Phosphatidylserine efflux and intercellular fusion in a BeWo model of human villous cytotrophoblast

M. Das, B. Xu, L. Lin, S. Chakrabarti, Vijay Shivaswamy, N. S. Rote

Research output: Contribution to journalArticle

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Abstract

Phosphatidylserine (PS) efflux characterizes cytotrophoblast apoptosis and differentiation. To evaluate whether PS externalization and intercellular fusion were secondary to apoptosis, BeWo cells were induced to differentiate by forskolin or undergo apoptosis by staurosporine. PS externalization was measured by FITC-annexin V binding, and intercellular fusion was quantified by counting nuclei in syncytial cells. During forskolin treatment, vanadate decreased PS efflux by 78.0 per cent from 68.0 [5.3] (mean [SD] to 15.0 [8.8] Lum ( × 10 3 ) (P< 0.001), whereas Z-VAD-fmk had no effect (66.5 [7.3]). Vanadate decreased intercellular fusion from 78.1 per cent [4.1] fusion in uninhibited cultures to 23.4 per cent [2.5], compared with 10.0 per cent [1.7] in media alone. Z-VAD-fmk did not affect fusion (80.4 per cent [6.8]). Staurosporine induced PS efflux was not affected by vanadate (69.6 [5.5] Lum × 10 3 ), but was inhibited 87.8 per cent by Z-VAD-fmk; from 71.5 [6.2] to 8.7 [3.6] Lum ( × 10 3 ) (P<0.001). Apoptosis was measured by the TUNEL and COMET assays, lamin B fragmentation, activation of procaspase 3, mitochondrial membrane potential, and release of mitochondrial cytochrome c and apoptosis inducing factor. There was no indication of apoptosis associated with differentiation. Thus, PS efflux and intercellular fusion occurred through a vanadate-sensitive mechanism that was independent of apoptosis.

Original languageEnglish (US)
Pages (from-to)396-407
Number of pages12
JournalPlacenta
Volume25
Issue number5
DOIs
StatePublished - Jan 1 2004

Fingerprint

Phosphatidylserines
Trophoblasts
Vanadates
Apoptosis
Staurosporine
Colforsin
Lamin Type B
Apoptosis Inducing Factor
Fluorescein-5-isothiocyanate
Mitochondrial Membrane Potential
Annexin A5
In Situ Nick-End Labeling
Cytochromes c
Caspase 3

ASJC Scopus subject areas

  • Reproductive Medicine
  • Obstetrics and Gynecology
  • Developmental Biology

Cite this

Phosphatidylserine efflux and intercellular fusion in a BeWo model of human villous cytotrophoblast. / Das, M.; Xu, B.; Lin, L.; Chakrabarti, S.; Shivaswamy, Vijay; Rote, N. S.

In: Placenta, Vol. 25, No. 5, 01.01.2004, p. 396-407.

Research output: Contribution to journalArticle

Das, M. ; Xu, B. ; Lin, L. ; Chakrabarti, S. ; Shivaswamy, Vijay ; Rote, N. S. / Phosphatidylserine efflux and intercellular fusion in a BeWo model of human villous cytotrophoblast. In: Placenta. 2004 ; Vol. 25, No. 5. pp. 396-407.
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