title: Studies of the dynamics of nuclear clustering in human syncytiotrophoblast.
abstract: Syncytial nuclear aggregates (SNAs), clusters of nuclei in the syncytiotrophoblast of the human placenta, are increased as gestation advances and in pregnancy pathologies. The origins of increased SNAs are unclear; however, a better appreciation of the mechanism may give insight into placental ageing and factors underpinning dysfunction. We developed three models to investigate whether SNA formation results from a dynamic process of nuclear movement and to generate alternative hypotheses. SNA count and size were measured in placental explants cultured over 16 days and particles released into culture medium were quantified. Primary trophoblasts were cultured for 6 days. Explants and trophoblasts were cultured with and without cytoskeletal inhibitors. An in silico model was developed to examine the effects of modulating nuclear behaviour on clustering. In explants, neither median SNA number (108 SNA/mm(2) villous area) nor size (283 μm(2)) changed over time. Subcellular particles from conditioned culture medium showed a wide range of sizes that overlapped with those of SNAs. Nuclei in primary trophoblasts did not change position relative to other nuclei; apparent movement was associated with positional changes of the syncytial cell membrane. In both models, SNAs and nuclear clusters were stable despite pharmacological disruption of cytoskeletal activity. In silico, increased nuclear movement, adhesiveness and sites of cytotrophoblast fusion were related to nuclear clustering. The prominence of SNAs in pregnancy disorders may not result from an active process involving cytoskeleton-mediated rearrangement of syncytial nuclei. Further insights into the mechanism(s) of SNA formation will aid understanding of their increased presence in pregnancy pathologies.
authors.affiliation: Maternal and Fetal Health Research CentreInstitute of Human Development, School of Medicine, University of Manchester, Manchester, UK St Mary's HospitalCentral Manchester University Hospitals NHS Foundation Trust, Manchester Academic Health Science Centre, Manchester, UK s.j.calvert87@gmail.com.
authors.forename: S J
authors.initials: SJ
authors.lastname: Calvert
authors.affiliation: Department of Obstetrics and GynecologyWashington University School of Medicine, St Louis, Missouri, USA.
authors.forename: M S
authors.initials: MS
authors.lastname: Longtine
authors.affiliation: School of MathematicsAlan Turing Building, University of Manchester, Manchester, UK.
authors.forename: S
authors.initials: S
authors.lastname: Cotter
authors.affiliation: Maternal and Fetal Health Research CentreInstitute of Human Development, School of Medicine, University of Manchester, Manchester, UK St Mary's HospitalCentral Manchester University Hospitals NHS Foundation Trust, Manchester Academic Health Science Centre, Manchester, UK.
authors.forename: C J P
authors.initials: CJ
authors.lastname: Jones
authors.affiliation: Maternal and Fetal Health Research CentreInstitute of Human Development, School of Medicine, University of Manchester, Manchester, UK St Mary's HospitalCentral Manchester University Hospitals NHS Foundation Trust, Manchester Academic Health Science Centre, Manchester, UK.
authors.forename: C P
authors.initials: CP
authors.lastname: Sibley
authors.affiliation: Maternal and Fetal Health Research CentreInstitute of Human Development, School of Medicine, University of Manchester, Manchester, UK St Mary's HospitalCentral Manchester University Hospitals NHS Foundation Trust, Manchester Academic Health Science Centre, Manchester, UK.
authors.forename: J D
authors.initials: JD
authors.lastname: Aplin
authors.affiliation: Department of Obstetrics and GynecologyWashington University School of Medicine, St Louis, Missouri, USA.
authors.forename: D M
authors.initials: DM
authors.lastname: Nelson
authors.affiliation: Maternal and Fetal Health Research CentreInstitute of Human Development, School of Medicine, University of Manchester, Manchester, UK St Mary's HospitalCentral Manchester University Hospitals NHS Foundation Trust, Manchester Academic Health Science Centre, Manchester, UK.
authors.forename: A E P
authors.initials: AE
authors.lastname: Heazell
mesh_terms: D002462:Cell Membrane / Q000648:ultrastructure*; D002467:Cell Nucleus / Q000648:ultrastructure*; D003599:Cytoskeleton / Q000648:ultrastructure*; D005260:Female; D005455:Fluorescent Antibody Technique; D006801:Humans; D010920:Placenta / Q000648:ultrastructure*; D011247:Pregnancy; D059008:Time-Lapse Imaging; D014327:Trophoblasts / Q000648:ultrastructure*