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Journal of Assisted Reproduction and Genetics

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01-06-2015 | Embryo Biology

Dysfunction in gap junction intercellular communication induces aberrant behavior of the inner cell mass and frequent collapses of expanded blastocysts in mouse embryos

Authors: Kazue Togashi, Jin Kumagai, Emiko Sato, Hiromitsu Shirasawa, Yuki Shimoda, Kenichi Makino, Wataru Sato, Yukiyo Kumazawa, Yasufumi Omori, Yukihiro Terada

Published in: Journal of Assisted Reproduction and Genetics | Issue 6/2015

Abstract

Purpose

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).

Methods

ICR mouse embryos were exposed to two types of GJis in CZB medium: oleamide (0 to 50 μM) 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.

Results

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.

Conclusions

GJIC plays an important role in blastocyst formation, collapses of expanded blastocysts, and the ICM construction in mouse embryos.

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Title: Dysfunction in gap junction intercellular communication induces aberrant behavior of the inner cell mass and frequent collapses of expanded blastocysts in mouse embryos
Authors: Kazue Togashi
Jin Kumagai
Emiko Sato
Hiromitsu Shirasawa
Yuki Shimoda
Kenichi Makino
Wataru Sato
Yukiyo Kumazawa
Yasufumi Omori
Yukihiro Terada
Publication date: 01-06-2015
Publisher: Springer US
Published in: Journal of Assisted Reproduction and Genetics / Issue 6/2015
Print ISSN: 1058-0468
Electronic ISSN: 1573-7330
DOI: https://doi.org/10.1007/s10815-015-0479-1

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Dysfunction in gap junction intercellular communication induces aberrant behavior of the inner cell mass and frequent collapses of expanded blastocysts in mouse embryos

Abstract

Purpose

Methods

Results

Conclusions

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusions

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