An RNA spiking method demonstrates that 18S rRNA is regulated by progesterone in the mouse uterus.
Identifying suitable housekeeping genes for quantitative RT-PCR in the uterus is problematic, as this tissue undergoes significant structural and functional alterations during the oestrous cycle and pregnancy in response to circulating hormones. The suitability of 18S rRNA as a housekeeping gene in mouse uterus was investigated by introducing an 'RNA spike' standard into the reverse transcription reaction. 18S rRNA levels increased by Day 4 of pregnancy and after progesterone administration in ovariectomized mice. We conclude that 18S rRNA is not a suitable housekeeping gene for quantitative RT-PCR analysis in progesterone-responsive tissues, and the RNA spiking method provides a suitable alternative.
Craythorn
R G
RG
Centre for Reproduction and Development, Monash Institute of Medical Research, Monash University, 27-31 Wright St, Clayton, VIC 3168, Australia.
Girling
J E
JE
Hedger
M P
MP
Rogers
P A W
PA
Winnall
W R
WR
D000818:Animals; D004958:Estradiol / Q000494:pharmacology; D005260:Female; D051379:Mice; D010052:Ovariectomy; D011247:Pregnancy; D011374:Progesterone / Q000378:metabolism* / Q000494:pharmacology; D012337:RNA, Ribosomal, 18S / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D014599:Uterus / Q000187:drug effects / Q000378:metabolism*
D012337:RNA, Ribosomal, 18S; D011374:Progesterone; D004958:Estradiol