| Parkinson’s disease（PD）is a common degenerative disease of the central nervous system in the middle-aged and elderly population,which is mainly manifested by clinical symptoms such as bradykinesia,stiffness,sleep disorders and cognitive impairment.The main pathological features of Parkinson’s disease are the aggregation of Louis bodies and the abnormal number of dopaminergic neurons,which are related to PINK1,the causative gene of Parkinson’s disease.Although numerous studies have revealed that PINK1 plays an important role in the occurrence of Parkinson’s disease,the regulation of PINK1 is unclear.Exploring the regulatory role of PINK1 will provide theoretical guidance for the prevention and treatment of Parkinson’s disease in humans.Drosophila is an important model for the study of neurodegenerative diseases,studies have shown that after the mutation of pink1 homologous gene PINK1 in Drosophila,the life span of Drosophila is significantly shortened,and abnormal apoptosis occurs in dopaminergic neurons in the brain,and due to the dysfunction of mitochondrials,it leads to muscle cell structure degeneration and then causes the crawling ability and flight ability of Drosophila to decline,indicating that the mutation of pink1 in Drosophila can well simulate the symptoms of human Parkinson’s disease.Mbf1 is a class of auxiliary transcription factors that play an important role in the development of the nervous system of Drosophila,and the expression of mbf1 can extend the lifespan of Drosophila to a certain extent,suggesting that Mbf1 may be involved in the regulation of neurodegenerative diseases.Based on the Drosophila model,this study preliminarily explored the expression correlation between Mbf1 and pink1,the regulatory effect of Mbf1 on pink1,and analyzed the functional conservatism of Mbf1 in evolution.The main findings are as follows:（1）Mbf1 regulates the expression of pink1After the mbf1 mutation,the transcription level of homologous genes in multiple PARK signaling pathways,including pink1,in Drosophila showed different degrees of change,and these genes were indirectly or directly affected by Pink1.Overexpression of mbf1 in Drosophila found a significant increase in the transcription level of the pink1 gene.At the same time,the expression levels of mbf1 and pink1 decreased with the age of Drosophila,and the specific spatiotemporal expression of the two was consistent;in in vivo experiments,mbf1 was specifically overexpressed in the brain region of Drosophila,and the expression of pink1 was found to increase significantly.This further illustrates that Mbf1 regulates its expression upstream of pink1.（2）Overexpression of pink1 can rescue the phenotype caused by the mbf1 mutationIn genetic experiments,the mbf1 mutant Drosophila appeared similar to the pink1 mutant,and the number of dopaminergic neurons in the brain of the mbf1 mutant was significantly reduced,the lifespan of the Drosophila was also significantly shortened,and the crawling ability of adults was significantly reduced.However,after overexpression of pink1 in the mbf1 mutation,the phenotype caused by the loss of mbf1 was rescued.This suggests that Mbf1 affects Parkinson’s disease by regulating pink1.（3）Mbf1 is functionally conserved in evolutionUsing the NCBI database to recombine the amino acid sequences of Mbf1 of different species and perform evolutionary analysis,it was found that Mbf1 was highly conserved from yeast to humans.Overexpression of the human Mbf1 homologous protein EDF1（endothelial differentiation related factor 1）in the mbf1 mutant can save the phenotype caused by the reduced crawling ability of the mbf1 mutation,indicating that the two functions are highly conserved.（4）Apt does not participate in the expression regulation of pink1Mbf1 interacts with and regulates pink1 transcription factor as an auxiliary transcription factor is unclear.Apt（Apontic）is a conserved b ZIP transcription factor in the Drosophila nervous system,which is a regulatory element necessary for the development of the Drosophila nervous system,capable of regulating multiple downstream target genes.But whether Apt is involved in the regulation of genes associated with neurodegenerative diseases is unclear.We have shown through Co-IP experiments that Mbf1 and Apt exist in combination.However,knocking down or overexpressing apt in Drosophila’s eye protoplasm,wing primordium,and brain did not have a significant effect on the transcriptional level of pink1.Therefore,it is shown that Apt does not affect the protein expression of pink1.In summary,we found that the highly conserved auxiliary transcription factor Mbf1 affects Parkinson’s disease by regulating the expression of pink1,and we showed that the transcription factor Apt is not involved in regulating the expression of pink1.This study provides a new theoretical basis for the pathogenic mechanism of Parkinson’s disease. |
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