Fertilization and Cleavage Axes Differ In Primates Conceived By Conventional (IVF) Versus Intracytoplasmic Sperm Injection (ICSI)

Abstract With nearly ten million babies conceived globally, using assisted reproductive technologies, fundamental questions remain; e.g., How do the sperm and egg DNA unite? Does ICSI have consequences that IVF does not? Here, pronuclear and mitotic events in nonhuman primate zygotes leading to the...

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Main Authors: Calvin R. Simerly, Diana Takahashi, Ethan Jacoby, Carlos Castro, Carrie Hartnett, Laura Hewitson, Christopher Navara, Gerald Schatten
Format: Article
Language:English
Published: Nature Publishing Group 2019-10-01
Series:Scientific Reports
Online Access:https://doi.org/10.1038/s41598-019-51815-4
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spelling doaj-936b0f4f351b4142b33f5458069f4be02020-12-08T07:49:33ZengNature Publishing GroupScientific Reports2045-23222019-10-019111410.1038/s41598-019-51815-4Fertilization and Cleavage Axes Differ In Primates Conceived By Conventional (IVF) Versus Intracytoplasmic Sperm Injection (ICSI)Calvin R. Simerly0Diana Takahashi1Ethan Jacoby2Carlos Castro3Carrie Hartnett4Laura Hewitson5Christopher Navara6Gerald Schatten7Pittsburgh Development Center, Division of Developmental & Regenerative Medicine, and Obstetrics-Gynecology-Reproductive Sciences, Cell Biology, and Bioengineering, University of Pittsburgh Cancer Institute, University of Pittsburgh School of MedicineDivision of Cardiometabolic Health, Oregon National Primate Research Center, Oregon Health & Science University, BeavertonCCRM Houston Main Center Memorial CityPittsburgh Development Center, Division of Developmental & Regenerative Medicine, and Obstetrics-Gynecology-Reproductive Sciences, Cell Biology, and Bioengineering, University of Pittsburgh Cancer Institute, University of Pittsburgh School of MedicinePittsburgh Development Center, Division of Developmental & Regenerative Medicine, and Obstetrics-Gynecology-Reproductive Sciences, Cell Biology, and Bioengineering, University of Pittsburgh Cancer Institute, University of Pittsburgh School of MedicineThe Johnson Center for Child Health and DevelopmentDepartment of Biology, South Texas Center for Emerging Infectious Disease, University of Texas at San AntonioPittsburgh Development Center, Division of Developmental & Regenerative Medicine, and Obstetrics-Gynecology-Reproductive Sciences, Cell Biology, and Bioengineering, University of Pittsburgh Cancer Institute, University of Pittsburgh School of MedicineAbstract With nearly ten million babies conceived globally, using assisted reproductive technologies, fundamental questions remain; e.g., How do the sperm and egg DNA unite? Does ICSI have consequences that IVF does not? Here, pronuclear and mitotic events in nonhuman primate zygotes leading to the establishment of polarity are investigated by multidimensional time-lapse video microscopy and immunocytochemistry. Multiplane videos after ICSI show atypical sperm head displacement beneath the oocyte cortex and eccentric para-tangential pronuclear alignment compared to IVF zygotes. Neither fertilization procedure generates incorporation cones. At first interphase, apposed pronuclei align obliquely to the animal-vegetal axis after ICSI, with asymmetric furrows assembling from the male pronucleus. Furrows form within 30° of the animal pole, but typically, not through the ICSI injection site. Membrane flow drives polar bodies and the ICSI site into the furrow. Mitotic spindle imaging suggests para-tangential pronuclear orientation, which initiates random spindle axes and minimal spindle:cortex interactions. Parthenogenetic pronuclei drift centripetally and assemble astral spindles lacking cortical interactions, leading to random furrows through the animal pole. Conversely, androgenotes display cortex-only pronuclear interactions mimicking ICSI. First cleavage axis determination in primates involves dynamic cortex-microtubule interactions among male pronuclei, centrosomal microtubules, and the animal pole, but not the ICSI site.https://doi.org/10.1038/s41598-019-51815-4
collection DOAJ
language English
format Article
sources DOAJ
author Calvin R. Simerly
Diana Takahashi
Ethan Jacoby
Carlos Castro
Carrie Hartnett
Laura Hewitson
Christopher Navara
Gerald Schatten
spellingShingle Calvin R. Simerly
Diana Takahashi
Ethan Jacoby
Carlos Castro
Carrie Hartnett
Laura Hewitson
Christopher Navara
Gerald Schatten
Fertilization and Cleavage Axes Differ In Primates Conceived By Conventional (IVF) Versus Intracytoplasmic Sperm Injection (ICSI)
Scientific Reports
author_facet Calvin R. Simerly
Diana Takahashi
Ethan Jacoby
Carlos Castro
Carrie Hartnett
Laura Hewitson
Christopher Navara
Gerald Schatten
author_sort Calvin R. Simerly
title Fertilization and Cleavage Axes Differ In Primates Conceived By Conventional (IVF) Versus Intracytoplasmic Sperm Injection (ICSI)
title_short Fertilization and Cleavage Axes Differ In Primates Conceived By Conventional (IVF) Versus Intracytoplasmic Sperm Injection (ICSI)
title_full Fertilization and Cleavage Axes Differ In Primates Conceived By Conventional (IVF) Versus Intracytoplasmic Sperm Injection (ICSI)
title_fullStr Fertilization and Cleavage Axes Differ In Primates Conceived By Conventional (IVF) Versus Intracytoplasmic Sperm Injection (ICSI)
title_full_unstemmed Fertilization and Cleavage Axes Differ In Primates Conceived By Conventional (IVF) Versus Intracytoplasmic Sperm Injection (ICSI)
title_sort fertilization and cleavage axes differ in primates conceived by conventional (ivf) versus intracytoplasmic sperm injection (icsi)
publisher Nature Publishing Group
series Scientific Reports
issn 2045-2322
publishDate 2019-10-01
description Abstract With nearly ten million babies conceived globally, using assisted reproductive technologies, fundamental questions remain; e.g., How do the sperm and egg DNA unite? Does ICSI have consequences that IVF does not? Here, pronuclear and mitotic events in nonhuman primate zygotes leading to the establishment of polarity are investigated by multidimensional time-lapse video microscopy and immunocytochemistry. Multiplane videos after ICSI show atypical sperm head displacement beneath the oocyte cortex and eccentric para-tangential pronuclear alignment compared to IVF zygotes. Neither fertilization procedure generates incorporation cones. At first interphase, apposed pronuclei align obliquely to the animal-vegetal axis after ICSI, with asymmetric furrows assembling from the male pronucleus. Furrows form within 30° of the animal pole, but typically, not through the ICSI injection site. Membrane flow drives polar bodies and the ICSI site into the furrow. Mitotic spindle imaging suggests para-tangential pronuclear orientation, which initiates random spindle axes and minimal spindle:cortex interactions. Parthenogenetic pronuclei drift centripetally and assemble astral spindles lacking cortical interactions, leading to random furrows through the animal pole. Conversely, androgenotes display cortex-only pronuclear interactions mimicking ICSI. First cleavage axis determination in primates involves dynamic cortex-microtubule interactions among male pronuclei, centrosomal microtubules, and the animal pole, but not the ICSI site.
url https://doi.org/10.1038/s41598-019-51815-4
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