CYP2C76 deficiency is embryonic lethal in cynomolgus macaques: The potential role of CYP2C76 in early embryogenesis.
Cynomolgus macaques are an important primate species for drug metabolism studies; however cynomolgus CYP2C76, an important drug-metabolizing enzyme, accounts for drug metabolism differences to humans, so that CYP2C76-null animals might show drug-metabolizing properties more similar to humans. In this study, attempts were made to produce CYP2C76-null animals by assisted reproduction technology. Oocytes and sperm collected from the heterozygotes for the null allele (c.449TG > A) were subjected to intracytoplasmic sperm injection, and the embryos produced were cultured in vitro through the blastocyst stage. Preimplantation genetic diagnosis using a biopsied portion of the blastocyst revealed that none of the 32 blastocysts analyzed were homozygotes. In contrast, 2 of the 20 embryos analyzed were homozygotes at the 8-cell stage, indicating that CYP2C76-null embryos most likely stop developing between the 8-cell and blastocyst stage. By polymerase chain reaction, expression of CYP2C76 mRNA was detected in oocytes and blastocysts, but not in 2-, 4-, 8-, or 16/32-cell stage embryos. Metabolic assays showed that CYP2C76 metabolized progesterone. These results indicated that CYP2C76 null was likely embryonic lethal, suggesting its potential role during early embryogenesis in cynomolgus macaques.
Drug Safety Research Center (DSR), Shin Nippon Biomedical Laboratories (SNBL), Ltd., Kagoshima, Japan; SNBL USA, Ltd., Everett, WA, USA.
Shuzo
S
Koyama
Drug Safety Research Center (DSR), Shin Nippon Biomedical Laboratories (SNBL), Ltd., Kagoshima, Japan; SNBL USA, Ltd., Everett, WA, USA.
Koji
K
Fukuda
Drug Safety Research Center (DSR), Shin Nippon Biomedical Laboratories (SNBL), Ltd., Kagoshima, Japan; SNBL USA, Ltd., Everett, WA, USA.
Sho
S
Watanabe
Drug Safety Research Center (DSR), Shin Nippon Biomedical Laboratories (SNBL), Ltd., Kagoshima, Japan; Pharmacokinetics and Bioanalysis Center (PBC), Shin Nippon Biomedical Laboratories (SNBL), Ltd., Kainan, Japan.
Akinori
A
Matsushita
Research Center for Animal Life Science, Shiga University of Medical Science, Otsu, Japan.
Hideaki
H
Tsuchiya
Graduate School of Biology-Oriented Science and Technology, Kinki University, Kinokawa, Japan.
Nahoko
N
Fujinami
Pharmacokinetics and Bioanalysis Center (PBC), Shin Nippon Biomedical Laboratories (SNBL), Ltd., Kainan, Japan.
Sakae
S
Kohara
Laboratory of Drug Metabolism and Pharmacokinetics, Showa Pharmaceutical University, Machida, Japan.
Norie
N
Murayama
Laboratory of Theriogenology, Department of Veterinary Clinical Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Japan.
Masashi
M
Nagano
Laboratory of Drug Metabolism and Pharmacokinetics, Showa Pharmaceutical University, Machida, Japan.
Hiroshi
H
Yamazaki
Drug Safety Research Center (DSR), Shin Nippon Biomedical Laboratories (SNBL), Ltd., Kagoshima, Japan; SNBL USA, Ltd., Everett, WA, USA.
Koichiro
K
Fukuzaki
Pharmacokinetics and Bioanalysis Center (PBC), Shin Nippon Biomedical Laboratories (SNBL), Ltd., Kainan, Japan. Electronic address: uno-yasuhiro@snbl.co.jp.
Yasuhiro
Y
Uno
Graduate School of Biology-Oriented Science and Technology, Kinki University, Kinokawa, Japan. Electronic address: Hosoi@waka.kindai.ac.jp.
Yoshihiko
Y
Hosoi
D000818:Animals; D003577:Cytochrome P-450 Enzyme System / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism*; D020964:Embryo Loss / Q000235:genetics*; D047108:Embryonic Development; D005260:Female; D008252:Macaca fascicularis / Q000196:embryology* / Q000235:genetics*; D008297:Male; D009865:Oocytes / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013094:Spermatozoa / Q000378:metabolism
D012333:RNA, Messenger; C400322:cytochrome P-450 CYP2C subfamily; D003577:Cytochrome P-450 Enzyme System
Cynomolgus macaque; Cytochrome P450; Embryogenesis; Null allele; Progesterone