title: Methamphetamine alters reference gene expression in nigra and striatum of adult rat brain.
abstract: The nigrostriatal dopaminergic system is a major lesion target for methamphetamine (MA), one of the most addictive and neurotoxic drugs of abuse. High doses of MA alter the expression of a large number of genes. Reference genes (RGs) are considered relatively stable and are often used as standards for quantitative real-time PCR (qRT-PCR) reactions. The purpose of this study was to determine whether MA altered the expression of RGs and to identify the appropriate RGs for gene expression studies in animals receiving MA. Adult male Sprague-Dawley rats were treated with high doses of MA or saline. Striatum and substantia nigra were harvested at 2h or 24h after MA administration. The expression and stability of 10 commonly used RGs were examined using qRT-PCR and then evaluated by geNorm and Normfinder. We found that MA altered the expression of selected RGs. These candidate RGs presented differential stability in the striatum and in substantia nigra at both 2h and 24h after MA injection. Selection of an unstable RG as a standard altered the significance of tyrosine hydroxylase (TH) mRNA expression after MA administration. In conclusion, our data show that MA site- and time-dependently altered the expression of RGs in nigrostriatal dopaminergic system. These temporal and spatial factors should be considered when selecting appropriate RGs for interpreting the expression of target genes in animals receiving MA.
authors.affiliation: Neural Protection and Regeneration Section, National Institute on Drug Abuse, Intramural Research Program, NIH, Baltimore, MD 21224, USA. Electronic address: yi.he2@nih.gov.
authors.forename: Yi
authors.initials: Y
authors.lastname: He
authors.forename: Seongjin
authors.initials: S
authors.lastname: Yu
authors.forename: Eunkyung
authors.initials: E
authors.lastname: Bae
authors.forename: Hui
authors.initials: H
authors.lastname: Shen
authors.forename: Yun
authors.initials: Y
authors.lastname: Wang
mesh_terms: D000818:Animals; D000697:Central Nervous System Stimulants / Q000494:pharmacology*; D003342:Corpus Striatum / Q000187:drug effects*; D005786:Gene Expression Regulation / Q000187:drug effects*; D008297:Male; D008694:Methamphetamine / Q000494:pharmacology*; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013378:Substantia Nigra / Q000187:drug effects*; D014446:Tyrosine 3-Monooxygenase / Q000235:genetics / Q000378:metabolism*
chemical_list: D000697:Central Nervous System Stimulants; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D008694:Methamphetamine; D014446:Tyrosine 3-Monooxygenase
keywords: Housekeeping genes; Methamphetamine; Nigra; Real-time PCR; Reference genes; Striatum
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