Sexing of postimplantation rat embryos in stored two-dimensional electrophoresis (2-DE) samples by polymerase chain reaction (PCR) of an Sry sequence.
Proteomic analysis of developmental toxicity by two-dimensional electrophoresis (2-DE) may detect gender-related toxic effects in embryos without visible gender characteristics. In the present study, we explored sexing of rat embryo stored in frozen 2-DE samples by polymerase chain reaction (PCR) of a male-specific gene sequence, sex determining region Y (Sry). The embryo proper and yolk sac membrane at gestation day 11 from Wistar rats were used for stored embryonic 2-DE samples. The embryonic 2-DE samples were desalted and their total DNA was extracted. The Sry sequence in the extracted DNA was amplified by PCR and the product was analyzed by agarose gel electrophoresis. The embryos with the PCR product of Sry were determined as male, and those without the product were determined as female. It was concluded that stored embryonic 2-DE samples could be used for retrospective examination of gender-related effects in proteomic analysis of developmental toxicity.
Miyajima
Atsuko
A
Sunouchi
Momoko
M
Mitsunaga
Katsuyoshi
K
Yamakoshi
Yoko
Y
Nakazawa
Ken
K
Usami
Makoto
M
D000818:Animals; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D015180:Electrophoresis, Gel, Two-Dimensional; D046149:Embryo Culture Techniques; D010064:Embryo Implantation; D004622:Embryo, Mammalian / Q000196:embryology*; D047108:Embryonic Development; D005260:Female; D024282:Genes, sry / Q000235:genetics*; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results; D017422:Sequence Analysis, DNA; D019849:Sex Determination Processes
D004247:DNA