Ultrastructural demonstration of Cx43 gap junctions in induced pluripotent stem cells from human cord blood

Beckmann, Anja; Schubert, Madline; Hainz, Nadine; Haase, Alexandra; Martin, Ulrich; Tschernig, Thomas; Meier, Carola

doi:10.1007/s00418-016-1469-9

Original Paper
Published: 25 July 2016

Ultrastructural demonstration of Cx43 gap junctions in induced pluripotent stem cells from human cord blood

Anja Beckmann¹,
Madline Schubert^2,3,
Nadine Hainz¹,
Alexandra Haase^2,3,
Ulrich Martin^2,3,
Thomas Tschernig¹ &
Carola Meier¹

Histochemistry and Cell Biology volume 146, pages 529–537 (2016)Cite this article

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Abstract

Gap junction proteins are essential for direct intercellular communication but also influence cellular differentiation and migration. The expression of various connexin gap junction proteins has been demonstrated in embryonic stem cells, with Cx43 being the most intensely studied. As Cx43 is the most prominent gap junction protein in the heart, cardiomyocyte-differentiated stem cells have been studied intensely. To date, however, little is known about the expression and the subcellular distribution of Cx43 in undifferentiated stem cells or about the structural arrangement of channels. We, therefore, here investigate expression of Cx43 in undifferentiated human cord-blood-derived induced pluripotent stem cells (hCBiPS2). For this purpose, we carried out quantitative real-time PCR and immunohistochemistry. For analysis of Cx43 ultrastructure and protein assembly, we performed freeze-fracture replica immunogold labeling (FRIL). Cx43 expression was detected at mRNA and protein level in hCBIPS2 cells. For the first time, ultrastructural data are presented on gap junction morphology in induced pluripotent stem (iPS) cells from cord blood: Our FRIL and electron microscopical analysis revealed the occurrence of gap junction plaques in undifferentiated iPS cells. In addition, these gap junctions were shown to contain the gap junction protein Cx43.

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Abbreviations

iPSC:: Induced pluripotent stem cells
Cx43:: Connexin 43
FRIL:: Freeze-fracture replica immunogold labeling
hCBiPS2:: Human cord-blood-derived induced pluripotent stem cell clone 2
hESC:: Human embryonic stem cells

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Acknowledgments

The authors wish to thank Andrea Rabung and Alexander Grissmer for excellent technical assistance. They also thank Ann Soether for linguistic editing and Alina Mattheis for the schematic illustrations. The authors are very grateful to Prof. Dr. John E. Rash for his continuous support and expert advice on the manuscript. The authors acknowledge financial support by the German Research Foundation and the Saarland, who funded the freeze-fracture unit. This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

Conflict of interest

All authors declare that research was done without any potential conflict of interest. No competing financial interests exist.

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Affiliations

Department of Anatomy and Cell Biology, Saarland University, Kirrberger Straße, Building 61, 66421, Homburg/Saar, Germany
Anja Beckmann, Nadine Hainz, Thomas Tschernig & Carola Meier
Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiac, Thoracic, Transplant and Vascular Surgery, Hannover Medical School, 30625, Hannover, Germany
Madline Schubert, Alexandra Haase & Ulrich Martin
REBIRTH-Cluster of Excellence, Hannover Medical School, 30625, Hannover, Germany
Madline Schubert, Alexandra Haase & Ulrich Martin

Authors

Anja Beckmann
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Madline Schubert
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Nadine Hainz
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Alexandra Haase
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Ulrich Martin
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Thomas Tschernig
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Carola Meier
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Correspondence to Carola Meier.

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Beckmann, A., Schubert, M., Hainz, N. et al. Ultrastructural demonstration of Cx43 gap junctions in induced pluripotent stem cells from human cord blood. Histochem Cell Biol 146, 529–537 (2016). https://doi.org/10.1007/s00418-016-1469-9

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Accepted: 15 July 2016
Published: 25 July 2016
Issue Date: November 2016
DOI: https://doi.org/10.1007/s00418-016-1469-9

Keywords

iPSC
Cx43
Freeze-fracture replica immunogold labeling