High mobility group box 1 (HMGB1) is implicated in preimplantation embryo development in the mouse.
High mobility group box 1 (HMGB1) regulates multiple cell functions, including transcription, DNA repair, differentiation, and apoptosis. In order to obtain insight into the role of HMGB1 in embryo development, we first evaluated its gene expression levels in mouse preimplantation embryos. Quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) revealed high expression levels in zygotes, and expression steadily increased after zygotic genome activation when normalized to the rabbit Globin mRNA. Indirect immunocytochemistry showed that the HMGB1 protein was also produced in mouse embryos. Injection of a small interfering RNA (siRNA) specific for HMGB1 into zygotes specifically reduced both mRNA expression (P < 0.001) and protein synthesis of HMGB1 in early embryos developed in vitro. Injection of siRNA into the zygote did not affect development to the blastocyst stage, but significantly decreased cell numbers (P < 0.01) in the blastocyst and increased caspase3 (Casp3, P < 0.05) gene expression and apoptosis (P < 0.005). Addition of recombinant HMGB1 (Sigma, H-4652) into the culture medium enhanced the development of zygote stage mouse embryos to blastocysts, in the absence of BSA supplementation. These findings suggest that endogenous and exogenous HMGB1 are implicated in preimplantation embryo development in the mouse.
Cui
Xiang-Shun
XS
National Research Laboratory of Molecular Embryology, Department of Animal Sciences, Chungbuk National University, Seoul, Korea.
Shen
Xing-Hui
XH
Kim
Nam-Hyung
NH
D000818:Animals; D017931:DNA Primers / Q000235:genetics; D004622:Embryo, Mammalian / Q000378:metabolism*; D047108:Embryonic Development / Q000235:genetics*; D018507:Gene Expression Regulation, Developmental / Q000235:genetics*; D024243:HMGB1 Protein / Q000378:metabolism*; D007150:Immunohistochemistry; D020287:In Situ Nick-End Labeling; D016014:Linear Models; D051379:Mice; D020133:Reverse Transcriptase Polymerase Chain Reaction
D017931:DNA Primers; D024243:HMGB1 Protein