A sensitive reverse transcriptase polymerase chain reaction assay for measuring the effects of dehydration and gestation on rat amounts of vasopressin and ocytocin mRNAs.
This study describes a competitive reverse transcriptase polymerase chain reaction (RT-PCR) method for assaying the amounts of vasopressin (AVP) and ocytocin (OT) mRNAs in the rat hypothalamus and uterus. Despite the low concentrations of these mRNAs, the RT-PCR method readily measured both AVP and OT mRNAs in the same sample. A common internal standard for both reactions was designed to quantify the reaction. Both AVP and OT mRNAs were readily quantified in a 75 ng sample of total RNA from the hypothalamus. Water deprivation stimulated AVP mRNA production 3-fold and OT mRNA production 1.7-fold in the hypothalamus. Gestation only influenced the amount of OT mRNA in the hypothalamus (3-fold increase) and uterus (38-fold increase). The amount of AVP mRNA in the hypothalamus remained unchanged and no AVP mRNA was detected in the uteri of either non-pregnant or pregnant rats. This competitive RT-PCR is a powerful tool that provides rapid and precise assays of AVP and OT mRNAs.
LeMoullec
J M
JM
Roussel Uclaf, Romainville, France.
Jouquey
S
S
Corvol
P
P
Pinet
F
F
D000818:Animals; D001127:Arginine Vasopressin / Q000096:biosynthesis* / Q000235:genetics; D001483:Base Sequence; D004247:DNA / Q000737:chemistry; D017931:DNA Primers; D003681:Dehydration / Q000378:metabolism*; D005260:Female; D007031:Hypothalamus / Q000378:metabolism*; D008297:Male; D008969:Molecular Sequence Data; D010121:Oxytocin / Q000096:biosynthesis* / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods; D011247:Pregnancy; D011270:Pregnancy, Animal / Q000378:metabolism*; D018075:RNA, Complementary; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D014158:Transcription, Genetic; D014599:Uterus / Q000378:metabolism*
D017931:DNA Primers; D018075:RNA, Complementary; D012333:RNA, Messenger; D001127:Arginine Vasopressin; D010121:Oxytocin; D004247:DNA