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Glycosylated Siglec-6 expression in syncytiotrophoblast-derived extracellular vesicles from preeclampsia placentas

Abstract:

INTRODUCTION: Preeclampsia (PE) is associated with an exaggerated maternal systemic inflammatory response. Throughout gestation, the placenta releases extracellular vesicles through the syncytiotrophoblast layer (STB) into the maternal circulation and this is increased in PE. Expression of Siglec-6, a transmembrane receptor of molecular weight 50 KDa, is upregulated in PE placental tissue.

METHODS: Here we investigated respective abundance of Siglec-6 in PE -and normal pregnancy- (NP) derived placental lysates (PL) and syncytiotrophoblast-derived extracellular vesicles (STBEV). STBEV from PE and NP placentas were isolated through dual-lobe placental perfusion and serial ultracentrifugation. Siglec-6 was characterized by immunohistochemistry, immunoblotting, mass spectrometry (MS), and deglycosylation.

RESULTS: Immunoblotting revealed the expected Siglec-6 (50 KDa) band present in both PE and NP PL, however an additional heavier band was observed at 70 KDa only in PE PL, but not in NP. When interrogating STBEV we saw an absence of the expected 50 KDa band but the 70 KDa was present predominantly only in the PE STBEV. Deglycosylation of PL and STBEV from PE showed that the 70 KDa and the 50 KDa bands were reduced to 48 KDa, suggesting glycosylation. Both 48 KDa and 70 KDa bands were subjected to MS, confirming Siglec-6 expression in both.

DISCUSSION: Our data shows that the inability to detect Siglec-6 in circulation might be due to the placenta secreting STBEV carrying a modified glycosylated form of Siglec-6 with a 70 KDa molecular weight, significantly and uniquely upregulated in PE STBEV.

Publication status:
Published
Peer review status:
Peer reviewed

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Publisher copy:
10.1016/j.bbrc.2020.09.081

Authors

More by this author
Institution:
University of Oxford
Division:
MSD
Department:
Women's & Reproductive Health
Role:
Author
More by this author
Institution:
University of Oxford
Division:
MSD
Department:
Women's & Reproductive Health
Sub department:
Women's & Reproductive Health
Role:
Author
More by this author
Institution:
University of Oxford
Division:
MSD
Department:
Women's & Reproductive Health
Role:
Author
More by this author
Institution:
University of Oxford
Division:
MSD
Department:
Women's & Reproductive Health
Role:
Author
More by this author
Institution:
University of Oxford
Division:
MSD
Department:
Women's & Reproductive Health
Role:
Author


Publisher:
Elsevier
Journal:
Biochemical and Biophysical Research Communications More from this journal
Volume:
533
Issue:
4
Pages:
838-844
Publication date:
2020-09-27
Acceptance date:
2020-09-19
DOI:
EISSN:
1090-2104
ISSN:
0006-291X
Pmid:
32998819


Language:
English
Keywords:
Pubs id:
1136793
Local pid:
pubs:1136793
Deposit date:
2020-12-11
ARK identifier:

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