title: Single blastomere removal from murine embryos is associated with activation of matrix metalloproteinases and Janus kinase/signal transducers and activators of transcription pathways of placental inflammation.
abstract: Single blastomere removal from cleavage-stage embryos, a common procedure used in conjunction with preimplantation genetic diagnosis (PGD), may affect reproductive outcomes. We hypothesized that negative pregnancy outcomes associated with PGD may be due to impairment of placental signaling pathways. The goal of this study was to determine the molecular mechanisms through which placental signaling is deregulated by blastomere removal. Four-cell stage murine embryos produced by in vitro fertilization were subjected to removal of a single blastomere (biopsied) or to the same manipulations without the blastomere removal (controls). Placental tissues from term (18.5 day) pregnancies obtained after embryo transfer were tested for levels of nitrosative species, interleukin 6, signal transducers and activators of transcription (STAT) 1 and 3, suppressors of cytokine signaling (SOCS) 1, 2 and 3 and matrix metalloproteinases (MMP) 1, 2, 3 and 9. Significant increases in nitrosative stress (P < 0.05), phosphorylative activation of STAT1 (P < 0.05) but not STAT3, lower levels of the inhibitors SOCS2 (P < 0.01) and SOCS3 (P < 0.001) and activation of MMP9 (P < 0.001) were observed in placentas derived from biopsied embryos, compared with controls. Such effects could contribute to greater levels of premature membrane rupture, incorrect parturition, preterm birth and intrauterine growth restriction associated with PGD. This work has determined signaling mechanisms that may be responsible for blastomere removal effects on placental function, with the potential to become targets for improving obstetric and neonatal outcomes in assisted reproduction.
authors.affiliation: Department of Tropical Medicine, Medical Microbiology and Pharmacology, John A. Burns School of Medicine, University of Hawaii, 651 Ilalo Street, Honolulu, HI 96813, USA Graduate Program in Cellular and Molecular Biology, John A. Burns School of Medicine, University of Hawaii, 651 Ilalo Street, Honolulu, HI 96813, USA.
authors.forename: Brittany L M
authors.initials: BL
authors.lastname: Sato
authors.affiliation: Institute for Biogenesis Research, John A. Burns School of Medicine, University of Hawaii, 1960 East-West Rd, Honolulu, HI 96822, USA.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Sugawara
authors.affiliation: Institute for Biogenesis Research, John A. Burns School of Medicine, University of Hawaii, 1960 East-West Rd, Honolulu, HI 96822, USA.
authors.forename: Monika A
authors.initials: MA
authors.lastname: Ward
authors.affiliation: Department of Tropical Medicine, Medical Microbiology and Pharmacology, John A. Burns School of Medicine, University of Hawaii, 651 Ilalo Street, Honolulu, HI 96813, USA Institute for Biogenesis Research, John A. Burns School of Medicine, University of Hawaii, 1960 East-West Rd, Honolulu, HI 96822, USA Faculty of Pharmaceutical Sciences, University of British Columbia, 2405 Wesbrook Mall, Vancouver, BC, Canada V6T 1Z3 abby.collier@ubc.ca.
authors.forename: Abby C
authors.initials: AC
authors.lastname: Collier
mesh_terms: D000818:Animals; D001706:Biopsy; D001757:Blastomeres / Q000201:enzymology* / Q000276:immunology; D002970:Cleavage Stage, Ovum / Q000201:enzymology* / Q000276:immunology; D046149:Embryo Culture Techniques; D004624:Embryo Transfer; D004789:Enzyme Activation; D005260:Female; D005307:Fertilization in Vitro; D005865:Gestational Age; D007249:Inflammation / Q000201:enzymology / Q000209:etiology* / Q000276:immunology; D018836:Inflammation Mediators / Q000378:metabolism; D053612:Janus Kinases / Q000378:metabolism*; D020782:Matrix Metalloproteinases / Q000378:metabolism*; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D010766:Phosphorylation; D010920:Placenta / Q000201:enzymology* / Q000276:immunology; D011247:Pregnancy; D019836:Preimplantation Diagnosis / Q000009:adverse effects*; D012307:Risk Factors; D050791:STAT Transcription Factors / Q000378:metabolism*; D015398:Signal Transduction; D017392:Thiobarbituric Acid Reactive Substances / Q000378:metabolism
chemical_list: D018836:Inflammation Mediators; D050791:STAT Transcription Factors; D017392:Thiobarbituric Acid Reactive Substances; D053612:Janus Kinases; D020782:Matrix Metalloproteinases
keywords: assisted reproduction technology; in vitro fertilization; suppressors of cytokine signaling