Abstract
Differentiation of human placental villous trophoblast includes syncytial fusion of cytotrophoblast forming syncytiotrophoblast. Early stages of the apoptosis cascade were described to be involved in this differentiation process. We investigated the role of the initiator caspase 8 in syncytial fusion in vitro, cultivating placental villous explants with or without caspase 8 antisense oligonucleotides or peptide inhibitors for up to 120 h. Trophoblast fusion and differentiation were assessed by confocal microscopy, immunohistochemistry and Western blot analysis. Culture with caspase 8 antisense oligonucleotides or peptide inhibitors reduced the fusion of cytotrophoblast with the syncytiotrophoblast, and resulted in multilayered cytotrophoblast. Caspase 8 expression was suppressed by antisense oligonucleotides and caspase 8 activities were reduced by peptide inhibitors. The organic anion-transporter hOAT-4 normally expressed in the cytotrophoblast and transferred into the syncytiotrophoblast by syncytial fusion was retained in the cytotrophoblast due to lack of fusion. We conclude that expression and activity of caspase 8 is a prerequisite for differentiation and syncytial fusion of cytotrophoblast cells.
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Abbreviations
- ADAM:
-
a disintegrin and a metalloproteinase domain
- C3:
-
caspase 3
- C8:
-
caspase 8
- CT:
-
cytotrophoblast
- DEVD:
-
amino acids aspartic acid–glutamic acid–valine–aspartic acid
- DISC:
-
death-inducing signalling complex
- FADD:
-
Fas-associated death domain
- HIV-Env:
-
human immunodeficiency virus-envelope protein
- hOAT-4:
-
human organic anion transporter 4
- IETD:
-
amino acids isoleucine–glutamic acid–threonine–aspartic acid
- PCNA:
-
proliferative cell nuclear antigen
- PS:
-
phosphatidylserine
- ST:
-
syncytiotrophoblast
- TRADD:
-
TNF receptor-associated death domain
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Acknowledgements
The excellent technical assistance of Uta Zahn is gratefully acknowledged. PK and BH are funded by the Rockefeller Foundation, Grants No. RF96020#76 and RF99021#114.
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Black, S., Kadyrov, M., Kaufmann, P. et al. Syncytial fusion of human trophoblast depends on caspase 8. Cell Death Differ 11, 90–98 (2004). https://doi.org/10.1038/sj.cdd.4401307
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DOI: https://doi.org/10.1038/sj.cdd.4401307
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