Open AccessA DNA Methyltransferase Inhibitor, 5‐Aza‐2′‐Deoxycytidine, Exacerbates Neurotoxicity and Upregulates P arkinson's Disease‐Related Genes in Dopaminergic Neurons
Author(s) -
Wang Yong,
Wang Xuan,
Li Ran,
Yang ZhaoFei,
Wang YiZheng,
Gong XiaoLi,
Wang XiaoMin
Publication year - 2013
Publication title -
cns neuroscience and therapeutics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.403
H-Index - 69
eISSN - 1755-5949
pISSN - 1755-5930
DOI - 10.1111/cns.12059
Subject(s) - dopaminergic , neurotoxicity , dna demethylation , dna methyltransferase , tyrosine hydroxylase , chemistry , rotenone , viability assay , microbiology and biotechnology , methyltransferase , biology , pharmacology , dna methylation , apoptosis , dopamine , biochemistry , gene expression , methylation , gene , mitochondrion , endocrinology , enzyme , toxicity , organic chemistry
Summary Aims To investigate effects of DNA methyltransferase ( DNMT ) inhibitors on dopaminergic neurons and its underlied mechanism. Methods The DNMT inhibitor 5‐aza‐2′‐deoxycytidine (5‐aza‐ dC ) was tested in cultured dopaminergic cells. Cell viability and apoptosis were assayed with 5‐aza‐ dC alone. Neurotoxicity of 1‐methyl‐4‐phenylpyridinium ( MPP + ), 6‐hydroxydopamine or rotenone was also assayed with 5‐aza‐ dC pretreatment. And mRNA levels of several key PD ‐related genes were examined by semiquantitative RT ‐ PCR . Furthermore, C p G methylation of α‐synuclein promoter was examined by bisulfite sequencing. Results 5‐aza‐ dC resulted in decreased cell viability and increased apoptosis in dopaminergic neuronal cells. Pretreatment with 5‐aza‐ dC exacerbated neurotoxic damage to dopaminergic neurons induced by MPP + , 6‐hydroxydopamine or rotenone. 5‐aza‐ dC also induced transcriptional upregulation of the key PD ‐related genes tyrosine hydroxylase and α‐synuclein . And demethylation of C p G in α‐synuclein promoter was also induced by 5‐aza‐ dC and MPP + . Conclusions This DNMT inhibitor might influence pathogenesis of PD . And demethylation induced by DNMT inhibitor might contribute to dopaminergic neuron death, by increasing vulnerability of dopaminergic neurons to neurotoxins and by misregulating transcription of key PD ‐related genes. Our data also suggested DNMT inhibitors may cause multiple effects on dopaminergic neurons.