Comparison of the time courses of selective gene expression and dopaminergic depletion induced by MPP+ in MN9D cells.
Parkinson's disease (PD) is a common neurodegenerative disease characterized by progressive loss of midbrain dopaminergic neurons with unknown etiology. MPP+ (1-methyl-4-phenylpyridinium ion) is the active metabolite of the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), which induces Parkinson's-like symptoms in humans and animals. MPTP/MPP+ produces selective dopaminergic neuronal degeneration, therefore, these agents are commonly used to study the pathogenesis of PD. However, the mechanisms of their toxicity have not been fully elucidated. Recently, we reported in a microarray study using a midbrain-derived dopaminergic neuronal cell line, MN9D, that MPP+ induced significant changes in a number of genes known to be associated with the dopaminergic system. In this study, we investigated the expression time courses of six genes using real-time RT-PCR, and compared them with the progressive dopaminergic depletion caused by MPP+. Our data showed that dopamine content was significantly decreased after 0.5h of MPP+ (200 microM) exposure and was completely depleted after 40 h. The expression of Gpr37, which is closely related to the pathogenesis of autosomal recessive juvenile Parkinsonism, was up-regulated after 0.5h, and stayed up-regulated up to 48 h. Txnip, which is critical to the adjustment of cellular redox status, was down-regulated after 1h and stayed down-regulated up to 48 h. Ldh1 and Cdo1, which are also involved in oxidative stress, were down-regulated after 16 h and stayed down-regulated up to 48 h. Two pro-apoptotic genes, Egln3 and Bnip3, were down-regulated after 2 and 4h, and stayed down-regulated up to 48 h. These findings suggested that the time course of expression for multiple genes correlated with the dopaminergic depletion; and MPP+-induced neurotoxicity in MN9D cells could be used as a model to further explore the roles of these and other genes in the pathogenesis and possible treatment of PD.
Wang
Jianyong
J
Neurochemistry Laboratory, Division of Neurotoxicology, National Center for Toxicological Research, Jefferson, AR 72079, USA. jianyong.wang@fda.hhs.gov <jianyong.wang@fda.hhs.gov>
Duhart
Helen M
HM
Xu
Zengjun
Z
Patterson
Tucker A
TA
Newport
Glenn D
GD
Ali
Syed F
SF
D015655:1-Methyl-4-phenylpyridinium / Q000633:toxicity*; D000818:Animals; D002352:Carrier Proteins / Q000235:genetics; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000235:genetics; D018450:Disease Progression; D004298:Dopamine / Q000172:deficiency*; D015536:Down-Regulation / Q000187:drug effects / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects* / Q000235:genetics; D006540:Herbicides / Q000633:toxicity; D064799:Hypoxia-Inducible Factor-Proline Dioxygenases; D017874:Immediate-Early Proteins / Q000235:genetics; D007527:Isoenzymes / Q000235:genetics; D007770:L-Lactate Dehydrogenase / Q000235:genetics; D008565:Membrane Proteins / Q000235:genetics; D051379:Mice; D024101:Mitochondrial Proteins / Q000235:genetics; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism; D020734:Parkinsonian Disorders / Q000235:genetics* / Q000378:metabolism / Q000503:physiopathology; D012333:RNA, Messenger / Q000187:drug effects / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled / Q000235:genetics; D013378:Substantia Nigra / Q000187:drug effects / Q000378:metabolism / Q000503:physiopathology; D013879:Thioredoxins / Q000235:genetics; D013997:Time Factors
C508686:BNip3 protein, mouse; D002352:Carrier Proteins; D004268:DNA-Binding Proteins; C509804:Gpr37 protein, mouse; D006540:Herbicides; D017874:Immediate-Early Proteins; D007527:Isoenzymes; D008565:Membrane Proteins; D024101:Mitochondrial Proteins; D012333:RNA, Messenger; D043562:Receptors, G-Protein-Coupled; C445672:Txnip protein, mouse; D013879:Thioredoxins; D007770:L-Lactate Dehydrogenase; C409667:lactate dehydrogenase 1; C087100:Egln3 protein, rat; D064799:Hypoxia-Inducible Factor-Proline Dioxygenases; D015655:1-Methyl-4-phenylpyridinium; D004298:Dopamine