Quantitative RT-PCR assay of 5-HT1A and 5-HT2A serotonin receptor mRNAs using genomic DNA as an external standard.
Brain serotonin 5-HT(1A) and 5-HT(2A) receptors have been implicated in both normal and pathological behavior, and in the action of anxiolytic and antidepressant drugs. In this study, detailed description and verification of a new RT-PCR technique to quantify the number of copies of 5-HT(1A) and 5-HT(2A) receptor mRNAs in the brain is presented. The number of copies of beta-actin and 5-HT(1A) or 5-HT(2A) receptor mRNAs in rat brain samples was evaluated with respect to the genomic DNA solution as the external exogenous standard. The expression of 5-HT receptors was calculated as the number of receptor mRNA copies per 100 copies of corresponding beta-actin mRNA. This presented technique is reliable, simple and can be easily set up in any neurobiological laboratory.
Kulikov
Alexander V
AV
Institute of Cytology and Genetics, Russian Academy of Sciences, Avenue Lavrentyev, 10 630090 Novosibirsk, Russia. akulikov@ngs.ru
Naumenko
Vladimir S
VS
Voronova
Irina P
IP
Tikhonova
Maria A
MA
Popova
Nina K
NK
D000199:Actins / Q000032:analysis; D000818:Animals; D001923:Brain Chemistry / Q000502:physiology; D004247:DNA / Q000032:analysis*; D016678:Genome; D008297:Male; D009448:Neurochemistry / Q000379:methods*; D012333:RNA, Messenger / Q000032:analysis*; D051381:Rats; D017208:Rats, Wistar; D044282:Receptor, Serotonin, 5-HT1A / Q000235:genetics*; D044402:Receptor, Serotonin, 5-HT2A / Q000235:genetics*; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012701:Serotonin / Q000378:metabolism
D000199:Actins; D012333:RNA, Messenger; D044402:Receptor, Serotonin, 5-HT2A; D044282:Receptor, Serotonin, 5-HT1A; D012701:Serotonin; D004247:DNA