCR and Western blot showed that compared with other groups,there were much more DAT,TH,neurons in N(NSC+MG)group(P<0.05)in the ventral striatum of PD rats.4.Immunocytobiology showed that compared with other groups,there were much more DAT+,TH+ and Nurrl + neurons in NNSC+NMG groups(P<0.05),and such trend disappeared of Ibal+ cells.Conclusion:These results suggest a potential therapeutic benefit for Nurrl-expressing cells in the treatment of PD.We demonstrate that Microglia and neural stem cell co-transplants with over expression of Nurrl are capable of functionally recovering a PD rat model.