PMID- 33196062
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20201118
VI  - 3
IP  - 2
DP  - 2020
TI  - Comparison of rat fetal sex determination using placental gDNA and mRNA via 
      qRT-PCR.
LID - 107 [pii]
AB  - A growing need exists to consider fetal sex as a biological variable and 
      accurately assess sex-specific effects. Among the multiple methods used to 
      determine fetal sex, quantitative real-time polymerase chain reaction (qRT-PCR) 
      of Sry (sex-determining region Y) with genomic DNA (gDNA) is commonly used in 
      addition to use of methodologies such as transcriptomics and detection of Barr 
      body. However, Sry messenger RNA (mRNA), a product of Sry gDNA, has not been 
      previously assessed for sex determination. Using placental samples from 
      timed-pregnant Wistar rats at gestational day (GD) 16, this study assessed the 
      compatibility of Sry detection using gDNA versus mRNA to determine fetal sex. 
      Samples used in this current study come from a larger study that investigated 
      trichloroethylene (TCE) reproductive toxicity and potential modulation by 
      N-acetyl-L-cysteine (NAC) and aminooxyacetic acid (AOAA). In 90 out of 91 
      samples, the sex classification determined by gDNA matched the sex classification 
      determined by mRNA analyzing Sry (Sry/B2m) values. For both gDNA and mRNA, 
      statistically significant differences in Sry/B2m values between males and females 
      were observed with samples considered in totality and when samples were separated 
      by treatment groups (all comparisons were p<0.01 or below, and all but two 
      comparisons were p<0.001 or below). Finally, the validity of using Sry Cq values 
      to determine fetal sex and the B2m reference gene were also discussed. Together, 
      this study suggests that determination of fetal sex in Wistar rats can be 
      accomplished using Sry measurements in gDNA or mRNA with highly compatible 
      results.
FAU - Su, Anthony L
AU  - Su AL
AUID- ORCID: 0000-0002-5280-6423
AD  - Department of Environmental Health Sciences, School of Public Health, University 
      of Michigan, Ann Arbor, MI 48109-2029 USA.
AD  - Current address: Perelman School of Medicine, University of Pennsylvania, 
      Philadelphia, PA 19104 USA.
FAU - Loch-Caruso, Rita
AU  - Loch-Caruso R
AUID- ORCID: 0000-0002-5993-2799
AD  - Department of Environmental Health Sciences, School of Public Health, University 
      of Michigan, Ann Arbor, MI 48109-2029 USA.
LA  - eng
GR  - P30 ES017885/ES/NIEHS NIH HHS/United States
GR  - P42 ES017198/ES/NIEHS NIH HHS/United States
GR  - T32 ES007062/ES/NIEHS NIH HHS/United States
GR  - T32 HD079342/HD/NICHD NIH HHS/United States
PT  - Journal Article
DEP - 20200705
PL  - England
TA  - J Mol Biol Methods
JT  - Journal of molecular biology and methods
JID - 101772754
PMC - PMC7665159
MID - NIHMS1640755
COIS- Competing Interests Statement The authors have no competing interests to declare.
EDAT- 2020/11/17 06:00
MHDA- 2020/11/17 06:01
PMCR- 2020/11/13
CRDT- 2020/11/16 08:57
PHST- 2020/11/16 08:57 [entrez]
PHST- 2020/11/17 06:00 [pubmed]
PHST- 2020/11/17 06:01 [medline]
PHST- 2020/11/13 00:00 [pmc-release]
AID - 107 [pii]
PST - ppublish
SO  - J Mol Biol Methods. 2020;3(2):107. Epub 2020 Jul 5.