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Proteomic characterization of macro-, micro- and nano-extracellular vesicles derived from the same first trimester placenta: relevance for feto-maternal communication
University of Auckland, New Zealand.
University of Otago, New Zealand.
University of Auckland, New Zealand.
Linköping University, Department of Medical and Health Sciences. Linköping University, Faculty of Medicine and Health Sciences. University of Auckland, New Zealand.
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2016 (English)In: Human Reproduction, ISSN 0268-1161, E-ISSN 1460-2350, Vol. 31, no 4, 687-699 p.Article in journal (Refereed) PublishedText
Abstract [en]

STUDY QUESTION: What proteins are carried by extracellular vesicles (EVs) released from normal first trimester placentae? SUMMARY ANSWER: One thousand five hundred and eighty-five, 1656 and 1476 proteins were characterized in macro-, micro-and nano-vesicles, respectively, from first trimester placentae, with all EV fractions being enriched for proteins involved in vesicle transport and inflammation. WHAT IS KNOWN ALREADY: Placental EVs are being increasingly recognized as important mediators of both healthy and pathological pregnancies. However, current research has focused on detecting changes in specific proteins in particular fractions of vesicles during disease. This is the first study to investigate the full proteome of different-sized fractions of EVs from the same first trimester placenta and highlights the differences/similarities between the vesicle fractions. STUDY DESIGN, SIZE, DURATION: A well-established ex vivo placental explant culture model was used to generate macro-, micro-and nano-vesicles from 56 first trimester placentae. Vesicle fractions were collected by differential ultracentrifugation, quantified and characterized. PARTICIPANTS/MATERIALS, SETTING, METHODS: Placental macro-, micro-and nano-vesicles were characterized by microscopy, dynamic light scattering and nanoparticle tracking analysis. The proteome of each EV fraction was interrogated using liquid chromatography-coupled tandem mass spectrometry. Results were validated by semi-quantitative western blotting. MAIN RESULTS AND THE ROLE OF CHANCE: A total of 1585, 1656 and 1476 proteins were identified in macro-, micro-and nanovesicles, respectively. One thousand one hundred and twenty-five proteins were shared between all three fractions while up to 223 proteins were unique to each fraction. Gene Ontology pathway analysis showed an enrichment of proteins involved in vesicle transport and inflammation in all three fractions of EVs. The expression levels of proteins involved in internalization of vesicles (annexin V, calreticulin, CD31, CD47), the complement pathway [C3, decay-accelerating factor (DAF), membrane cofactor protein (MCP), protectin] and minor histocompatibility antigens [ATP-dependent RNA helicase (DDX3), ribosomal protein S4 (RPS4)] were different between different-sized EVs. LIMITATIONS, REASONS FOR CAUTION: This study is largely hypothesis-generating in nature. It is important to validate these findings using EVs isolated from maternal plasma and the function of the different EV fractions would need further investigation. WIDER IMPLICATIONS OF THE FINDINGS: Our results support the concept that various EV factions can interact with different maternal cells and have unique effects to mediate feto-maternal communication during early pregnancy. This study also provides a list of candidate proteins, which may inform the identification of robust markers that can be used to isolate placental vesicles from the maternal blood in the future.

Place, publisher, year, edition, pages
Oxford University Press, 2016. Vol. 31, no 4, 687-699 p.
Keyword [en]
syncytial nuclear aggregates; syncytial knots; extracellular vesicle; microparticle; exosome; trophoblast deportation; trophoblastic debris
National Category
Basic Medicine
Identifiers
URN: urn:nbn:se:liu:diva-127779DOI: 10.1093/humrep/dew004ISI: 000374228200002PubMedID: 26839151OAI: oai:DiVA.org:liu-127779DiVA: diva2:927620
Note

Funding Agencies|University of Auckland Health Research Doctoral Scholarship; Freemasons Postgraduate Scholarship; School of Medicine Performance-based research fund (PBRF) grant

Available from: 2016-05-12 Created: 2016-05-12 Last updated: 2016-05-12

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Johansson, Caroline L.
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Department of Medical and Health SciencesFaculty of Medicine and Health Sciences
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