Whole Transcriptomic Study Of Striatum In MPTP-Induced Parkinson's Disease Mice

Objective: The PD-like symptoms of MPTP-induced acute Parkinson's disease(PD)mice will appear in the acute phase and then recover slowly to a certain extent.In addition,PD patients often have retinal involvement,but the molecular mechanism is not clear.In order to study the dynamic changes of transcriptome in striatum of PD mice induced by MPTP were studied by full transcription sequencing,and the differences of transcriptome between striatum and retina were compared.Methords: Thirty 8-week-old mice were randomly divided into three groups: Control group,PD acute model group(D3)and PD model recovery model group(D28).The D3 group and D28 group were modeled with MPTP acute model,and the Control group was injected with saline instead of MPTP.Behavioral tests were carried out by climbing poles and beams.Striatum and retina were taken.High performance liquid chromatography(HPLC)was used to detect dopamine in striatum.Total RNA was extracted from striatum and retina,and rRNA depletion and small RNA library were constructed.Hiseq X-10 platform based on Illumina platform was used to carry out double-ended 150 BP and single-ended 75 BP long reading sequencing,respectively.Long-stranded RNA data were mapped to mouse genome sequence(mm10 version)by RNA STAR software,and differentially expressed genes(DEG)were analyzed by FeatureCount and edgeR.The DEG-enriched gene function and KEGG pathway were analyzed using DAVID and ClueGO plug-ins of CytoScape.lncRNA target genes were predicted by LncTar software,and the interacting lncRNA-mRNA was obtained by combining the expression correlation.SRNAtoolBox was used to analyze differentially expressed microRNAs(DEmiRNA),and TarBase and LncBase databases were used to analyze the binding of DEmiRNA with lncRNA and RNA,and a competitive endogenous RNA(ceRNA)interaction network was constructed.Fluorescence quantitative PCR(qPCR)was used to validate the selected RNA.Results: Behavioral tests confirmed that the PD-like symptoms of the mice first aggravated and then gradually recovered.The content of dopamine in striatum first decreased and then recovered by HPLC.Full transcriptome sequencing showed that the gene expression profiles of D28 mice were similar to those of Control mice,and were significantly different from those of D3 mice.A total of 953 DEGs showed significant changes in relative control in D3 group and tended to recover in D28 group.These protein-coding genes in DEG are mainly concentrated in protein metabolism and neuronal components,as well as ribosome,phosphorylation and Parkinson's disease pathways.Differentially expressed lncRNA(DElncRNA)regulates protein-coding genes,which are mainly concentrated in axon orientation,nervous system development and axon formation,as well as ribosome pathway,focal adhesion,glutamatergic synapse and PD pathway.The expression of DElncRNA such as Neat1 was verified by qPCR.Current studies have shown that Neat1 may play an important role in neurodevelopment and related diseases.We further analyze its ceRNA regulatory network.The results showed that Neat1 may affect histone by regulating mmu-miR-122-5p and collagen by binding to microRNA of mmu-let-7 family.In addition,comparative transcriptome analysis of retina and striatum revealed that both of them expressed PD pathway genes.Of the 129 detected genes,most(117)of them showed relatively similar expression levels.This similar expression pattern may be the potential molecular basis of retinal pathological changes in PD patients.Conclusion: The process of PD-like symptoms appearing and slowly recovering in acute MPTP-induced mice is accompanied by changes in a variety of RNA,especially non-coding RNA,at the transcriptome level.The interaction of ceRNA,especially Neat1,may play a very important role in this process.At the same time,the obvious similarity of PD pathways between striatum and retina may provide new ideas for the research and treatment of PD.