A SENSITIVE REVERSE-TRANSCRIPTASE POLYMERASE CHAIN-REACTION ASSAY FORMEASURING THE EFFECTS OF DEHYDRATION AND GESTATION ON RAT AMOUNTS OF VASOPRESSIN AND OCYTOCIN MESSENGER-RNAS

This study describes a competitive reverse transcriptase polymerase ch ain reaction (RT-PCR) method for assaying the amounts of vasopressin ( AVP) and ocytocin (OT) mRNAs in the rat hypothalamus and uterus. Despi te the low concentrations of these mRNAs, the RT-PCR method readily me asured both AVP and OT mRNAs in the same sample. A common internal sta ndard for both reactions was designed to quantify the reaction. Both A VP and OT mRNAs were readily quantified in a 75 ng sample of total RNA from the hypothalamus. Water deprivation stimulated AVP mRNA producti on 3-fold and OT mRNA production 1.7-fold in the hypothalamus. Gestati on only influenced the amount of OT mRNA in the hypothalamus (3-fold i ncrease) and uterus (38-fold increase). The amount of AVP mRNA in the hypothalamus remained unchanged and no AVP mRNA was detected in the ut eri of either non-pregnant or pregnant rats. This competitive RT-PCR i s a powerful tool that provides rapid and precise assays, of AVP and O T mRNAs. (C) 1997 Elsevier Science Ireland Ltd.