{"title":"Comparison of rat fetal sex determination using placental gDNA and mRNA via qRT-PCR.","authors":"Anthony L Su, Rita Loch-Caruso","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>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 <i>Sry</i> (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, <i>Sry</i> messenger RNA (mRNA), a product of <i>Sry</i> 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 <i>Sry</i> 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 <i>Sry</i> (<i>Sry</i>/<i>B2m</i>) values. For both gDNA and mRNA, statistically significant differences in <i>Sry</i>/<i>B2m</i> values between males and females were observed with samples considered in totality and when samples were separated by treatment groups (all comparisons were <i>p</i><0.01 or below, and all but two comparisons were <i>p</i><0.001 or below). Finally, the validity of using <i>Sry</i> Cq values to determine fetal sex and the <i>B2m</i> reference gene were also discussed. Together, this study suggests that determination of fetal sex in Wistar rats can be accomplished using <i>Sry</i> measurements in gDNA or mRNA with highly compatible results.</p>","PeriodicalId":93119,"journal":{"name":"Journal of molecular biology and methods","volume":"3 2","pages":""},"PeriodicalIF":0.0,"publicationDate":"2020-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7665159/pdf/nihms-1640755.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38607731","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}