Zahabiya H Chithiwala1, Hoi Chang Lee2, David L Hill3, Teru Jellerette-Nolan4, Rafael Fissore2, Daniel Grow4, Daniel A Dumesic5. 1. Department of Obstetrics and Gynecology, University of California Los Angeles, 10833 Le Conte Ave, Room 22-178 CHS, Los Angeles, CA, 90095, USA. 2. Department of Veterinary and Animal Sciences, University of Massachusetts, Amherst, MA, 01003, USA. 3. ART Reproductive Center, 450 North Roxbury Drive Suite 520, Beverly Hills, CA, 90210, USA. 4. Division of Reproductive Endocrinology, Department of Obstetrics and Gynecology, Baystate Medical Center, Western Campus of Tufts University School of Medicine, 280 Chestnut Street, Springfield, MA, 01199, USA. 5. Department of Obstetrics and Gynecology, University of California Los Angeles, 10833 Le Conte Ave, Room 22-178 CHS, Los Angeles, CA, 90095, USA. danieldumesic@aol.com.
Abstract
PURPOSE: The purpose of this study is to describe impaired oocyte fertilization from phospholipase C-zeta (PLC-ζ) deficiency in normal-appearing sperm that was successfully treated using calcium (Ca(2+)) ionophore with intracytoplasmic sperm injection (ICSI) of oocytes matured in vitro. METHODS: An infertile couple undergoing in vitro fertilization (IVF) experienced failed oocyte fertilization following ICSI with normal-appearing sperm. A semen sample collected from the patient was used to assess the expression of sperm PLC- ζ protein by Western blot analysis and immunofluorescence and PLC-ζ bioactivity by an in vitro model of Ca(2+) release. A second IVF cycle was performed using Ca(2+) ionophore with ICSI to enhance Ca(2+)-induced oocyte activation of oocytes matured in vitro. RESULTS: Sperm PLC-ζ protein deficiency was demonstrated by Western blot analysis and immunofluorescence and confirmed by reduced PLC-ζ bioactivity using an in vitro model of Ca(2+) release. Nevertheless, with this sperm and supplementation of Ca(2+) ionophore following ICSI, fertilization of four of six oocytes matured in vitro was obtained. In addition, four embryos underwent cleavage and two of them reached the blastocyst stage. Transfer of these blastocysts into the uterus led to a single pregnancy and live birth. CONCLUSIONS: Deficiency of PLC-ζ in normal-appearing human sperm is associated with impaired Ca(2+)-dependent oocyte activation during ICSI. Under this condition, use of Ca(2+) ionophore following ICSI of oocytes matured in vitro improves embryo developmental competence, possibly through the activation of Ca(2+)-dependent mechanisms governing fertilization and preimplantation embryogenesis.
PURPOSE: The purpose of this study is to describe impaired oocyte fertilization from phospholipase C-zeta (PLC-ζ) deficiency in normal-appearing sperm that was successfully treated using calcium (Ca(2+)) ionophore with intracytoplasmic sperm injection (ICSI) of oocytes matured in vitro. METHODS: An infertile couple undergoing in vitro fertilization (IVF) experienced failed oocyte fertilization following ICSI with normal-appearing sperm. A semen sample collected from the patient was used to assess the expression of sperm PLC- ζ protein by Western blot analysis and immunofluorescence and PLC-ζ bioactivity by an in vitro model of Ca(2+) release. A second IVF cycle was performed using Ca(2+) ionophore with ICSI to enhance Ca(2+)-induced oocyte activation of oocytes matured in vitro. RESULTS: Sperm PLC-ζ protein deficiency was demonstrated by Western blot analysis and immunofluorescence and confirmed by reduced PLC-ζ bioactivity using an in vitro model of Ca(2+) release. Nevertheless, with this sperm and supplementation of Ca(2+) ionophore following ICSI, fertilization of four of six oocytes matured in vitro was obtained. In addition, four embryos underwent cleavage and two of them reached the blastocyst stage. Transfer of these blastocysts into the uterus led to a single pregnancy and live birth. CONCLUSIONS: Deficiency of PLC-ζ in normal-appearing human sperm is associated with impaired Ca(2+)-dependent oocyte activation during ICSI. Under this condition, use of Ca(2+) ionophore following ICSI of oocytes matured in vitro improves embryo developmental competence, possibly through the activation of Ca(2+)-dependent mechanisms governing fertilization and preimplantation embryogenesis.
Authors: S L Taylor; S Y Yoon; M S Morshedi; D R Lacey; T Jellerette; R A Fissore; S Oehninger Journal: Reprod Biomed Online Date: 2009-12-28 Impact factor: 3.828
Authors: Katerina A Turner; Amarnath Rambhatla; Samantha Schon; Ashok Agarwal; Stephen A Krawetz; James M Dupree; Tomer Avidor-Reiss Journal: Cells Date: 2020-04-16 Impact factor: 6.600