期刊
JOURNAL OF ASSISTED REPRODUCTION AND GENETICS
卷 32, 期 6, 页码 969-976出版社
SPRINGER/PLENUM PUBLISHERS
DOI: 10.1007/s10815-015-0479-1
关键词
Gap junction inhibitor; Time-lapse movie observation; Delayed blastocyst formation; Blastocoel collapse; ICM division
资金
- JSPS KAKENHI [25462549]
- National Centre for Child Health and Development [24-6]
- Grants-in-Aid for Scientific Research [25462549, 24590472, 15K15593] Funding Source: KAKEN
We investigated the role of gap junctions (GJs) in embryological differentiation, and observed the morphological behavior of the inner cell mass (ICM) by time-lapse movie observation (TLM) with gap junction inhibitors (GJis). ICR mouse embryos were exposed to two types of GJis in CZB medium: oleamide (0 to 50 mu M) and 1-heptanol (0 to 10 mM). We compared the rate of blastocyst formation at embryonic day 4.5 (E4.5) with E5.5. We also observed and evaluated the times from the second cleavage to each embryonic developing stage by TLM. We investigated embryonic distribution of DNA, Nanog protein, and Connexin 43 protein with immunofluorescent staining. In the comparison of E4.5 with E5.5, inhibition of gap junction intercellular communication (GJIC) delayed embryonic blastocyst formation. The times from the second cleavage to blastocyst formation were significantly extended in the GJi-treated embryos (control vs with oleamide, 2224 +/- 179 min vs 2354 +/- 278 min, p = 0.013). Morphological differences were traced in control versus GJi-treated embryos until the hatching stage. Oleamide induced frequent severe collapses of expanded blastocysts (77.4 % versus 26.3 %, p = 0.0001) and aberrant ICM divisions connected to sticky strands (74.3 % versus 5.3 %, p = 0.0001). Immunofluorescent staining indicated Nanog-positive cells were distributed in each divided ICM. GJIC plays an important role in blastocyst formation, collapses of expanded blastocysts, and the ICM construction in mouse embryos.
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