Quantification of thioredoxin mRNA expression in the rat hippocampus by real-time PCR following oxidative stress.
Thioredoxin (Trx) is a multifunctional protein with a redox-active disulfide/dithiol in the active site. Thioredoxin, with its redox-regulating and reactive oxygen species (ROS) scavenging activities, plays several important biologic roles both in intracellular and extracellular compartments. The purpose of this report was to quantify the relative expression of Trx in rat hippocampus following an oxidative stress-involving treatment such as kainic acid (KA) using real-time PCR and the 2(-DeltaDeltaC(T)) method. The relative changes in expression of Trx mRNA in KA-treated and control animals were significantly different as 2.02 +/- 0.77 and 1.0 +/- 0.26, respectively (P<0.05). Minimum and maximum n-fold changes in Trx expression in KA-treated and control animals were determined as (1.4-5.2) and (0.8-1.3), respectively. Thus, real-time PCR and the 2(-DeltaDeltaC(T)) method for data analysis from real-time PCR were found to be an accurate and sensitive method for quantifying Trx mRNA levels.
Yalcin
Ayfer
A
Department of Biochemistry, Faculty of Pharmacy, Ege University, Izmir, Turkey. yalcina@pharm.ege.edu.tr
D000818:Animals; D001483:Base Sequence; D015870:Gene Expression; D006624:Hippocampus / Q000378:metabolism*; D007608:Kainic Acid / Q000494:pharmacology; D008297:Male; D008969:Molecular Sequence Data; D018384:Oxidative Stress; D016133:Polymerase Chain Reaction / Q000379:methods*; D012333:RNA, Messenger / Q000032:analysis* / Q000378:metabolism; D051381:Rats; D013879:Thioredoxins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*
D012333:RNA, Messenger; D013879:Thioredoxins; D007608:Kainic Acid