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. 2013;59(2):151-8.
doi: 10.1262/jrd.2012-122. Epub 2012 Dec 20.

Changes in the expression patterns of the genes involved in the segregation and function of inner cell mass and trophectoderm lineages during porcine preimplantation development

Takashi Fujii  1 , Nobuyuki Sakurai, Tsubasa Osaki, Gentaro Iwagami, Hiroki Hirayama, Akira Minamihashi, Tsutomu Hashizume, Ken Sawai
Affiliations

Changes in the expression patterns of the genes involved in the segregation and function of inner cell mass and trophectoderm lineages during porcine preimplantation development

Takashi Fujii et al. J Reprod Dev. 2013.

Abstract

In mouse embryos, segregation of the inner cell mass (ICM) and trophectoderm (TE) lineages is regulated by genes, such as OCT-4, CDX2 and TEAD4. However, the molecular mechanisms that regulate the segregation of the ICM and TE lineages in porcine embryos remain unknown. To obtain insights regarding the segregation of the ICM and TE lineages in porcine embryos, we examined the mRNA expression patterns of candidate genes, OCT-4, CDX2, TEAD4, GATA3, NANOG, FGF4, FGFR1-IIIc and FGFR2-IIIc, in blastocyst and elongated stage embryos. In blastocyst embryos, the expression levels of OCT-4, FGF4 and FGFR1-IIIc were significantly higher in the ICM than in the TE, while the CDX2, TEAD4 and GATA3 levels did not differ between the ICM and TE. The expression ratio of CDX2 to OCT-4 (CDX2/OCT-4) also did not differ between the ICM and TE at the blastocyst stage. In elongated embryos, OCT-4, NANOG, FGF4 and FGFR1-IIIc were abundantly expressed in the embryo disc (ED; ICM lineage), but their expression levels were very low in the TE. In contrast, the CDX2, TEAD4 and GATA3 levels were significantly higher in the TE than in the ED. In addition, the CDX2/OCT-4 ratio was markedly higher in the TE than in the ED. We demonstrated that differences in the expression levels of OCT-4, CDX2, TEAD4, GATA3, NANOG, FGF4, FGFR1-IIIc and FGFR2-IIIc genes between ICM and TE lineages cells become more clear during development from porcine blastocyst to elongated embryos, which indicates the possibility that in porcine embryos, functions of ICM and TE lineage cells depend on these gene expressions proceed as transition from blastocyst to elongated stage.

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Figures

Fig. 1.
Fig. 1.
Representative photographs of porcine blastocyst (A, B) and elongated stage embryos (C, ovoid; D, filamentous). Scale bars represent 2 mm (C) and 2.5 mm (D). Arrows indicate the ICM (A, B) or ED (C, D) in embryos. Arrowheads indicate the TE in embryos (A, B, C, D).
Fig. 2.
Fig. 2.
Relative abundance (mean ± SE) of (A) OCT-4, (B) CDX2, (D) TEAD4, (E) GATA3, (F) FGF4, and (G) FGFR1-IIIc transcripts in porcine blastocyst embryos (n = 5). (C) The expression ratio of CDX2 to OCT-4 mRNAs (CDX2/OCT-4) in porcine blastocyst embryos (n = 5). Expression levels of OCT-4, CDX2, TEAD4, GATA3, FGF4 and FGFR1-IIIc mRNAs were normalized to GAPDH expression. a,b Different superscripts indicate a significant difference (P<0.01–0.05). NANOG and FGFR2-IIIc mRNA were not detected in porcine blastocyst embryos.
Fig. 3.
Fig. 3.
Relative abundance (mean ± SE) of (A) OCT-4, (B) CDX2, (D) TEAD4, (E) GATA3, (F) NANOG, (G) FGF4, (H) FGFR1-IIIc and (I) FGFR2-IIIc transcripts in porcine elongated embryos (ovoid [n = 4] and filamentous [n = 5]). (C) The expression ratio of CDX2 to OCT-4 mRNAs (CDX2/OCT-4) in porcine elongated embryos (ovoid [n = 4] and filamentous [n = 5]). The bars for the CDX2/OCT-4 ratio represent a log scale. Expression levels of OCT-4, CDX2, TEAD4, GATA3, NANOG, FGF4, FGFR1-IIIc and FGFR2-IIIc mRNAs were normalized to GAPDH expression. a,b,c Different superscripts indicate a significant difference (P<0.01–0.05).
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