Quantitative RT-PCR assay of 5-HT1A and 5-HT2A serotonin receptor mRNAs using genomic DNA as an external standard

J Neurosci Methods. 2005 Jan 30;141(1):97-101. doi: 10.1016/j.jneumeth.2004.06.005.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / analysis
  • Animals
  • Brain Chemistry / physiology
  • DNA / analysis*
  • Genome
  • Male
  • Neurochemistry / methods*
  • RNA, Messenger / analysis*
  • Rats
  • Rats, Wistar
  • Receptor, Serotonin, 5-HT1A / genetics*
  • Receptor, Serotonin, 5-HT2A / genetics*
  • Reference Standards
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Serotonin / metabolism

Substances

  • Actins
  • RNA, Messenger
  • Receptor, Serotonin, 5-HT2A
  • Receptor, Serotonin, 5-HT1A
  • Serotonin
  • DNA